ODC-SCI Public Data Sets

Assessment of Spinal DNA Damage Following L1 Contusion Spinal Cord Injury in Female C57Bl/6 Mice

DOI:10.34945/F5CC8T

DATASET CITATION:

Scheijen E. E.M., Veeningen N., Duwé S., Ivanova A., Van Broeckhoven J., Hendrix S., Wilson III D. M. (2025) Assessment of Spinal DNA Damage Following L1 Contusion Spinal Cord Injury in Female C57Bl/6 Mice. Open Data Commons for Spinal Cord Injury. ODC-SCI:1278 http://dx.doi.org/10.34945/F5CC8T

ABSTRACT:

STUDY PURPOSE: Assessing the presence, timing, and spinal location of DNA damage following spinal cord injury.

DATA COLLECTED: 12-week-old female C57Bl/6 mice received a 75 kdyne contusion at spinal L1, were Sham-operated, or remained naive. At 1 hour post-injury (hpi), 1 day post-injury (dpi), 3 dpi, 7 dpi, and 28 dpi the mice were sacrificed (n = 6-10 per group). Histological analysis of the spinal cord was performed on longitudinal sections of the spinal cord. DNA damage was labeled using gamma-H2AX, neurons by NeuN labeling, Caspase-3 mediated apoptosis by Cleaved Caspase-3 labeling, and cell nuclei using DAPI. Imaging was performed on a Zeiss Axioscan using the 647, 488, and DAPI channels. Automatic analysis was performed using QuPath software. By thresholding, cells/neurons with gamma-H2AX foci (DNA-damaged cells), pan nuclear gamma-H2AX signal (cells with early cell death), and negative gamma-H2AX signal were identified and counted.

CONCLUSIONS: DNA damage accumulates rapidly (1 hour to 3 days post-injury) in the spinal cord following a severe SCI and the neuronal population is specifically affected.

PLX5622 Efficacy on Microglia Depletion and Its Impact on Functional Recovery Following Unilateral Dorsal Quadrant C4 Cervical Spinal Cord Injury in Female Lewis Rats

DOI:10.34945/F5MS4M

DATASET CITATION:

Cucarian J. Daniel., Rakheja R. Vikas., Scatterty K. R., Armstrong E., Storvold S., Nguyen A., Fouad K. (2025) PLX5622 Efficacy on Microglia Depletion and Its Impact on Functional Recovery Following Unilateral Dorsal Quadrant C4 Cervical Spinal Cord Injury in Female Lewis Rats. Open Data Commons for Spinal Cord Injury. ODC-SCI:1253 http://dx.doi.org/10.34945/F5MS4M

ABSTRACT:

STUDY PURPOSE: After spinal cord injury (SCI), microglia play a key role in the inflammatory response. Beyond its impact on tissue damage, inflammation has also been linked to adaptive and maladaptive plasticity. In this study, we aimed to evaluate the impact of depleting microglia following SCI in rats with unilateral incomplete cervical SCI using PLX5622. Initially, we aimed to evaluate the efficacy of PLX5622 in depleting microglia within the central nervous system of rats with SCI, followed by assessing the impact of microglial reduction on functionality in this model.

DATA COLLECTED: This experiment involved 14 age-matched adult female Lewis rats, 10–12 weeks old, divided equally into two groups (n=7 per group). Sensorimotor and behavioral assessments were performed before SCI induction, including the Open Field, Elevated Plus Maze, and Light-Dark Box tests. All rats received a C4 dorsolateral quadrant transection. Following a recovery period of 7 days, one group of rats was fed a diet containing PLX5622, a colony-stimulating factor 1 receptor inhibitor for microglia/macrophages, at 1200 ppm for 30 days. All behavioral tests were repeated during the PLX5622 treatment. At the end of the PLX administration, the rats were euthanized and perfused, and their brains, spinal cord, and liver tissues were harvested for further evaluation. Video analysis of the rats' behaviors was performed using customized programs developed in Python. To test the efficacy of PLX5622 on microglia depletion and morphology, tissue samples from the cingulate cortex, hypothalamus, and striatum, as well as the grey and white matter from the C1 segment and the lesion site C4, were processed. Additionally, the left liver lobe was collected to examine liver-resident macrophages (Kupffer cells) and evaluate the specificity of PLX5622-induced microglial depletion. Microglial and Kupffer cell quantification was performed using QuPath and ImageJ-Fiji. Morphological analysis of microglia was conducted with ImageJ-Fiji, and 3D microglial reconstructions in the regions of interest were performed using Imaris software. To assess the extent of SCI damage, the injured and spared areas within the spinal cord were measured using ImageJ-Fiji, along with the thickness of the scar tissue in the lesion core.

CONCLUSIONS: Our findings show that four weeks of PLX5622 treatment in female rats with incomplete cervical SCI resulted in a reduction of approximately half of the microglial population in the brain and spinal cord. While PLX5622 did not notably affect behavior, lesion size, or microglial morphology, it reduced scar tissue thickness at the lesion core, altered liver weight, and significantly decreased liver-resident macrophages.

An optogenetic model of hindlimb spasticity after complete T9-T10 spinal cord transection in male and female C57bl/6 mice

DOI:10.34945/F5H308

DATASET CITATION:

Goltash S., Khodr R., Bui T. V., Laliberte A. M. (2025) An optogenetic model of hindlimb spasticity after complete T9-T10 spinal cord transection in male and female C57bl/6 mice. Open Data Commons for Spinal Cord Injury. ODC-SCI:1276 http://dx.doi.org/10.34945/F5H308

ABSTRACT:

STUDY PURPOSE: The purpose of this study was to test the efficacy of optogenetic stimulation of cutaneous VGLUT2+ sensory afferents in eliciting spasticity-associated behaviours after spinal cord injury. A tri-hybrid transgenic mouse line was produced (Isl1-Cre : Vglut2-IRES2-FLPo-D : Ai80(RCFL-CatCh)-D) to express the CatCh channelrhodopsin variant with VGLUT2+ sensory afferents. These mice were then subjected to a T9-T10 complete spinal cord transection to induce spasticity. The primary goal of this work was to validate this mouse model for the study of hindlimb spasticity after spinal cord injury, where light could be used to simply and reliably elicit spasticity-associated behaviours.

DATA COLLECTED: EMG data from the tibialis anterior and gastrocnemius muscles of awake and freely behaving mice was collected in the weeks following a complete T9-T10 transection (2 wpi, 3 wpi, 4 wpi, 5 wpi, and 10 wpi : wpi=weeks post injury). Blue light was directed onto each of the paws of the mice through a transparent floored chamber. Each session was comprised of 9 light pulses directed to each hindpaw, as well as 2 control pulses directed away from the mouse (20 total sweeps). Amplitudes and latencies of the EMG responses were averaged across trials. Anatomical data examining the expression of markers of various sensory afferent subtypes were also collected to confirm the identities of the optogenetically stimulated sensory afferents. RNAScope in situ hybridization (VGLUT1, VGLUT2) and immunohistochemistry (TRPV1, CGRP, IB4, TDTOMATO) techniques were performed for these markers and cell counts were performed.

CONCLUSIONS: The results of these recordings demonstrated significant increases in the amplitude of EMG responses to the light stimulus from 2 wpi to 10 wpi, suggesting increased excitability of cutaneous sensorimotor pathways. Interestingly, this effect was significantly greater in the female cohort than in the males. Incidences of prolonged involuntary muscle contraction in response to the stimulus (fictive spasms) were also detected through EMG and visual observation during the testing period, supporting the presence of spasticity. As such, the optogenetic mouse model developed for this study appears to elicit spasticity-associated behaviours in SCI mice reliably and may be valuable for studying SCI-related limb spasticity mechanisms and therapeutic interventions

Pleiotrophin and Rehabilitative Training Following an Incomplete Cervical Dorsolateral Quadrant Spinal Cord Injury in Adult Female Lewis Rats

DOI:10.34945/F59K54

DATASET CITATION:

Ng C., Strayer A. J., Smith K., Lowe K., Gupta S. J., Yousuf M. S., Maguire A. D., Kerr B. J., Winship I. R., Fouad K., Fenrich K. K. (2025) Pleiotrophin and Rehabilitative Training Following an Incomplete Cervical Dorsolateral Quadrant Spinal Cord Injury in Adult Female Lewis Rats. Open Data Commons for Spinal Cord Injury. ODC-SCI:1234 http://dx.doi.org/10.34945/F59K54

ABSTRACT:

STUDY PURPOSE: To test the capabilities of pleiotrophin (PTN) to promote forelimb motor recovery after a cervical dorsal lateral quadrant (DLQ) partial transection spinal cord injury (SCI) in adult rats and to evaluate PTN as a potential treatment in the clinic.

DATA COLLECTED: Adult female Lewis rats (n = 24) were trained to perform single pellet reaching, grasping and retrieval (SPRGR) task prior to SCI to establish baseline performance and determine the preferred paw. Once trained, a C4 DLQ was performed ipsilateral to the preferred paw, using a custom-made scalpel blade. Immediately following SCI, PBS (1 µl; n = 12) or PTN (1.2 µg in 1 µl; n = 12) was injected into the ipsilesional intermediate gray matter (0.75 mm lateral of midline, 1.5 mm deep from surface), 2.5 mm caudal to the SCI site. Animals received SPRGR rehabilitative training starting a week after SCI. Training occurred 5 days/week for 10 min/day until the success rate of all rats plateaued (~5 weeks). The training was video recorded and analyzed in a blinded manner. Von Frey hair test was used to assess bilateral forepaw tactile sensitivity before and at 5 weeks post-SCI. Horizontal ladder task was used to assess limb function in a skilled locomotor task. All animals were video recorded on a horizontal ladder task prior to and 5 weeks post-SCI and analyzed in a blinded manner. Histological analysis of lesion severity was performed on transverse sections of the spinal cord showing the largest area of SCI damage. Sections were immune-stained with 0.5% cresyl violet solution and visualized under brightfield and phase-contrast microscopy.

CONCLUSIONS: PTN-treated rats performed worse on SPRGR tasks than control (PBS) rats following an incomplete cervical lesion. Both PTN and control rats had comparable lesion sizes. This approach requires further investigation to elucidate anatomical changes underlying worse functional outcomes in PTN-treated rats.

Cytosolic Phospholipase A2 in Infiltrating Monocyte-Derived Macrophages Does Not Impair Recovery After Thoracic Contusion Spinal Cord Injury in Female Mice

DOI:10.34945/F52307

DATASET CITATION:

Glaser E. P., Kopper T. J., Bailey W. M., Kumari R., Stewart A. S., Gensel J. C. (2024) Cytosolic Phospholipase A2 in Infiltrating Monocyte-Derived Macrophages Does Not Impair Recovery After Thoracic Contusion Spinal Cord Injury in Female Mice. Open Data Commons for Spinal Cord Injury. ODC-SCI:1237 http://dx.doi.org/10.34945/F52307

ABSTRACT:

STUDY PURPOSE: Our previous work suggests that cytosolic phospholipase A2 (cPLA2) plays a role in the proinflammatory potentiating effect of myelin on macrophages in vitro. The purpose of the current work was determine the role of macrophage cPLA2 expression on spinal cord injury (SCI) pathology and recovery. We first employed an in vitro model using bone marrow-derived macrophages isolated from cPLA2 KO mice. Next we optimized and generated cPLA2 KO bone marrow chimeras via hematopoietic stem cell transplantation with cPLA2 KO donors. Using our T9 contusion SCI model we examined locomotor recovery and tissue sparing after injury.

DATA COLLECTED: Bone marrow-derived macrophages isolated from cPLA2 KO or WT controls were treated with a combination of myelin and/or a proinflammatory stimulus (LPS+IFN-gamma). We collected in-vitro data examining reactive oxygen and nitrogen species production, neurotoxicity, and arginase activity after treatment. To first optimize our bone marrow chimerization technique we subjected C57BL/6J mice to different doses of total body irradiation: 8, 10.5, and 13 grays. We then transplanted hematopoietic stem cells from Actin-GFP donors and assessed chimerization efficiency 9 weeks after transplantation. Using flow cytometry, we quantified the percent of CD45+ leukocytes and CD45+/CD11b +/Ly6G - myeloid cells that were GFP positive. We then generated cPLA2 KO chimeras using cPLA2 KO or WT bone marrow donors and 10 grays of radiation. 8-12 weeks after chimerization female C57BL/6J mice (2 -4 months old at time of chimerization) were subjected to T9 65kdyn contusion SCI. To assess recovery after SCI in the cPLA2 KO bone marrow chimeras we employed the Basso Mouse Scale (BMS), the Catwalk XT gait analysis system, and the horizontal ladder test. We specifically quantified BMS score, percent of correct paw placements out of total steps, as well as regularity index, hind paw base of support, hind paw stride length, and hind paw print area. To determine tissue sparing after SCI in the cPLA2 KO bone marrow chimeras we employed a double stain with neurofilament axonal stain and eriochrome cyanine myelin stain. We used this technique to distinguish between lesion and intact spinal cord tissue. We quantified tissue sparing at the lesion epicenter as well as 300 um rostral and caudal to the lesion epicenter. Additionally, we quantified the intralesional axon density in the lesion core. To confirm chimerization we determined cPLA2 expression in circulating leukocytes at the study endpoint using RT-qPCR. Additionally, we determined cPLA2 expression at the lesion epicenter (T9) 7 days after injury using a nanostring gene array. The data from this gene array is included in a supplementary file.

CONCLUSIONS: After SCI, cPLA2 KO bone marrow recipient chimeras displayed similar locomotor recovery and tissue pathology compared to WT recipient chimera controls. These data suggest that although cPLA2 plays a critical role in myelin-mediated potentiation of proinflammatory macrophage activation in vitro, it may not contribute to secondary injury pathology in vivo after SCI.

Treating a C5 contusion injury in female rats with sigmoid protein and 4-methylumbelliferone in combination with single pellet grasping training

DOI:10.34945/F5M01J

DATASET CITATION:

Armstrong E. A., Nguyen A. T., Vavrek R., Raposo P. J.F., Scatterty K., Cucarian J. D., Sekandary M., Nelson B., Fenrich K. K., Fouad K. (2024) Treating a C5 contusion injury in female rats with sigmoid protein and 4-methylumbelliferone in combination with single pellet grasping training. Open Data Commons for Spinal Cord Injury. ODC-SCI:1195 http://dx.doi.org/10.34945/F5M01J

ABSTRACT:

STUDY PURPOSE: We tested the effect of 4-methylumbelliferone (4-MU), an inhibitor for hyaluronan (HA) synthesis and Intracellular Sigmoid Protein, a PTP sigma antagonist, in combination with single pellet grasping (SPG) training on neuroplasticity and functional recovery following cervical spinal cord injury in rats.

DATA COLLECTED: All rats (n=30) were trained on single pellet grasping (SPG) using semi-automated dispensers. Paw preference was determined within the first week of training. After a plateau was reached in both single pellet reaching attempts and success rate, all rats received unilateral cervical spinal contusions (based on paw preference) at C5 using the Infinite Horizon impactor. Rats were assigned to 2 groups based on the severity of the lesion, as determined by the cylinder test on day 3 post SCI. Four rats were excluded (n=2 failed to learn SPG, n=1 no discernible deficit, n=1 euthanised due to health issues). The Control group (n=13) consisted of animals who received 1 ml saline (s.c.) daily and nutella twice/day for 5 days/week for a total of 8 weeks. The 4MU+ISP group (n=13) received 500µg intracellular sigmoid protein daily (s.c.) and a total of 0.5g/Kg 4 methylumbelliferone (MU), 5 days a week in nutella. Treatments started at week 18 post SCI and continued through week 25 post SCI, for 8 weeks in total. SPG training started 18 weeks post injury (5x/week for 10 min each) for twelve weeks. Every 2 weeks during the training/treatment the rats were tested on the horizontal ladder. The treatments were stopped after 8 weeks and the rats were tested with cylinder, elevated plus maze, Irvine, Beatties and Bresnahan (IBB) Forelimb Scale, Forelimb Locomotor Score (FLS) and horizontal ladder tests. AAV9 tracering was administered bilaterally into the forelimb motor cortex to trace lesioned and unlesioned corticospinal tract (CST) tracts. The rats were euthanized 3 weeks after tracer injection. Livers were dissected and weighed, the brain and spinal cord cryoprotected for histologic examination. The spinal cord tissue damage was determined from GFAP immunostained serial horizontal section images by measuring the total area of damage. Analysis of collateral sprouting for both lesioned and unlesioned CST tracts, including the number of CST collaterals and the distance of collateral sprouting both rostral and caudal to the lesion from transverse sections.

CONCLUSIONS: Although there were no significant differences in the behavior post treatment there was a significant increase in CST sprouting in the treated animals.

Histological measurement of interstitial fluid flow and relationship to lesion volume after C4 hemicontusion in male and female rats

DOI:10.34945/F54016

DATASET CITATION:

Kwon H. Y., Cornelison C. (2024) Histological measurement of interstitial fluid flow and relationship to lesion volume after C4 hemicontusion in male and female rats. Open Data Commons for Spinal Cord Injury. ODC-SCI:1121 http://dx.doi.org/10.34945/F54016

ABSTRACT:

STUDY PURPOSE: To measure the effects of spinal cord injury on interstitial pressure and fluid flow and examine the potential role of interstitial fluid flow on lesion volume enlargement.

DATA COLLECTED: We collected interstitial pressure measurements at 1 hr, 3 days, and 7 days after contusion SCI in Sprague Dawley rats. We also used histological assessments to determine the distance of Evans Blue dye extravasation at days 3 and 7 after injury (proxy for interstitial fluid flow distance). Finally, we exogenously enhanced interstitial fluid flow using convection enhanced delivery at 7 days after injury and measured the resulting lesion volume compared to untreated controls.

CONCLUSIONS: Spinal interstitial pressure increases from -3 mmHg in the naive cord to a peak of 13 mmHg at 3 days post-injury (DPI) but relatively normalizes to 2 mmHg by 7 DPI. By quantifying vascular leakage of Evans Blue dye after a cervical hemi-contusion in rats, we confirm an increase in dye infiltration at 3 DPI compared to 7 DPI, suggestive of higher fluid velocities at the time of peak fluid pressure. Finally, exogenously enhancing interstitial flow significantly increases the resulting lesion volume.

Part I: A CLIMBER Meta-analysis, recovery time measured by behavioral outcome tests after contusive injuries on various spinal levels segments in rats and mice of both sexes from Literature-Extracted Data (LED)

DOI:10.34945/F5DG6D

DATASET CITATION:

Iorio E. G., Khanteymoori A., Fond K. A., Keller A. V., Maliga Davis L., Schwab J. M., Ferguson A. R., Torres-Espin A., Watzlawick R. (2024) Part I: A CLIMBER Meta-analysis, recovery time measured by behavioral outcome tests after contusive injuries on various spinal levels segments in rats and mice of both sexes from Literature-Extracted Data (LED). Open Data Commons for Spinal Cord Injury. ODC-SCI:1082 http://dx.doi.org/10.34945/F5DG6D

ABSTRACT:

STUDY PURPOSE: The purpose of the study was to analyze recovery after spinal cord injury through various different behavioral outcome tests. The study compared effect sizes from literature sourced (literature-extracted data (LED)) to the literatures’ corresponding publicly available raw data (individual animal data (IAD)). Random effect models and regression analyses were applied to evaluate predictors of neuro-conversion in LED versus IAD. Subgroup analyses were performed on animal sex, animal type, animal species, injury severity, injury segment and sample sizes. Publications with common injury models (contusive injuries) and standardized endpoints (open field assessments) were included in the meta-analyses. Studies that recorded open field assessments at 0-3 and 28-56 days past operation were included. This dataset includes the literature-extracted data (LED) (part 1) that was collected for the study. The code to replicate our study can be found on github (https://github.com/ucsf-ferguson-lab/climber_meta_analysis2024.git). This dataset corresponds with another dataset in ODC-SCI (10.34945/F5J59P) which contains raw data, individual animal data (IAD), that directly corresponds to the literature extracted data in this dataset.

DATA COLLECTED: The literature-extracted data (LED) contained in this dataset was pulled from numerical and graphical outcomes reported in published literature. This dataset includes data extracted from 7 different published articles. Unlike other datasets in odc-sci, each row represents an experimental group rather than an individual subject. The values are summarized for each experimental group. Each study from the published articles includes contusion injuries with various severities and different locations, which are indicated in this dataset. Different mice and rat species are included in the dataset with both sexes. Outcome scores at different days-post operation from BBB, BMS, Grooming and Forelimb Open Field tests are also included. The behavioral outcome scores over days post operation were used to calculate effect sizes.

CONCLUSIONS: The goal of the study was to compare effect sizes between the individual animal data (IAD) and the literature-extracted data (LED) using random effect models and regression analyses. Differences were observed in the number of experimental groups and animals per group, insufficient reporting of drop-out animals, and missing information on experimental details. Meta-analysis revealed differences in effect sizes between LED versus IAD. Publications with smaller sample sizes yielded larger effect sizes, while studies with larger sample sizes had smaller effects. Differences were observed between literature-extracted data (LED) and it’s corresponding individual animal data (IAD).

Investigating the temporal effects of spinal cord injury on cardiac function and structure in male rats with T3 complete transection and determining the primary cause of cardiac decline following spinal cord injury using male rats with T3 or L2 complete transection, or T2 severe contusion

DOI:10.34945/F59P4S

DATASET CITATION:

Fossey M. P.M., Balthazaar S. J.T., Squair J. W., Williams A. M., Poormasjedi-Meibod M., Nightingale T. E., Erskine E., Hayes B., Ahmadian M., Jackson G. S., Hunter D. V., Currie K. D., Tsang T. S.M., Walter M., Little J. P., Ramer M. S., Krassioukov A. V., West C. R. (2024) Investigating the temporal effects of spinal cord injury on cardiac function and structure in male rats with T3 complete transection and determining the primary cause of cardiac decline following spinal cord injury using male rats with T3 or L2 complete transection, or T2 severe contusion. Open Data Commons for Spinal Cord Injury. ODC-SCI:1059 http://dx.doi.org/10.34945/F59P4S

ABSTRACT:

STUDY PURPOSE: High-level spinal cord injury (SCI) alters cardiac function and causes cardiac atrophy in the chronic phase post-injury. How such events manifest over time post-injury and the primary cause of these cardiac changes were unknown. The first study (manuscript Part I) investigated the temporal changes in the heart on the acute-to-chronic continuum post-SCI. The remaining studies (manuscript Part II) investigated whether the primary cause of altered cardiac function post-SCI was the loss of sympathetic control to the heart.

DATA COLLECTED: In Part I a total of 66 male Wistar rats (10-11 weeks old at SCI) were randomly assigned to T3 complete transection SCI (performed with microscissors and suction) or a SHAM injury; rats were assessed for outcomes at different time-points along the acute-to-chronic continuum. Part I represents the complete data of 61 rats - the SCI was fatal in 4 rats (6.1% mortality rate, below UBC animal ethical board’s expectations of 10%), while one rat was excluded from data analysis due to poor health at termination. To study cardiac volumes, echocardiography was performed in the same rats over-time, at pre-surgery, 1 day, 2 days, 4 days, 6 days and 8 weeks post-SCI (n=6-10 per time-point). To study cardiac function, we performed left-ventricular (LV) catheterization in different rats at termination: 1 day, 3 days, 5 days, 7 days and 8 weeks post-SCI (n=6-9 per group). To estimate the temporal changes in sympathetic activity post-SCI, we collected blood at 1 day, 7 days and 8 weeks post-SCI to detect plasma norepinephrine (NE) levels via an ELISA (n=4-7 per group). To measure LV cardiomyocyte dimensions, we collected mid-ventricular cross-sectional discs of the heart for histology at 12 hours, 1 day, 3 days, 5 days, 7 days and 8 weeks post-SCI (n=5-9 per group). To investigate whether and when protein degradation pathways were at play post-SCI in the heart causing cardiac atrophy, we collected LV apex tissue at all acute time-points, 12 hours to 7 days, to perform qPCR (n=5 per group). In Part II, we performed three rodent studies. The first study aimed to determine whether the reduction in cardiac function post-SCI was neurally mediated. We performed in order: LV catheterization, a T3 complete transection SCI (same injury model as Part I) and a chemical ganglionic blockade (hexamethonium bromide, HEX; I.V. 20 mg/kg) in male rats (total n=7; n=4 Sprague Dawley 23 weeks old at SCI, n=3 Wistar 11-12 weeks old at SCI). We compared the cardiac functional outcomes nadir post-SCI and post-HEX. The second study (n=20 male Wistar rats, 10-11 weeks old at SCI), investigated the involvement of bulbospinal sympathetic control in reduced cardiac function post-SCI by comparing LV catheterization outcomes at 13 weeks following complete transections (same injury model as Part I) at the T3 (interrupted bulbospinal sympathetic control; n=6) and L2 level (intact bulbospinal sympathetic control; n=7), compared to SHAM controls (n=7). To further isolate the role of the bulbospinal sympathetic control in mediating these reductions, we treated T3 severely contused rats (400 kdyn, 5 second dwell time with Infinite Horizon impactor) with the neuroprotective agent minocycline or a vehicle control (total n=19; male Wistar rats, 10-12 weeks old at SCI). At eight weeks post-SCI, we performed LV catheterization to obtain cardiac functional outcomes (n=5 minocycline treated rats and n=6 vehicle treated controls), and histology to assess preservation of bulbospinal sympathetic fibers in the spinal cord (n=4 per treatment group). Encompassing all sub-studies, this dataset contains the data of 112 rats.

CONCLUSIONS: Part I - High-level SCI causes a rapid and sustained reduction in cardiac function and plasma NE levels, which precedes reductions in cardiac volumes and cardiomyocyte size. Markers pertaining to protein degradation are upregulated rapidly post-SCI. Part II - By performing a chemical ganglionic blockade (HEX) following a high-level SCI, we observed no further reduction in cardiac functional outcomes compared to the SCI alone demonstrating that the reduced cardiac function observed post-SCI is neurally mediated. When comparing the effects of complete transections at the T3 and L2 level on cardiac function, there was no reduction in cardiac function following L2-SCI as opposed to following T3-SCI; therefore, this suggests that interrupted bulbospinal sympathetic control plays a role in reducing cardiac function post-SCI. Minocycline treatment preserved both bulbospinal sympathetic fibers and cardiac function following a T3 severe contusion. In summary, Part II experiments implicate that the loss of bulbospinal sympathetic control over the heart is the primary cause of the reduction in cardiac function post-SCI. Our findings demonstrate the importance for early and/or neuroprotective interventions which focus on restoring sympathetic control to the cardiovascular system.

Ultrasound-based hemodynamic measurements (bolus kinetics) following a unilateral C5 contusion injury in female Long Evans rats

DOI:10.34945/F5259B

DATASET CITATION:

Harmon J. N., Hyde J. E., Jensen D. E., D'cessare E. C., Odarenko A. A., Bruce M. F., Khaing Z. Z. (2024) Ultrasound-based hemodynamic measurements (bolus kinetics) following a unilateral C5 contusion injury in female Long Evans rats. Open Data Commons for Spinal Cord Injury. ODC-SCI:889 http://dx.doi.org/10.34945/F5259B

ABSTRACT:

STUDY PURPOSE: Spinal cord injury (SCI) leads to significant perturbations in blood flow in and around the injury epicenter. Over time, these impairments can lead to ischemia and cell death. The purpose of this study is to use intravital contrast-enhanced ultrasound imaging to examine the evaluation of the unilateral cervical SCI using longitudinal imaging to quantify anatomical and hemodynamic changes in vivo through the entire spinal parenchyma.

DATA COLLECTED: Here, we used ultrasound imaging to visualize and quantify subacute injury expansion (through 72 hours post-injury) in a rodent cervical contusion model. We collected B-mode and contrast-enhanced ultrasound (CEUS) images at baseline, acutely after injury and at 4, 24 or 72 hours post injury (hpi) in three cohorts of animals. In this data set, spinal cord volume, hematoma volume, central sucal artery tortuorsity, area of perfusion deficit and direction of perfusion deficit expansion are included. We found a significant increase in hematoma size over the 72 hour window of examination (1.86±0.17-fold change from acute, p < 0.05), as well as in the areas of ischemic deficits observed at 24 hours post-injury (2.24±0.27-fold, p < 0.05). We also noted significant deformation of the central sulcal artery nearest to the lesion site. In conjunction, we observed significant hyperemia in all tissue regions surrounding the injury site, except for the ipsilesional gray matter.

CONCLUSIONS: This study highlights the unique advantages of longitudinal ultrasound imaging as a method for in-vivo monitoring of spinal injury evolution.

Ultrasound-based anatomical measurements following a unilateral C5 contusion injury in female Long Evans rats

DOI:10.34945/F5SP44

DATASET CITATION:

Harmon J. N., Hyde J. E., Jensen D. E., D’cessare E. C., Odarenko A. A., Bruce M. F., Khaing Z. Z. (2024) Ultrasound-based anatomical measurements following a unilateral C5 contusion injury in female Long Evans rats. Open Data Commons for Spinal Cord Injury. ODC-SCI:888 http://dx.doi.org/10.34945/F5SP44

ABSTRACT:

STUDY PURPOSE: Spinal cord injury (SCI) leads to significant perturbations in blood flow in and around the injury epicenter. Over time, these impairments can lead to ischemia and cell death. The purpose of this study is to use intravital contrast-enhanced ultrasound imaging to examine the evaluation of the unilateral cervical SCI using longitudinal imaging to quantify anatomical and hemodynamic changes in vivo through the entire spinal parenchyma.

DATA COLLECTED: Here, we used ultrasound imaging to visualize and quantify subacute injury expansion (through 72 hours post-injury) in a rodent cervical contusion model. We collected B-mode and contrast-enhanced ultrasound (CEUS) images at baseline, acutely after injury and at 4, 24 or 72 hours post injury (hpi) in three cohorts of animals. In this data set, spinal cord volume, hematoma volume, central sucal artery tortuorsity, area of perfusion deficit and direction of perfusion deficit expansion are included. We found a significant increase in hematoma size over the 72 hour window of examination (1.86±0.17-fold change from acute, p < 0.05), as well as in the areas of ischemic deficits observed at 24 hours post-injury (2.24±0.27-fold, p < 0.05). We also noted significant deformation of the central sulcal artery nearest to the lesion site. In conjunction, we observed significant hyperemia in all tissue regions surrounding the injury site, except for the ipsilesional gray matter.

CONCLUSIONS: This study highlights the unique advantages of longitudinal ultrasound imaging as a method for in-vivo monitoring of spinal injury evolution.

Neuropathic Pain, Depressive-like Behaviors and Myeloid Cell Polarization in LysM-eGFP Mice of Both Sexes Following a Moderate Unilateral Cervical SCI

DOI:10.34945/F5GC7C

DATASET CITATION:

Richards J. H., Freeman D. D., Detloff M. R. (2024) Neuropathic Pain, Depressive-like Behaviors and Myeloid Cell Polarization in LysM-eGFP Mice of Both Sexes Following a Moderate Unilateral Cervical SCI. Open Data Commons for Spinal Cord Injury. ODC-SCI:940 http://dx.doi.org/10.34945/F5GC7C

ABSTRACT:

STUDY PURPOSE: Neuropathic pain is a multifaceted, chronically debilitating condition that has sensory descriminative, affective and cognitive components. Following spinal cord injury (SCI), approximately 60% of individuals are diagnosed with neuropathic pain and comorbid mood disorders, while only ~21% of the general population will experience a mood disorder in their lifetime. These experiments aim to characterize development of pain- and mood-related behaviors and correlate them with the innate immune response post-SCI. While humanizing the rodent is impossible, the results from this study inform clinical literature to closely examine sex-differences reported in humans to better understand the underlying shared etiologies of pain and depressive-like behaviors following CNS trauma. We hypothesize that nociceptive and depressive-like dysregulation occurs after SCI and is associated with aberrant macrophage infiltration in segmental pain centers.

DATA COLLECTED: We completed moderate (40kDyn, 2sec dwell time) unilateral C5 spinal cord contusion on male and female LysM-eGFP reporter mice (age >6weeks) to visualize infiltrating macrophages. von Frey, Hargreaves, open-field and sucrose preference tests were conducted weekly, while the forced swim test and the mechanical conflict avoidance paradigm were only conducted at the terminal timepoint. At 6-weeks post-SCI, mice exhibit nociceptive and depressive-like dysfunction compared to naïve and sham groups. There were no differences between sexes, indicating that sex is not a contributing factor driving nociceptive or depressive-like behaviors after SCI. Group averages based on experimental group revealed that SCI caused persistent mechanical allodynia and thermal hyperalgesia with indications of both learned-helplessness and anhedonia at the terminal timepoint of the experiment. SCI mice displayed increased myeloid cell presence (infiltrating macrophages: LysM+/CD68+, resident macrophages: LysM-/CD68+, microglia: IBA1+) in the lesion epicenter, ipsilateral C7-8 dorsal horn and C7-8 DRGs as evidenced by eGFP, CD68, and Iba1 immunostaining when compared to naïve and sham mice. This was further confirmed by SCI-induced alterations in the expression of genes indicative of myeloid cell activation states and their associated secretome in the dorsal horn and DRGs.

CONCLUSIONS: Utilizing hierarchical cluster analysis, we classified mice based on endpoint nociceptive and depressive-like behavior scores. Approximately 59.3% of SCI mice clustered based on increased paw withdrawal threshold to mechanical stimuli and immobility time in the forced swim test. In conclusion, moderate unilateral cervical SCI caused the development of pain-related and depressive-like behaviors in a subset of mice and these behavioral changes are consistent with immune system activation in the segmental pain pathway.

Effects of IB4+ nociceptor ablation on motor and pain-like behaviors in a female Sprague Dawley rat model with a unilateral C5 contusion injury

DOI:10.34945/F5M599

DATASET CITATION:

Walker J. R., Espana R. A., Detloff M. Ryan. (2024) Effects of IB4+ nociceptor ablation on motor and pain-like behaviors in a female Sprague Dawley rat model with a unilateral C5 contusion injury. Open Data Commons for Spinal Cord Injury. ODC-SCI:786 http://dx.doi.org/10.34945/F5M599

ABSTRACT:

STUDY PURPOSE: The primary nociceptive C-fibers become hyperexcitable and sprout into the dorsal horn after cervical spinal cord injury. This study investigated whether this maladaptive nociceptor plasticity is causal to the stereotypical motor deficits seen in reaching and grasping.

DATA COLLECTED: An adult (~10weeks at start of experiment) female Sprague-Dawley rat (starting weight ~225g) unilateral C5 contusion model was used with unilateral C5 laminectomy controls. C7-8 DRGs received a single microinjection per DRG (2000nl/DRG at a rate of 200nl/min) of IB4 conjugated Saporin (1.2mg/ml) to ablate the IB4+ nociceptors or control, unconjugated saporin at the time of SCI. Rats were behaviorally tested for motor and pain like behaviors before and at 4 weeks post injury. Motor tests included the Vulintus Isometric Pull Task, Single Pellet Retrieval, Montoya Staircase, IBB Cereal Manipulation, and Cylinder test of Paw Preference. Pain-like behavior included Automated von Frey test for mechanical allodynia and Hargreaves' Test for thermal hyperalgesia. 10 sets of 25-micron sections of C7-8 cord and 10 sets of 20-micron sections of C7 and C8 DRGs underwent primary antibody and lectin stains nociceptor terminal arbors in the C7 and C8 dorsal horn and cell bodies in the dorsal root ganglia were identified via anti-CGRP and streptavidin conjugated-IB4 to quantify changes in sprouting and confirm cell ablation. Sprouting was quantified using ImageJ to determine the proportional area of immunofluorescent fibers. Cell ablation was confirmed by counting the number of IB4+ cells in the DRGs.

CONCLUSIONS: Ablation of IB4+ nociceptors at the time of spinal cord injury significantly improved the number of rats who were able to grasp a chocolate flavored food pellet compared to controls. Ablation did not yield significant improvements in other motor tests indicating a task specific mechanism of nociceptor involvement in forelimb motor control after SCI. Ablation of nociceptors in laminectomy controls did not have any adverse effects on motor control.

Impact of Anti-Inflammatory Medication on Task-Specific Training Efficacy and Functional Recovery After Unilateral Dorsal Quadrant C4 Cervical Spinal Cord Injury in Female Lewis Rats

DOI:10.34945/F57W2G

DATASET CITATION:

Cucarian J. Daniel., Raposo P., Nguyen A., Vavrek R., Torres-Espin A., Fouad K. (2023) Impact of Anti-Inflammatory Medication on Task-Specific Training Efficacy and Functional Recovery After Unilateral Dorsal Quadrant C4 Cervical Spinal Cord Injury in Female Lewis Rats. Open Data Commons for Spinal Cord Injury. ODC-SCI:955 http://dx.doi.org/10.34945/F57W2G

ABSTRACT:

STUDY PURPOSE: After spinal cord injury, inflammation is involved in secondary tissue damage. However, it may also promote neuroplasticity. We have shown earlier that promoting inflammation in a chronic setting in rats can promote the efficacy of rehabilitative training in a reaching task. Here we wanted to test whether the opposite is also true. Would common anti-inflammatory medications that could be given for any reason in later stages of a spinal lesion affect the efficacy of rehabilitative training in rats with unilateral incomplete cervical spinal cord injuries.

DATA COLLECTED: This experiment involved two experimental cohorts, with a total of fifty-three age-matched adult female Lewis rats (cohort 1: n=29, cohort 2: n=24). The rats underwent training in a single pellet grasping (SPG) task for 5 weeks before receiving a C4 dorsolateral quadrant transection. Afterwards, the rats were randomized into groups: In the first cohort, three groups were included, SCI only (n=10), SCI + Diphenhydramine (SCI+DPH; n=10), and SCI + Methylprednisolone (SCI + MP; n=9). In the second cohort, only the SCI and SCI+DPH groups were included, each with a n=12. One week after the spinal cord lesion, the rats received Diphenhydramine and Methylprednisolone at 20mg/kg and 30mg/kg, respectively in their drinking water for 4 weeks, in combination with eight weeks of SPG training (10min/day). Sensorimotor and behavioral assessments were carried out and video recorded, before the dorsolateral quadrant transection (baseline), as well as on a weekly basis following the lesion. These tests included the Horizontal Ladder, Open Field, Elevated Plus Maze, Light-dark box, Von Frey, and The Irvine, Beattie, and Bresnahan test. After the final day of testing, the rats were euthanized, perfused, and their spinal cord tissue was harvested. The cervical spinal cord tissue, including the lesion site, was cryosectioned at 25 microns and processed with Neurotrace staining. To quantify the extent of spinal cord injury, we measured the damaged and spared areas within the spinal cord using ImageJ-Fiji.

CONCLUSIONS: Despite the well-established anti-inflammatory properties of both Diphenhydramine and Methylprednisolone and their extended administration in this experiment, our findings indicate that they did not have significant effects on any of the outcome measures. These results indicate that although proinflammatory stimuli were reported to improve post spinal cord injury training efficacy, these commonly administered inflammatories (especially the anti histaminergic Diphenhydramine) do not affect forelimb grasping training efficacy.

Probiotic treatment in female Lewis rats following unilateral incomplete cervical (C5) contusion injury: Microbiome report

DOI:10.34945/F5CP4D

DATASET CITATION:

Raposo P., Nguyen A. T., Schmidt E. K.A., Torres Espin A., Fenrich K. K., Bennett D. J., Fouad K. (2023) Probiotic treatment in female Lewis rats following unilateral incomplete cervical (C5) contusion injury: Microbiome report. Open Data Commons for Spinal Cord Injury. ODC-SCI:688 http://dx.doi.org/10.34945/F5CP4D

ABSTRACT:

STUDY PURPOSE: The bidirectional communication between the gastrointestinal tract and the central nervous system appears to be linked to the intestinal microbiome. Research has shown that spinal cord injury (SCI) can disrupt the gut microbiome, leading to gut dysbiosis. These changes can have several negative impacts, such as exacerbated systemic inflammation and susceptibility to infection. Probiotics administered to mice with SCI have been shown to ameliorate gut dysbiosis, confer neuroprotection, and improve locomotor recovery. However, probiotics have also produced conflicting results, making potential claims regarding the application of probiotics as a therapeutic supplement ambiguous. This study aimed to investigate the effects of a commercially available probiotic on recovery in a rat model of cervical SCI.

DATA COLLECTED: Female Lewis rats (12 weeks and 180-220 g) received a C4-C5 laminectomy to expose the spinal cord. Using an Infinite Horizon impactor a 125 kdyn unilateral contusion was performed targeting the right side of the spinal C5 level (1.25 mm right of midline, 25 mm drop height at a 15-degree angle). Probiotic treatment was initiated immediately after SCI using VSL#3 containing 450 billion colony-forming units (CFU) of lyophilized bacteria/g, including eight different strains (Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus planatarum, and Streptococcus salivarus subspecies thermophilus). VSL#3 was administered daily via oral gavage for 7 days postinjury (DPI) at a dose of 5 billion CFU. Control group was gavaged with 0.5 ml of sterile filtered water. After 7 DPI, the 5 billion CFU of VSL#3 doses were adjusted to ensure equivalent consumption and administered ad libitum in drinking water for a total of 35 DPI. The overall gut microbiome was assessed prior to injury (baseline) and at 3, 7, 14, and 35 DPI by collecting fresh fecal matter from the rats during their dark cycle and submitted for 16S rRNA analysis. Dataset for Probiotic treatment for acute spinal cord injury: behavioural, histological and systemic cytokine profiles of female Lewis rats is in OSC-SCI#687

CONCLUSIONS: We could not detect any beneficial effects of the probiotic treatment. If anything it was detrimental as lesion size was slightly larger and the return of the gut microbiome to preinjury state was delayed by the treatment. Our results thus do not support the application of the utilized probiotic cocktail as a dietary supplement for the treatment of cervical SCI-induced gut dysbiosis and secondary complications.

C57BL/6J mice exposed to dim light-at-night following a T9 contusion spinal cord injury exhibit modest improvements in locomotor recovery accompanied by increased behaviors related to depression and mechanical neuropathic pain

DOI:10.34945/F5N30H

DATASET CITATION:

Aldrich J. C., Scheinfeld A. R., Lee S. E., Mahach K. M., Van de Veire B. C., Fonken L. K., Gaudet A. D. (2023) C57BL/6J mice exposed to dim light-at-night following a T9 contusion spinal cord injury exhibit modest improvements in locomotor recovery accompanied by increased behaviors related to depression and mechanical neuropathic pain. Open Data Commons for Spinal Cord Injury. ODC-SCI:956 http://dx.doi.org/10.34945/F5N30H

ABSTRACT:

STUDY PURPOSE: Nighttime disturbances are a common feature of extended hospital stays and likely lead to circadian disruption which, in turn, may exacerbate neuroinflammatory conditions. This study aimed to assess the effects of circadian disruption via exposure to dim light-at-night (dLAN) on locomotor recovery, tissue sparing, neuropathic pain sensitivity, and mood-related behaviors in mice following a T9 contusion spinal cord injury (SCI).

DATA COLLECTED: A more detailed methodology is available in the preprint: https://doi.org/10.1101/2023.09.15.557980. Briefly, male and female C57BL/6J mice were subjected to either moderate (65 kdyn) T9 contusion SCI or sham surgeries and subsequently housed under standard light-dark conditions (LD; 12 h light at 150 lux; 12 hr dark) or dLAN conditions (12 h light at 150 lux; 12 h dim light at 15 lux). The Basso mouse scale (BMS) was used to evaluate locomotor recovery at various timepoints from 1 to 35 days post-operation (dpo). Heat hyperalgesia and mechanical allodynia were assessed using the Hargreaves test and the simplified up-down (SUDO) von Frey method, respectively, at 6 or 7 dpo and weekly thereafter. In both tests, left and right hindpaw scores are averaged together and for Hargreaves each hindpaw was tested three times. Anxiety- and depressive-like behaviors were evaluated using the juvenile social exploration (JSE) test, sucrose preference test (SPT), and open-field test (OFT). JSE and OFT were performed prior to surgery and at 21 dpo while SPT was performed prior to surgery and at 14 and 29 dpo. For OFT and JSE, mouse behavior was tracked via Ethovision in an open area for 10 min followed by the introduction of a same-sex novel juvenile stimulus mouse (?4 weeks old) for 5 min. Distance traveled and percent time in the center 1/3rd of the arena for the first 5 min was recorded for OFT, while the amount of time spent interacting (e.g. sniffing, grooming, chasing, and fighting) with the juvenile was manually scored. For SPT, mice—housed alone or in sex-matched pairs, as noted in the dataset—were presented overnight with the choice of water or a 2% sucrose solution. Bottles containing water or sucrose were weighed before and after the test to determine the average amount consumed per mouse—i.e. in cases where two mice were housed together, the amount consumed was divided by two. Finally, neuroprotection was assessed in 35 dpo spinal cord cryosections by measuring the lesion area and total cross-sectional area—based on anti-GFAP immunofluorescence staining of the astrocytic border—at 0.2 mm intervals surrounding the injury epicenter. Percent tissue sparing was calculated thusly: (total area – lesion area) / total area * 100%.

CONCLUSIONS: Subjecting C57BL/6J mice to SCI and housing them in dLAN had minimal effects on locomotor and pain-related outcomes. SCI mice exposed to dLAN displayed slightly improved locomotor function at 28 dpo—and no other timepoint—with an average BMS score of 6.25 vs. 5.05 for LD housed animals. Post-SCI dLAN exposure resulted in worsened mechanical hypersensitivity at 13 dpo—filament threshold 6.5 dLAN vs. 7.75 LD—and no difference in heat hypersensitivity. The impact of dLAN on mood-related behaviors in SCI mice was overall limited . No significant differences were observed in the sucrose preference and open-field tests between the two housing conditions—however, interestingly, SCI-dLAN mice exhibited a ~23% reduction in time spent on juvenile social exploration compared to SCI-LD mice, suggesting a potential increase in depressive-like behavior. Finally, consistent with the mild locomotor effect, we saw no difference in tissue sparing or lesion size in 35 dpo spinal cord sections. Overall, our findings indicate that C57BL/6J mice exposed to dim light-at-night following spinal cord injury displayed modest effects on locomotor recovery, pain-like responses, and mood-related behaviors. These results emphasize the need for future investigations to delve deeper into the impact of circadian disruption on post-injury outcomes, exploring a range of manipulations such as sleep fragmentation, stress, and varying lighting conditions, either individually or in combination, to better simulate more complex clinical scenarios.

Lumbar (L2-L4) intermediate gray matter (laminae V-VIII) kainic acid-induced spinal cord injury in female Fisher 344 rats creates deficits in gross locomotion, coordination, balance and gait

DOI:10.34945/F5DK52

DATASET CITATION:

Kuehn N., Schwarz A., Beretta C. A., Schwarte Y., Schmitt F., Motsch M., Weidner N., Puttagunta R. (2023) Lumbar (L2-L4) intermediate gray matter (laminae V-VIII) kainic acid-induced spinal cord injury in female Fisher 344 rats creates deficits in gross locomotion, coordination, balance and gait. Open Data Commons for Spinal Cord Injury. ODC-SCI:938 http://dx.doi.org/10.34945/F5DK52

ABSTRACT:

STUDY PURPOSE: Spinal cord injury (SCI) often occurs at spinal enlargements where the gray matter is the largest for input and output from and to the limbs. Much SCI research focuses on long-distance axonal regeneration, neglecting the importance of replacing lost gray matter for functional recovery. Here we show that the loss of the intermediate gray matter (laminae V-VIII) of L2-L4 leads to behavioral deficits that cannot be compensated for over time. Thereby establishing a lumbar intermediate gray matter spinal cord injury model for future cell transplantation work with detailed behavioral readouts.

DATA COLLECTED: Abbreviated Methods: (detailed information in the Methodology document) Female Fisher 344 rats (180-200g, ~10 weeks of age) were used for these studies. Group 1 was a study with thoracic T12 laminectomy with 1-2 bilateral kainic acid (KA) spinal cord injections (n=5, saline controls n=4) followed by behavioral testing (BBB score and subscore, uneven ladder slips) at 14 days post-injury. Group 2 was a study with thoracic T13 laminectomy with 3 bilateral kainic acid spinal cord injections (n=7, saline controls n=7) followed by behavioral testing (BBB score and subscore, Even Ladder Score, Even Ladder Slips, Uneven Ladder Score, Uneven Ladder Slips, Inclined Beam Time, Inclined Beam Score, Inclined Beam Steps, Inclined Beam Completions, Hargreaves, von Frey1.4g, von Frey 60g, pLDA CatWalk Score, CatWalk AB Seq, CatWalk Body Speed, CatWalk Duty Cycle, CatWalk Forepaw Stride Length, CatWalk Forepaw Swing Time, CatWalk Hindpaw Base of Support, CatWalk Hindpaw Stride Length, CatWalk Max Contact at, CatWalk Regularity Index, CatWalk CA Sequence, CatWalk Forelimb Stand Time, CatWalk Hindlimb Stand Time, CatWalk Forelimb Duty Cycle, CatWalk Hindlimb Duty Cycle, CatWalk Hindpaw Swing Time) at 14 days post-injury. Group 3 was a study with thoracic T13 laminectomy with 3 bilateral kainic acid spinal cord injections (KA n=3, saline controls n=3) followed by behavioral testing (BBB score and subscore) at 90 days post-injury.

CONCLUSIONS: Lesions in the mixed cohort (Group 1) which we found to be primarily in spinal levels T13/L1 and not overlapping into spinal L2 did not show significant gross hindlimb and coordination deficits two weeks after injury. The BBB score of the control group was 20.63 ± 0.38 (mean ± standard error of mean) and the KA group was 18.20 ± 0.92 and not significantly different (p = 0.0642, unpaired Welch’s t-test, n = 4 control and n = 5 KA animals). The BBB subscore for the controls was 12.75 ± 0.25 and for the KA animals was 10.8 ± 0.80 and was also not significantly different (p = 0.0750; unpaired Welch’s t-test, n = 4 control and n = 5 KA animals). Two weeks post-lesioning, the percent hindlimb slips for the controls was 1.19 ± 0.79% and for the KA group was 4.29 ± 2.24% (p = 0.2470, unpaired Welch’s t-test, n = 4 controls and n = 5 KA animals). In lesions targeting spinal levels L2-L4 with 3 bilateral kainic acid injections (Group 2), gross hindlimb function after 14 days was found (BBB scores: control = 17.57 ± 0.69, KA = 11.21 ± 2.25) as well significant deficits in hindlimb function dependent on coordination (BBB subscore control = 11.29 ± 0.18, KA = 4.57 ± 1.67). KA rats show a significantly higher number of hindlimb slips on both the even and uneven horizontal ladders indicating deficits in rhythmic walking (even ladder controls = 0.70 ± 0.34%, KA = 39.91 ± 15.81%) and coordination (uneven ladder controls = 1.71 ± 0.91%, KA = 48.68 ± 15.12%). Only 3 of the 7 KA rats were able to complete the inclined beam test (controls = 100 ± 0% completion, KA = 23.81 ± 14.02%). Those KA rats that did complete the beam had a lower performance score (Controls = 95.66 ± 1.25%, KA = 21.19 ± 13.32%) further indicating deficiencies in coordination and balance. From the CatWalk gait analysis, 9 SCI parameters were combined into a pLDA score which showed that KA rats have a significantly lower pLDA score two weeks after SCI (controls = 0.13 ± 0.01, KA = 0.076 ± 0.01). Similar lesion as Group 2, Group 3 was the 90-day study that examined if KA deficits showed sustained deficits (KA n=3, saline controls n=3), BBB score controls = 19.83 ± 0.60, KA = 10 ± 2.5; BBB subscore controls = 12 ± 0.58, KA = 2.33 ± 2.33). This work shows that intermediate gray matter (laminae V-VIII) lesions spanning L2-L4 specifically lead to a stable spinal cord injury with specific behavioral readouts for further examination of cell transplantation in restoring motor function.

Female C57BL6/J mice exhibit increased heat hyperalgesia and mechanical allodynia compared to males following a T9 contusion spinal cord injury as measured by Hargreaves and SUDO von Frey testing

DOI:10.34945/F5SW2F

DATASET CITATION:

Lee S. E., Greenough E. K., Oancea P., Scheinfeld A. R., Douglas A. M., Aldrich J. C., Gaudet A. D. (2023) Female C57BL6/J mice exhibit increased heat hyperalgesia and mechanical allodynia compared to males following a T9 contusion spinal cord injury as measured by Hargreaves and SUDO von Frey testing. Open Data Commons for Spinal Cord Injury. ODC-SCI:893 http://dx.doi.org/10.34945/F5SW2F

ABSTRACT:

STUDY PURPOSE: The purpose of this study was to test if contusion spinal cord injury (SCI) causes acute-to-chronic neuropathic pain-like symptoms in mice and to determine if there are sex differences in these SCI-induced pain-like behaviors. An in-depth analysis and discussion of these data was published in DOI: 10.1089/neu.2022.0482

DATA COLLECTED: The published study is the result of a comprehensive analysis of over 100 male and female 6-20 week old C57BL6/J mice pooled from several independent studies. All mice received either a sham (T9 laminectomy) or SCI surgery (T9 laminectomy followed by a 60-75 kdyn spinal cord contusion). Locomotor recovery was assessed via the Basso mouse scale (BMS), heat hyperalgesia in hindpaws was assessed via Hargreaves test (withdrawal latencies were measured for left and right hindpaws over three trials and averaged together into a single value), and mechanical allodynia in hindpaws (left and right averaged together) was assessed via the simplified up-down (SUDO) von Frey method. In general, BMS was performed at 1, 4, 7, 10, and 14 dpo (days post operation) and every 7 days thereafter, while SUDO and Hargreaves testing was performed weekly starting at 7 dpo. Initially, a range (60, 65, and 75 kdyn) of target forces was used to determine an appropriate contusion force to use in conjunction with Hargreaves and von Frey testing. 60 and 65 kdyn had comparable results and were pooled together while 75 kdyn resulted in confounding technical issues related to hindpaw placement (see CONCLUSIONS below)—consequently, 60 kdyn was ultimately chosen for the larger sex differences study. Overall, this data set includes pre-surgery testing data and at least 3 (usually 6+) post-surgery BMS, SUDO von Frey, and/or Hargreaves data points for 170 mice (48 SCI and 44 sham females; 38 SCI and 40 sham males).

CONCLUSIONS: In the preliminary study, mechanical and heat hypersensitivity was observed in female SCI mice relative to sham mice regardless of the contusion force used, with a significant decrease in Hargreaves latency being observed in 75 vs 60-65 kdyn suggesting enhanced heat hypersensitivity. However, 75 kdyn SCI caused excess motor deficits (e.g. impaired hindpaw plantar placement) that confounded pain sensitivity measurement at acute times (i.e. 7 and 14 dpo) leaving 60 kdyn as the better option for subsequent studies. In the larger sex differences study, we note that both males and females exhibit mechanical and heat hypersensitivity (measured by SUDO von Frey and Hargreaves test respectively) post-SCI relative to sham controls. For example, at 7 dpo SCI mice (males and females combined) have an average (± SEM) Hargreaves withdrawal latency of 11.2 ±0.6 sec compared to 15.0 ±0.4 sec for shams. Similarly, at 7 dpo SCI mice have an average SUDO von Frey threshold of 6.5 ±0.2 vs. 7.9 ±0.1 for shams. Although no sex differences were observed in locomotor recovery (measured via BMS), females display amplified SCI-induced mechanical hypersensitivity relative to males at 28 dpo (average SUDO score of 7.0 ±0.2 vs. 7.8 ±0.2) as well as a consistent ~3 sec decrease in thermal withdrawal latency from 7-28 dpo compared to males (e.g. males averaged across all dpo had a Hargreaves score of 12.2 ±0.5 sec vs. 9.2 ±0.3 sec for females).

Developmental stage of transplanted neural progenitor cells influences anatomical and functional outcomes after C5 dorsal column lesion and T12 contusion spinal cord injury in male and female C57BL/6 mice

DOI:10.34945/F5TS33

DATASET CITATION:

Dulin J. N., Aceves M., McCreedy D. A. (2023) Developmental stage of transplanted neural progenitor cells influences anatomical and functional outcomes after C5 dorsal column lesion and T12 contusion spinal cord injury in male and female C57BL/6 mice. Open Data Commons for Spinal Cord Injury. ODC-SCI:873 http://dx.doi.org/10.34945/F5TS33

ABSTRACT:

STUDY PURPOSE: Neural progenitor cell (NPC) transplantation is a promising therapeutic strategy for replacing lost neurons following spinal cord injury (SCI). However, how graft cellular composition influences regeneration and synaptogenesis of host axon populations, or recovery of motor and sensory functions after SCI, is poorly understood. We transplanted developmentally-restricted spinal cord NPCs, isolated from E11.5-E13.5 mouse embryos, into sites of adult mouse SCI and analyzed graft axon outgrowth, cellular composition, host axon regeneration, and locomotor and thermal sensory behavior.

DATA COLLECTED: Abbreviated Methods: (More detailed information can be found in the Methodology document.) Male and female C57BL/6 mice (18-33 g) were used for this study. Either cervical (C5) dorsal column lesion SCI or thoracic (T12) contusion SCI (50 kdynes, Infinite Horizon impactor device) was performed. Either immediately after SCI (C5 dorsal column lesion groups) or 2 weeks after SCI (T12 contusion groups), mouse spinal cord neural progenitor cells of specific developmental stages or fibrin/thrombin matrix was injected into the lesion site. The specific developmental stages include: E11.5 NPCs, E12.5 NPCs, E13.5 NPCs, E12.5 dorsal NPCs, and E13.5 ventral NPCs. For the animals receiving C5 dorsal column lesion, they were sacrificed at 4 weeks post-transplantation for histological analysis. No behavioral testing was performed on these animals. For the animals receiving T12 contusion, we conducted behavioral assessments both prior to SCI and until 70 days post-SCI, then performed histological assessments of graft survival. Behavioral assessments include open field locomotor (BMS) testing and Hargreaves (thermal sensitivity) testing of hindpaws. BMS was performed at 1, 3, 5, 7 days post-SCI and weekly until 70 DPI. Hargreaves testing was performed weekly at 14-70 DPI.

CONCLUSIONS: -Immunohistochemical analysis showed that the abundance of some subtypes of neurons are significantly different between graft types. -Axon outgrowth is significantly greater in E11.5 grafts than all other groups. -5-HT+ axon regeneration into E11.5 grafts is significantly greater than other graft types. -CGRP+ axon regeneration into E13.5 grafts is significantly greater than other graft types. -There was no effect of any graft type on hindlimb locomotor recovery compared to vehicle (fibrin/thrombin matrix). -Animals with E13.5 grafts exhibited significantly lower thermal latency scores compared to all other treatment groups. Together, these findings indicate that NPC graft cellular composition varies with the developmental stage of donor cells, and that anatomical and behavioral outcomes are influenced by graft type.

hM4Di-mediated chemogenetic attenuation of acute nociceptive signaling enhances locomotor function and prevents the development of thermal hypersensitivity following moderate T10 spinal cord contusion injury in female Sprague-Dawley rats

DOI:10.34945/F5730S

DATASET CITATION:

Amar Kumar P., Dulin J. N. (2023) hM4Di-mediated chemogenetic attenuation of acute nociceptive signaling enhances locomotor function and prevents the development of thermal hypersensitivity following moderate T10 spinal cord contusion injury in female Sprague-Dawley rats. Open Data Commons for Spinal Cord Injury. ODC-SCI:851 http://dx.doi.org/10.34945/F5730S

ABSTRACT:

STUDY PURPOSE: Identifying novel therapeutic approaches to promote recovery of neurological functions following spinal cord injury (SCI) remains a great unmet need. Nociceptive signaling in the acute phase of SCI has been shown to inhibit recovery of locomotor function and promote the development of chronic neuropathic pain. We therefore hypothesized that inhibition of nociceptive signaling in the acute phase of SCI might improve long-term functional outcomes in the chronic phase of injury. To test this hypothesis, we took advantage of a selective strategy utilizing AAV6 to deliver inhibitory (hM4Di) DREADDs to nociceptors of the L4-L6 dorsal root ganglia, in order to evaluate the effects of transient nociceptor silencing on long-term sensory and motor functional outcomes in a rat thoracic contusion SCI model.

DATA COLLECTED: Abbreviated Methods: (More detailed information can be found in the Methodology document.) Female Sprague-Dawley rats (200-250g) were used for this study. AAV6-hM4Di or AAV6-YFP (8x10^12 genome copies/mL) was injected into bilateral sciatic nerves 4 weeks prior to SCI/sham. Either thoracic (T10) contusion SCI (150 kydnes, 1 s dwell, Infinite Horizon Impactor device) or sham surgery was performed at Day 0. Immediately following SCI, either clozapine-N-oxide (CNO; hM4Di agonist) or vehicle (0.05% DMSO) was injected intraperitoneally. Beginning on the day of surgery, either CNO or vehicle + 5 mM saccharine was administered in drinking water for 14 days. Following CNO or vehicle treatment from 0-14 days post-SCI, we conducted behavioral assessments until 70 days post-SCI, then performed histological assessments of lesion severity and axon plasticity. Behavioral assessments include open field locomotor (BBB) testing, CatWalk gait analysis, Hargreaves (thermal sensitivity) testing of hindpaws, von Frey (mechanical reactivity) testing of hindpaws, and acetone (cold allodynia) testing of hindpaws. All behavioral testing except for BBB was performed weekly beginning at 2 weeks post-SCI and continuing until 10 weeks post-SCI. BBB data were collected at 1, 3, 7 days post-SCI then weekly until 10 weeks post-SCI. At the study endpoint, animals were transcardially perfused and spinal cord and DRG tissue was collected for histological analysis. The following histological measures were assessed: Lesion volume based on GFAP immunoreactivity, grey matter sparing based on NeuN labeling, white matter sparing based on beta-tubulin labeling, intensity of beta-tubulin immunoreactivity, density of CGRP+ fibers in the T10 and L4 spinal cord, density of IB4-binding fibers in the T10 and L4 spinal cord, density of synaptophysin+ punctae on ChAT+ motor neurons in the L4 spinal cord. Detailed histological analysis methods are described in the Methodology file. Results: -Immunohistochemistry results show highly selective expression of hM4Di within small diameter nociceptors including CGRP+ and IB4-binding neurons. -A two-way repeated measures ANOVA identified a significant effect of time x treatment (P<0.0001) in the thermal hindlimb paw withdrawal scores between SCI-YFP and SCI-hM4Di animals. At the first time point tested following SCI (14 DPI), animals in the SCI-YFP group showed significantly reduced withdrawal latencies to a thermal stimulus relative to naïve controls, and this continued until 35 DPI (Tukey's multiple comparisons test, P<0.05 at each time point). -No significant differences in von Frey mechanical reactivity scores, or acetone cold allodynia scores, were observed between SCI-YFP and SCI-hM4Di treatment groups. -A repeated measures ANOVA identified a significant effect of treatment on open field locomotor (BBB) scores (P=0.0489), indicating that acute nociceptor silencing improved long-term locomotor recovery. -Based on a pLDA analysis of CatWalk data, we found that compared to the naïve and sham groups, both SCI groups exhibited significantly lower pLDA scores at all timepoints post-SCI (P < 0.0002 in all cases); however, the pLDA scores of SCI groups were not significantly different to each other. -Lesion volume, calculated by drawing regions of interest around GFAP+ reactive glial cell layers immediately surrounding the lesion cavity, was not significantly different between SCI-YFP and SCI-hM4Di subjects. -We found that density of both CGRP+ and IB4-binding axons did not differ significantly between SCI groups.

CONCLUSIONS: Together, these findings suggest that nociceptor silencing early after SCI may promote beneficial plasticity in the acute phase of injury that can impact long-term functional outcomes, and support previous work highlighting primary nociceptors as possible therapeutic targets for pain management after SCI.

CatWalk Gait parameters at 8 weeks after T10 thoracic contusion using the MASCIS impactor model III in male and female 8- to 10-week old rats

DOI:10.34945/F5VS3D

DATASET CITATION:

Liu S., Xing C., Wei H., Guo J., Wang L., Li B., Ma H., Zhong H., Zhou M., Zhu S., Zhu R., Ning G. (2022) CatWalk Gait parameters at 8 weeks after T10 thoracic contusion using the MASCIS impactor model III in male and female 8- to 10-week old rats. Open Data Commons for Spinal Cord Injury. ODC-SCI:822 http://dx.doi.org/10.34945/F5VS3D

ABSTRACT:

STUDY PURPOSE: In preclinical studies of spinal cord injury (SCI), behavioral assessment is crucial in determining the effectiveness of interventions. As an automated gait analysis system, catwalk provides a large number of objective gait parameters through the acquisition and calculation of rodent footprints and videos. However, the gait parameters may not all be suitable for assessing hindlimb locomotor function in animals with thoracic contusion SCI. The purpose of this study was to discover CatWalk gait parameters applicable to different severity ranges (sham, mild, moderate, severe) of T10 spinal cord contusion injury in rats.

DATA COLLECTED: A total of 48 adult male and female Wistar rats, weighing 190-210 g and aged 8-10 weeks old, were used in this study. The thoracic spinal cord of rats was contused by MASCIS impactor model III (W. M. Keck Center, Rutgers University, United States) according to the modified Allen's method. The animals subsequently underwent T10 spinal cord contusions caused by a 10 g impactor dropped at varying heights of 12.5 mm (mild), 25.0 mm (moderate), and 50.0 mm (severe). Animals in the sham group only received laminectomy without contusion.N=12/group. Catwalk gait analysis were performed at 8 weeks post-injury. The performance of moving in one direction without stopping or turning around was recorded.The specific detection settings used in the experiment were as follows: camera gain = 17.75 dB, green intensity threshold = 0.12, red ceiling light= 17.1 V, and green walkway light = 15.4 V. Multiple groups were compared using Kruskal-Wallis test with Dunn's post-hoc test.

CONCLUSIONS: Among the CatWalk gait parameters, regularity index could reflect differences between the Sham, SCI-mil, and SCI-mod groups, but not between the SCI-mod and SCI-sev groups . Our work provides a comprehensively CatWalk gait parameters for SCI studies, which could be applicable to objectively reflect locomotor recovery of T10 spinal cord contusion injury in rats.

Facilitation of sensory axon conduction to motoneurons during cortical or sensory evoked primary afferent depolarization (PAD) in humans

DOI:10.34945/F5WS3Q

DATASET CITATION:

Metz K., Concha-Matos I., Li Y., Afsharipour B., Thompson C. K., Negro F., Bennett D., Gorassini M. A. (2022) Facilitation of sensory axon conduction to motoneurons during cortical or sensory evoked primary afferent depolarization (PAD) in humans. Open Data Commons for Spinal Cord Injury. ODC-SCI:811 http://dx.doi.org/10.34945/F5WS3Q

ABSTRACT:

STUDY PURPOSE: Sensory feedback to the spinal cord is essential for motor control. After injury to the central nervous system, such as spinal cord injury, the mechanisms that regulate sensory feedback become unbalanced leading to poor motor control, spasticity and pain. Unfortunately, our understanding of sensory feedback is relatively limited, even in the uninjured nervous system, making the treatment of motor control issues related to sensory feedback challenging. It was previously believed that sensory feedback was regulated by specialized GABAergic neurons that inhibited sensory inputs in the spinal cord, termed presynaptic inhibition. Recent findings in rodents shows that these GABAergic pathways actually facilitate conduction along afferents in the spinal cord by depolarizing the afferent at vulnerable branch points and reducing branch point failure. Based on these recent findings, we evaluated similar pathways in uninjured humans, with future hopes of evaluating these pathways in participants with spinal cord injury.

DATA COLLECTED: The Ia - motoneuron pathway was measured using an H-reflex, evoked below half the maximum amplitude in either the soleus muscle (stimulation of the tibial nerve at the popliteal fossa) or the abductor hallucis (AbHal) muscle (stimulation of the tibial nerve at the popliteal fossa or at the medial malleolus). The peak-peak amplitude of the H-reflex was measured before (test) and after (cond) a conditioning stimulation to putative active GABAergic networks in the spinal cord and primary afferent depolarization (PAD) in the test (soleus or AbHal) afferents. Presynaptic effects of the conditioning stimulation on the test Ia afferents were discerned from postsynaptic effects on the test motoneurons using single motor unit and EMG analysis with the conditioning stimulation applied alone, thus giving information about the isolated postsynaptic effects of the conditioning stimulation. The probability of discharge of a tonically firing soleus single motor unit at the latency of the H-reflex (after tibial nerve stimulation) was also compared before and after a conditioning stimulation to putative evoked PAD. Specifically, the cutaneous branch of the superficial fibular nerve (SFN) was electrically stimulated on the top of the foot at an intensity just above perception threshold at various interstimulus intervals (ISIs) between 0 and 300 ms to assess short-lasting phasic PAD (200 Hz, 10 ms) and at longer intervals (out to 130 seconds) to assess long-lasting tonic PAD (0.2 and 2 Hz for 10s, 200 Hz for 500 ms) using the soleus H-reflex. Similarly, the common fibular nerve (CFN) was electrically stimulated (near the fibular head on the ipsilateral leg) at motor threshold (1.0 x MT) 0-300 ms before activation of the soleus H-reflex. A heteronymous H-reflex in the Biceps Hetero (stimulation of the tibial nerve supplying the medial gastrocnemius) was also conditioned by SFN and CFN stimulation. Activation of the corticospinal tract (CST) using transcranial magnetic stimulation (TMS; 0.9 x active motor threshold) of the contralateral motor cortex was used to condition the soleus and AbHal H-reflex and assess short-lasting phasic PAD. A peristimulus frequencygram (PSF) of the firing rate of a tonically discharging single motor unit in response to the putative PAD-evoking conditioning stimulation over many trials was created, with the time of the conditioning stimulation time-locked to 0 ms. The PSF was binned into 20 ms averages and the average firing rate at each bin was compared to the time when an H-reflex would have occurred at the various ISIs. A similar analysis was done for the EMG recorded during a small contraction. Self-facilitation of the soleus H-reflex was also measured by comparing H-reflexes evoked every 5 seconds. The average peak-peak amplitude of the first 7 H-reflexes were compared to the peak-peak amplitude of the next 7 H-reflexes. A more detailed description of the data collected can be found in the accompanying paper.

CONCLUSIONS: We find new evidence that sensory and corticospinal pathways known to activate GABAergic networks in the spinal cord can facilitate H-reflexes without any accompanying faciliatory postsynaptic effects.

Immune phenotyping SCI patients versus healthy controls

DOI:10.34945/F5588X

DATASET CITATION:

Fraussen J., Beckers L., van Laake-Geelen C. C.M.., Depreitere B., Deckers J., Cornips E. M.J.., Peuskens D., Somers V. (2022) Immune phenotyping SCI patients versus healthy controls. Open Data Commons for Spinal Cord Injury. ODC-SCI:774 http://dx.doi.org/10.34945/F5588X

ABSTRACT:

STUDY PURPOSE: Following a spinal cord injury (SCI), an inflammatory immune reaction is triggered which results in advanced secondary tissue damage. The systemic post-SCI immune response is poorly understood. This study aimed to extensively analyse the circulating immune cell composition in traumatic SCI patients in relation to clinical parameters.

DATA COLLECTED: High-dimensional flow cytometry was performed on peripheral blood mononuclear cells of 18 traumatic SCI patients and 18 healthy controls to determine immune cell subsets. SCI blood samples were collected at multiple time points in the (sub)acute (0 days to 3 weeks post-SCI, (s)aSCI) and chronic (6 to >18 weeks post-SCI, cSCI) disease phase. Not all time points were available for every included SCI patient due to organizational issues (late inclusion) or withdrawal of subjects from the study. Twelve SCI patients presented with a cervical injury, 5 with a thoracic injury and 1 with a lumbar injury. We first analyzed the major immune cell subsets in PBMC of HC, (s)aSCI patients and cSCI patients. These included CD14+ monocytes, natural killer (NK) cells, which are classically divided into cytotoxic CD56loCD16+ and highly cytokine producing CD56hiCD16+/- subsets, CD19+ B cells and CD3+ T cells, divided into CD4+ helper and CD8+ cytotoxic subsets. Next, the following subsets were studied in both CD4+ and CD8+ T cells: naive (Tnaive, CD45RA+CD45RO-), memory (Tm, CD45RA-CD45RO+), effector memory (Tem, CD45RA-CCR7-), central memory (Tcm, CD45RA-CCR7+), effector memory re-expressing CD45RA (Temra, CD45RA+CCR7-) and regulatory (Treg, CD25+CD127low). CD19+ B cells were studied in more detail by analysis of CD24hiCD38hi transitional, IgD+CD27- naive, IgD+CD27+ non class-switched memory (NCSM), IgD-CD27+ class-switched memory (CSM), IgM+IgD-CD27+ IgM only and IgD-CD27- double negative (DN) subsets. IgM, IgG and IgA were included to further define memory responses.

CONCLUSIONS: T cell frequencies were increased in cSCI patients. Both CD4+ T cells and B cells were shifted towards memory phenotypes in (s)aSCI patients and cSCI patients, respectively. Most profound changes were observed in the B cell compartment. Decreased immunoglobulin (Ig)G+ and increased IgM+ B cell frequencies reflected disease severity, as these correlated with American Spinal Injury Association (ASIA) impairment scale (AIS) scores. Post-SCI B cell responses consisted of an increased frequency of CD74+ cells and CD74 expression level within total B cells and B cell subsets. Findings from this study suggest that post-SCI inflammation is driven by memory immune cell subsets. The increased CD74 expression on post-SCI B cells could suggest the involvement of CD74-related pathways in neuroinflammation following SCI. In addition, the clinical and prognostic value of monitoring circulating IgM+ and IgG+ B cell levels in SCI patients should be further evaluated.

Contralateral axon sprouting in male and female c57Bl/6J mouse cervical spinal cord by uninjured corticospinal axons following unilateral pyramidotomy and cortical injection of kinase inhibitors

DOI:10.34945/F5T01D

This dataset has been removed from public view at the request of the authors, as the underlying records have been found to be insufficient. The authors regret any inconvenience caused.

Lung immunity after complete thoracic spinal cord transection in female mice

DOI:10.34945/F52G69

DATASET CITATION:

Mifflin K. A., Brennan F. H., Guan Z., Kigerl K. A., Filous A. R., Mo X., Schwab J. M., Popovich P. G. (2022) Lung immunity after complete thoracic spinal cord transection in female mice. Open Data Commons for Spinal Cord Injury. ODC-SCI:735 http://dx.doi.org/10.34945/F52G69

ABSTRACT:

STUDY PURPOSE: Pulmonary infection is a leading cause of morbidity and mortality after spinal cord injury (SCI). Although SCI causes atrophy and dysfunction in primary and secondary lymphoid tissues with a corresponding decrease in the number and function of circulating leukocytes, it is unknown if this SCI-dependent systemic immune suppression also affects the unique tissue-specific antimicrobial defense mechanisms that protect the lung. Here, we test the hypothesis that SCI directly impairs pulmonary immunity and subsequently increases the risk for developing pneumonia.

DATA COLLECTED: The dataset includes n=242 C57/Bl6 female mice aged 9-13 weeks. Mice received a complete transection SCI at either the thoracic level 1 (T1Tx) or 3 (T3Tx). Sham mice underwent a laminectomy only at the appropriate thoracic level. Flow cytometry was used to assess lung immune cells (neutrophils, T- and B-lymphocytes, alveolar macrophages, CD45+ immune cells) at 12h ((n=8-12 T3Tx, n=8-12 T3 sham), 3d (n=12 T3Tx, n=12 T3 sham, n=7 T1Tx, n=7 T1 sham), and 28d (n=12 T3Tx, n=12 T3 sham) post- injury. Corresponding cultures of lung homogenate on blood agar (n=7 12h & 3dpi T3Tx and 3dpi T3 Sham, n=12-14 28dpi T3 Sham mice, n=12-14 28dpi T3Tx, n=18 T1Tx, n=13 T1 Sham) and CHROMagar ™ plates (n=18 T1Tx, n=13 T1 Sham) at these timepoints were also used to assess pulmonary bacterial burden after injury. The drug AMD3100, a CXCR4 antagonist (1mg/mouse given subcutaneously 1h post-injury then daily for either 3d or 7d), was also tested in T1Tx mice to see if the drug improved pulmonary immune function as measured by flow cytometry (n=10 T1Tx vehicle and n= 13 T1Tx AMD3100 mice), altered blood immune cell counts (n=8 T1 sham, n=9 T1Tx vehicle and n=10 T1Tx AMD3100) or reduced bacteria load (n=21 T1Tx vehicle and n=23 T1Tx AMD3100 for 3dpi studies, n=9 T1Tx vehicle and n=10 T1Tx AMD3100 for 7dpi studies). A subset of AMD3100 treated mice also underwent an induced pneumonia procedure (n=11 T3Tx vehicle + pneumonia and n=11 T3TxAMD3100 + pneumonia). Finally, n=4 mice were pooled from T3 Sham or T3Tx groups to perform a PCR array for lung immunity genes and RT-qPCR on 10 identified hub genes at 12h and 3d post-injury.

CONCLUSIONS: Using mouse models of severe high thoracic level 1 or 3 SCI, we find that recruitment of circulating leukocytes and transcriptional control of immune signaling in the lung is impaired after SCI, creating an environment that is permissive for infection. Specifically, we see a sustained loss of pulmonary leukocytes (up to ~60% loss), a loss of alveolar macrophages (30% loss) at chronic timepoints post-injury, and a decrease in immune modulatory genes, especially cytokines (e.g. Tnf, Il6, Infg), needed to eliminate pulmonary infections. Importantly, this injury-dependent impairment of pulmonary antimicrobial defense is only partially overcome by boosting the recruitment of immune cells to the lung with the drug AMD3100, an FDA-approved drug that mobilizes leukocytes and hematopoietic stem cells from bone marrow. Collectively, these data indicate that the immune suppressive effects of SCI extend to the lung, a unique site of mucosal immunity. Furthermore, preventing lung infection after SCI will likely require novel strategies, beyond the use of orthodox antibiotics, to reverse or block tissue-specific cellular and molecular determinants of pulmonary immune surveillance.

Genetic deletion of the glucocorticoid receptor in Cx3cr1+ myeloid cells is neuroprotective and improves motor recovery in female mice after 75 kdyne T9 contusion spinal cord injury

DOI:10.34945/F56887

DATASET CITATION:

Madalena K. M., Brennan F. H., Popovich P. G. (2022) Genetic deletion of the glucocorticoid receptor in Cx3cr1+ myeloid cells is neuroprotective and improves motor recovery in female mice after 75 kdyne T9 contusion spinal cord injury. Open Data Commons for Spinal Cord Injury. ODC-SCI:734 http://dx.doi.org/10.34945/F56887

ABSTRACT:

STUDY PURPOSE: Glucocorticoid receptors (GRs), part of the nuclear receptor superfamily of transcription factors (TFs), are ubiquitously expressed in all cell types and regulate cellular responses to glucocorticoids (e.g., cortisol in humans; corticosterone in rodents). In myeloid cells, glucocorticoids binding to GRs can enhance or repress gene transcription, thereby imparting distinct and context-dependent functions in macrophages at sites of inflammation. In experimental models and in humans, glucocorticoids are widely used as anti-inflammatory treatments to promote recovery of function after SCI. Thus, we predicted that deleting GR in mouse myeloid lineage cells (i.e., microglia and monocyte-derived macrophages) would enhance inflammation at the site of injury and worsen functional recovery after traumatic spinal cord injury (SCI).

DATA COLLECTED: The dataset includes n=29 female mice with genotype of GRf/f (Macrophage GR intact) or Cx3cr1-Cre;GRf/f (Macrophage GR deleted). Mice received a 75 kdyne T9 IH contusion SCI or sham (laminectomy only) surgery. The Basso Mouse Scale (BMS) main score and sub-score were used to assess motor function. At 35 dpi, animals were perfused with 0.1M PBS followed by 4% PFA, and spinal tissues were cryopreserved in OCT at -80oC. Tissues were cut in serial sections on a cryostat, 12 um thick along the coronal axis. Spared myelin (EC), spared axons (NFH), and vessel sparing (CD31) were assessed using immunohistochemical staining. GFAP, Iba1, MBP, CD68, and Oil Red O staining was performed in coronal spinal histological sections at 35 days post-injury (dpi). Injury parameters are included for all injured mice. For validation of GR knockout in myeloid cells, we used additional male and female uninjured Cx3cr1-Cre;TdT mice (n=6) and uninjured TdTomato controls (n=4).

CONCLUSIONS: Contrary to our prediction, the intraspinal macrophage response to a moderate (75 kdyne) spinal contusion SCI was reduced in Cx3cr1-Cre;GRf/f conditional knockout mice (with GR specifically deleted in myeloid cells). This phenotype was associated with improvements in hindlimb motor recovery (increased BMS motor scores and subscores; 57% some-most coordination in macrophage GR deleted vs. 0% in macrophage GR intact, 71% paws parallel in macrophage GR deleted vs. 13% in macrophage GRintact, and 43% severe trunk in macrophage GR deleted vs. 62% in macrophage GR intact). GR depletion in myeloid cells also improved myelin sparing (47% more erichrome cyanine staining in lesion epicenter), axon preservation/regeneration (34% more neurofilament staining in lesion epicenter), and microvascular protection/plasticity (33% more CD31 staining) relative to SCI mice with normal myeloid cell GR expression. Further analysis of phagocytic macrophage morphology and myelin basic protein engulfment using CD68 and MBP staining revealed that macrophage GR deletion impairs lipid and myelin phagocytosis and presence of macrophages with a 'foamy' morphology. Together, these data reveal endogenous GR signaling as a key pathway that normally inhibits mechanisms of macrophage-mediated repair after SCI.

Neuroprotection and bromodomain and extra-terminal domain (BET) protein inhibition after T8 spinal cord contusion in female rats and mice

DOI:10.34945/F5ZP4P

DATASET CITATION:

Cerqueira S. R., Benavides S., Lee H. E., Ayad N. G., Lee J. K. (2022) Neuroprotection and bromodomain and extra-terminal domain (BET) protein inhibition after T8 spinal cord contusion in female rats and mice. Open Data Commons for Spinal Cord Injury. ODC-SCI:696 http://dx.doi.org/10.34945/F5ZP4P

ABSTRACT:

STUDY PURPOSE: In this study, we tested the therapeutic potential of inhibiting bromodomain and extraterminal domain (BET) proteins after SCI, and investigated the role of the BET protein BRD4 in macrophages during progression of SCI pathology.

DATA COLLECTED: We investigated the anti-inflammatory and neuroprotective effects of BET inhibition in a rat moderate T8 contusion model (IH impactor; 200 kDyn). First, we performed a dose response study (7.5; 25; 75 mg/kg) of two compounds (IBET-151; IBET-762) administered intraperitoneally (IP) at 3 hours post-SCI. At 6 hours post-SCI, 4-mm long samples of spinal cord centered at the injury epicenter were collected to measure expression of proinflammatory cytokines using qPCR (n=6/group). We then performed IP administration of vehicle, IBET-151 or IBET-762 (50mg/kg) at 3 and 18 hours after SCI and verified transcription levels of proinflammatory cytokines at 24 hours post-SCI using RT-qPCR (n=6/group). Based on these data, we performed a long-term study using the same experimental SCI model, and treated animals with vehicle or IBET-762 (3h; 18h; 1, 2 and 3 days after SCI) (n=11-12/group). Rats were scored using the Basso, Beattie and Breshnahan (BBB) scale at day 1, and weekly thereafter. Eight weeks after SCI, animals were perfused, spinal cord injury sites removed and sectioned sagittally. Unbiased stereological quantifications of lesion size (GFAP negative area), fibrotic scar formation (PDGFRb positive area), and immune cell infiltration (CD11b positive area) in the lesion site were done in 3-5 sections per animal. Next, we investigated the role of BRD4 in macrophages after SCI using LysM-Cre-TDT BRD4 KO mice. Female mice (8-10 weeks old) were subjected to a T8 contusion (IH impactor; 65 kDyn) (WT controls n=8; Brd4 KO n=8). Locomotor recovery was assessed at day 1 after SCI and weekly thereafter using the Basso Mouse Scale open field test for mice. Four weeks after SCI, mice were perfused and spinal cord injury sites collected and sectioned sagittally for histological evaluation. Unbiased stereological quantifications of lesion size (GFAP negative area), and macrophage infiltration (TDT positive area) in the lesion site were done in 3-5 sections per animal. Finally, we evaluated the effect of macrophage-targeted BET inhibition using compound delivery via liposomes. Female mice (8-10 weeks old) were subjected to a T8 contusion (IH impactor; 65 kDyn) and liposomes were administered IP at days 1, 3, 6, 11, and 18 (vehicle liposomes n=5; IBET-762 liposomes n=5). Locomotor recovery was assessed at day 1 after SCI and weekly thereafter using the Basso Mouse Scale open field test for mice. Four weeks after SCI, mice were perfused and spinal cord injury sites collected and sectioned sagittally for histological evaluation. Unbiased stereological quantifications of lesion size (GFAP negative area), fibrotic scar (PDGFRb positive area), and macrophage infiltration (CD11b positive area) in the lesion site were done in 3-5 sections per animal.

CONCLUSIONS: Taken together, our data indicate that systemic I-BET762 treatment is neuroprotective, and the histopathological improvement observed is likely to be a result of effects on non-macrophage targets.

Studying Sex as a Biological Variable: Considerations of weight, anemia and mortality following thoracic spinal cord contusion injury in male and female mice

DOI:10.34945/F5D01P

DATASET CITATION:

Stewart A. N., MacLean S. M., Stromberg A. J., Whelan J. P., Bailey W. M., Wilson M. E., Gensel J. C. (2021) Studying Sex as a Biological Variable: Considerations of weight, anemia and mortality following thoracic spinal cord contusion injury in male and female mice. Open Data Commons for Spinal Cord Injury. ODC-SCI:652 http://dx.doi.org/10.34945/F5D01P

ABSTRACT:

STUDY PURPOSE: The purpose of this study was to examine functional recovery, morbidity, mortality, etc. as a function of both sex and age in C57Bl/6J mice after thoracic contusion SCI (T9, 60kdyn, IH). Experimental groups consist of young and middle-aged mice (4 or 14 months old at the time of injury) of both sexes, male or female, that underwent contusion or sham injury. The data in this collection and a thorough interpretation of the data was published in manuscript form in Stewart et al., 2020 PMIDs: 32849242; 33193066: Considerations for Studying Sex as a Biological Variable in Spinal Cord Injury. Stewart AN, MacLean SM, Stromberg AJ, Whelan JP, Bailey WM, Gensel JC, Wilson ME. Front Neurol. 2020 Aug 5;11:802. doi:10.3389/fneur.2020.00802. eCollection 2020.

DATA COLLECTED: Collectively, data from 8 experimental groups: sex (2) x age (2) x injury (2). Sample sizes range from 5-10 depending upon the outcome measure and time of sacrifice. Data for this study include weights after injury, overall mortality, injury-induced anemia as determined by red blood cell ratios in plasma (28 days post injury), estrous cycle information for females (weekly for 4 weeks) along with endpoint (28 days) plasma estradiol levels, as well as, BMS scores at 1 and 28 days after injury. The data presented is a comprehensive collector of data collected over several years across different experiments utilizing both male and female mice. For this reason, not all outcomes were collected for all subjects (missing or NA is used to indicate data not collected). Not all studies used the same outcome measures (i.e. BMS) so not all outcomes were collected for each subject.

CONCLUSIONS: The data presented in this dataset show that sex contributes to a variety of factors after SCI including weight loss, anemia, and mortality. Further, age potentiates sex-dependent effects. Sex-specific accommodations, i.e. single housing, likely also impact SCI recovery. A corrigendum for the original manuscript was published to correct for some miscoded data, this data set reflects the corrected data as published in the corrigendum. Through our analyses, we identify several potential confounds that should be considered in a study design incorporating both sexes including differences in anatomy, behavior, housing, and drug metabolism. Ultimately, our ability to consider sex as a biological variable in the study of SCI will depend upon open and rigorous data reporting and interpretation.

ASIA Impairment Scale, level of injury, intraoperative time series mean arterial pressure and heart rate after spinal cord injury in patients in a multi-site retrospective TRACK-SCI cohort: site 1 of 2

DOI:10.34945/F5R59J

DATASET CITATION:

Torres-Espin A., Haefeli J., Ehsanian R., Torres D., Almeida C., Huie J., Chou A., Dirlikov B., Suen C. G., Nielson J. L., Kyritsis N., Duong-Fernandez X., Thomas L. H., Hemmerle D. D., Morozov D., Sanderson N., Talbott J., Manley G. T., Dhall S. S., Whetstone W. D., Bresnahan J. C., Beattie M. S., McKenna S. L., Pan J. Z., Ferguson A. R. (2021) ASIA Impairment Scale, level of injury, intraoperative time series mean arterial pressure and heart rate after spinal cord injury in patients in a multi-site retrospective TRACK-SCI cohort: site 1 of 2. Open Data Commons for Spinal Cord Injury. ODC-SCI:245 http://dx.doi.org/10.34945/F5R59J

ABSTRACT:

STUDY PURPOSE: The main goal for the collection of this data was to assess whether intraoperative hemodynamics (arterial pressure and heart rate) is associated to patient neurological improvement at hospital discharge in spinal cord injury (SCI). The data were collected in two level I trauma centers in Northern California (Santa Clara Valley Medical Center and Zuckerberg San Francisco General Hospital). This dataset represents the data from one of the centers, while the data collected in the other center is available elsewhere (see provenance). Both datasets have been harmonized, meaning variables and data dictionary align.

DATA COLLECTED: The dataset includes mean arterial pressure (MAP) and heart rate (HR) at 5 minute intervals during SCI associated surgery for n = 87 patients, as well as summaries across surgery time (average, standard deviation, kurtosis and skewness). The data also include demographics (age and gender), length metrics (surgery duration and days from injury to discharge), and injury characteristics and outcome metrics (AIS grade at admission, AIS grade at hospital discharge, dichotomized neurological level of injury: cervical vs. non-cervical).

CONCLUSIONS: Through joint analysis of both centers’ data and using machine learning methods we discovered a non-linear association of MAP during surgery and neurological improvement from hospital admission to hospital discharge. Moreover, we derived a MAP range [76-104/117mmHg] at which the longer the patient is outside that range (hypotension or hypertension) during surgery, the less likely is that the patient would have a robust neurological improvement (see the associated manuscript for further details).

Hemodynamic stability, incidence of pain, skin redness, autonomic dysreflexia and other safety related outcome measures of transcutaneous electrical spinal cord stimulation in children with cervical and thoracic spinal cord injuries

DOI:10.34945/F5HP4N

DATASET CITATION:

Keller A. V., Singh G., Sommerfeld J., King M., Ugiliweneza B., D’Amico J., Gerasimenko Y., Behrman A. L. (2021) Hemodynamic stability, incidence of pain, skin redness, autonomic dysreflexia and other safety related outcome measures of transcutaneous electrical spinal cord stimulation in children with cervical and thoracic spinal cord injuries. Open Data Commons for Spinal Cord Injury. ODC-SCI:465 http://dx.doi.org/10.34945/F5HP4N

ABSTRACT:

STUDY PURPOSE: This was a pilot study to establish the safety and feasibility of transcutaneous electrical spinal cord stimulation (scTS) to augment upright sitting posture and trunk control in children with spinal cord injury.

DATA COLLECTED: The primary safety-related outcome measures were assessed over the 3 days for each participant and included: hemodynamic stability assessed using brachial arm blood pressure (mmHg) and heart rate (beats per minute) at baseline (no stimulation), during stimulation, at the end of experiments and if a child presents with symptoms of autonomic dysreflexia (e.g. facial flushing, reporting of headache, etc); incidence of autonomic dysreflexia (increase in blood pressure over 20 mmHg from a previous blood pressure measurement); incidence of pain in response to stimulation (assessed using FACES Pain Scale-Revised for children under 8 and Visual Analogue Scale for children 8 and older); incidence of skin redness under the stimulating electrodes. Trunk kinematics and center of pressure displacement data are published as a separate dataset on ODC-SCI titled: "Trunk kinematic and center of pressure displacement time series data during transcutaneous lumbosacral spinal cord stimulation and passive pelvic tilt in human pediatric participants with trunk control impairments due to cervical or thoracic spinal cord injury (pilot clinical trial NCT03975634)"

CONCLUSIONS:

Treatment with Gallium Maltolate after spinal cord thoracic contusion in female mice, functional outcome and inflammatory markers

DOI:10.34945/F5S59V

DATASET CITATION:

Stehlik K. E., Pelisch N., Kurpad S. N., Swartz K. R., Kroner A. (2021) Treatment with Gallium Maltolate after spinal cord thoracic contusion in female mice, functional outcome and inflammatory markers. Open Data Commons for Spinal Cord Injury. ODC-SCI:619 http://dx.doi.org/10.34945/F5S59V

ABSTRACT:

STUDY PURPOSE: Hemorrhage after SCI is associated with worse functional outcome, compared to non-hemorrhage SCI cases. Toxicity and pro-inflammatory effects of hemoglobin, its breakdown factors, and iron deposition may contribute to this problem. Gallium nitrate and Gallium maltolate are used in cancer therapy and in treatment of bacterial infections as iron competitors, resulting in iron starvation and cell death of bacteria or cancer cells, which have a high iron demand. While this mechanism is not necessarily relevant to control tissue hemorrhage after SCI, the additional anti-inflammatory role of gallium maltolate made it an interesting treatment consideration.

DATA COLLECTED: This experiment was performed in two cohorts of female, 8-week-old C57BL/6 mice (Charles River Laboratories). Under isoflurane anesthesia, mice were subjected to a 40 kdyne contusion (Infinite Horizon Impactor) injury at T11. In the first cohort, gallium maltolate versus a saline control (n=8-10) was injected subcutaneously immediately after injury, as well as at two and at four days later (200mg/KG). In contrast to earlier reports, this dose was not well tolerated: some mice died and the experiment was terminated at day 7. In the second cohort, mice received 25mg/KG gallium maltolate or saline s.c. on day 0 (immediately after injury), and every second day for the survival period (three or seven days). This dose was well tolerated. At day 3 or 7 after SCI, mice were euthanized with an overdose of Phenytoin/ Pentobarbital (120 mg/kg) and transcardially perfused with ice cold 1x PBS. A 5 mm piece of spinal cord centering on the lesion was dissected out, and immediately snap frozen. RNA was extracted using the RNeasy Lipid Tissue Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions, followed by RNA quantification and characterization of purity using Nanodrop (Thermo Scientific) and reverse transcription of 1 µg of RNA using QuantiTect Reverse Transcription Kit (Qiagen). Q-PCR was performed in duplicates using a PCR thermal cycler (LightCycler® 480 System, Roche) with specific primers using LightCycler® 480 Mastermix (Roche). Gene expression levels were analyzed using the ddct method normalized to peptidylprolyl isomerase A (PPIA) as a housekeeping gene and laminectomy as a baseline. The following primers were used for Q-PCR: PPIA: ATG TGC CAG GGT GGT GAC TTT A (forward primer 5'–3'); TGT GTT TGG TCC AGC ATT TGC C (reverse primer 5'–3'), TNF: TTG CTC TGT GAA GGG AAT GG (forward primer 5'–3'); GGC TCT GAG GAG TAG ACA ATA AAG (reverse primer 5'–3'), IL-1b: ATG GGC AAC CAC TTA CCT ATT T (forward primer 5'–3'); GTT CTA GAG AGT GCT GCC TAA TG (reverse primer 5'–3'), IL-6: CTT CCA TCC AGT TGC CTT CT (forward primer 5'–3'); CTC CGA CTT GTG AAG TGG TAT AG (reverse primer 5'–3'). iNOS: GAA CGG AGA ACG TTG GAT TTG (forward primer 5'–3'); TCA GGT CAC TTT GGT AGG ATT T (reverse primer 5'–3'), bax: CAT CTT CTT CCA GAT GGT GA (forward primer 5'–3'); GTT TCA TCC AGG ATC GAG CAG (reverse primer 5'–3').

DATA USAGE NOTES: Cohort 1: The average BMS score was significantly lower in gallium-treated mice at day 1 and 3. A lower number of gallium-treated mice were able to support their weight at day 1, 3 and 5 after SCI. Cohort 2: BMS scores and weight support at day 1 after injury were not different between groups. Q-PCR revealed no differences in expression levels of TNF, IL-6, IL-1ß, bax or iNOS between gallium maltolate and saline treated mice at day 3 and 7 after SCI. Bax, IL6 and iNOS showed a trend for higher expression levels in gallium-treated mice at day 3 after injury. Gallium treatment did not improve the functional outcome after SCI early after injury. However, the deterioration compared to controls could also have been caused by worse overall physiologic conditions of the mice. In the second cohort, no difference was seen between the groups at day 1. No anti-inflammatory effect was detected by Q-PCR at day 3 or 7. In summary, we did not detect any beneficial effect on functional outcome or Q-PCR level in gallium maltolate-treated mice.

Alteration of H-reflex and MEP amplitude in the flexor carpi radialis muscle with transcutaneous spinal cord stimulation during static and leg cycling tasks in neurologically intact male and female humans

DOI:10.34945/F5B59T

DATASET CITATION:

Parhizi B., Barss T. S., Mushahwar V. K. (2021) Alteration of H-reflex and MEP amplitude in the flexor carpi radialis muscle with transcutaneous spinal cord stimulation during static and leg cycling tasks in neurologically intact male and female humans. Open Data Commons for Spinal Cord Injury. ODC-SCI:602 http://dx.doi.org/10.34945/F5B59T

ABSTRACT:

STUDY PURPOSE: Transcutaneous spinal cord stimulation (tSCS) is a novel, non-invasive approach used to activate and modulate spinal networks. tSCS facilitates upper extremity function, generates locomotor-like responses, and may facilitate sensorimotor recovery after neural injury when paired with rehabilitation interventions. However, one drawback of tSCS is that its underlying mechanisms of action are not well understood. It has been suggested recently that tSCS modulates spinal circuitry in a manner similar to epidural spinal cord stimulation. In this study, we investigated the ability of tSCS applied to the lumbar and cervical spinal cord to modulate cervical circuitry. We raised two questions in this study: 1) To what extent can the cervical circuitry of the spinal cord be modulated by tSCS as measured in the flexor carpi radialis (FCR) by Hoffmann (H-)reflex and motor evoked potential (MEP) amplitude? 2) Does providing tSCS at multiple segments of the spinal cord (lumbar and cervical combined) converge to alter excitability of H-reflex or MEP amplitude to a greater extent compared to either site alone? The neuromodulatory effects of tSCS were assessed during both a static task and relative to a leg cycling paradigm to determine if tSCS alters cervico-lumbar coupling.

DATA COLLECTED: To explore the changes in amplitude of FCR H-reflexes and MEPs, 14 neurologically-intact participants completed the H-reflex (3 female, 11 male) and MEP (4 female, 10 male) assessments, with 11 completing both protocols. Because 3 individuals were excluded from MEP assessment due to possible contraindications to transcranial magnetic stimulation (TMS), 3 additional participants were recruited to complete only the MEP portion of the protocol. For each measure, participants completed two tasks that include a static task and leg cycling at a constant speed (?60 rpm). During each task, participants completed 4 tSCS conditions: (1) No tSCS, 2) tSCS active over the cervical spinal cord (Cervical); 3) tSCS active over the lumbar spinal cord (Lumbar); 4) tSCS active simultaneously on cervical and lumbar spinal cord (Combined). Thus, for each participant in each group 8 experimental conditions are available. Additionally, information about the level of background muscle activity, amplitude of evoked motor response (M-wave), maximum evoked motor response (Mmax), maximum MEP amplitude (MEPmax), peak muscle activation during a maximum voluntary contraction (MVC), and the amplitude of stimulation at the cervical and lumbar sites for each participant are included. The peak-to-peak amplitude of M-wave, H-reflex, and MEP, as well as baseline activity of the FCR muscle, were analyzed in a window of 400 ms (staring 100ms pre-stimulus, ending 300ms post-stimulus). A window of 100ms pre-stimulus (-100ms to 0ms relative to stimulus onset) was selected to calculate the baseline FCR and ECR EMG activity averaged over ten sweeps for each experimental condition. To obtain the value of pre-stimulus muscular contraction, the mean of the signal in this 100ms window was calculated and subtracted from the whole trace to remove any offset in the signal. The pre-stimulus background activity was then rectified and calculated as the mean activity in the 100ms window. The peak-to-peak amplitude of post-stimulus H-reflex, M-wave, and MEP were calculated by averaging ten sweeps per condition. The average values were then normalized to the value of Mmax for H-reflex measurements and to the value of MEPmax for MEP measurements, obtained in a separate trial immediately before the initiation of the testing conditions. The post-stimulus window of analysis for each evoked response was selected based on visual inspection. The FCR H-reflex was evoked by stimulating the median nerve using bipolar electrodes with 1ms square wave pulses. Transcranial magnetic stimulation was applied to the contralateral motor cortex using a double cone coil to elicit MEPs with a single monophasic pulse. Transcutaneous stimulation of the spinal cord was delivered by a constant current stimulator through two 2.5 cm round cathodic electrodes placed midline at C3-4 and C6-7, and T11 and L1 spinous processes. Two 5 × 10 cm rectangular electrodes were placed bilaterally over the iliac crests as anodes for the cervical tSCS while two additional anode electrodes were placed laterally for the lumbar tSCS. Muscle activity of four muscles in the left arm was recorded via electromyography (EMG) during each trial: FCR, extensor carpi radialis (ECR), biceps brachii (BB) and triceps brachii (TB). Muscle activity was recorded from surface Ag-AgCl electrodes placed on the muscle belly and recorded at a sampling rate of 2000Hz. All EMG signals were amplified 1000x during data collection and band-pass filtered from 30 to 1000 Hz. The EMG signals were used to record H-reflexes and MEPs from the FCR muscle. The amplitude of FCR H-reflexes, M-waves, and MEPs along with FCR/ECR pre-stimulus baseline activity were compared across different experimental conditions using repeated-measure ANOVA (rmANOVA). During the static task, the effects of condition (No tSCS, Cervical, Lumbar, and Combined) were compared for H-reflex, MEP, M-wave and baseline muscle activity with a 1 x 4 ANOVA. Similarly, during the cycling task, the effects of condition (Static No-tSCS, Cycle No-tSCS, Cycle Cervical, Cycle Lumbar, and Cycle Combined) were compared for H-reflex, MEP, M-wave, baseline muscle activity, and cycling cadence with a 1 x 5 ANOVA. Significant effects were followed by pairwise comparisons corrected by Tukey’s HSD adjustment for multiple comparisons. Differences with p ? 0.05 were accepted as statistically significant. This dataset represents a stand-alone data.

DATA USAGE NOTES: This study demonstrates, for the first time, that tonic activation of spinal cord networks through multiple sites of tSCS provides a facilitation of both spinal reflex and corticospinal pathways.

Hemodynamics, weight and overground locomotion from 1125 rats with different spinal cord injury thoracic contusion severities recovered from the Multicenter Spinal Cord Injury Study (MASCIS)

DOI:10.34945/F5QG66

DATASET CITATION:

Almeida C. A., Torres-Espin A., Huie J. Russell., Sun D., Noble-Haeusslein L. J., Young W., Beattie M. S., Bresnahan J. C., Nielson J. L., Ferguson A. R. (2021) Hemodynamics, weight and overground locomotion from 1125 rats with different spinal cord injury thoracic contusion severities recovered from the Multicenter Spinal Cord Injury Study (MASCIS). Open Data Commons for Spinal Cord Injury. ODC-SCI:595 http://dx.doi.org/10.34945/F5QG66

ABSTRACT:

STUDY PURPOSE: Results of the process of archeological data recovery of spinal cord injury (SCI) data sets from the Multicenter Spinal Cord Injury Study (MASCIS). The goal for this subset of the MASCIS dataset was to externally validate previous fundings on the importance of peri-surgery blood pressure in long term recovery.

DATA COLLECTED: This data represents a derivative dataset that involved large scale data integration and analysis of different epochs of the MASCIS study. MASCIS is multi-site preclinical drug trial conducted in the 1990s with the primary goal of testing the efficacy of the anti-inflammatory glucocorticoid methylprednisolone, and other therapies administered acutely after an SCI in rats. The dataset contains diverse data from 1125 rats. Different groups of injury were conducted using a thoracic contusion injury model. The presented subset of the data contains summaries of blood pressure, weight and BBB scores in different injury severity groups.

DATA USAGE NOTES: This dataset constitutes a demonstration of the value of data archeology. The curation of the MASCIS study has generated this data resource that can be use for secondary analysis and further discovery.

Acute post-injury blockade of a2d-1 calcium channel subunits on pathological autonomic plasticity after T3 crush spinal cord injury in female mice

DOI:10.34945/F5V30C

DATASET CITATION:

Brennan F. H.., Noble B. T.., Wang Y., Guan Z., Davis H., Mo X., Harris C., Eroglu C., Ferguson A. R.., Popovich P. G.. (2024) Acute post-injury blockade of a2d-1 calcium channel subunits on pathological autonomic plasticity after T3 crush spinal cord injury in female mice. Open Data Commons for Spinal Cord Injury. ODC-SCI:555 http://dx.doi.org/10.34945/F5V30C

ABSTRACT:

STUDY PURPOSE: After a spinal cord injury (SCI), normally innocuous visceral or somatic stimuli can trigger uncontrolled reflex activation of sympathetic circuitry, causing pathological dysautonomia. Here, we tested the hypothesis that SCI causes remarkable structural remodeling and synaptic plasticity, creating abnormal spinal sympathetic reflexes that promote dysautonomia. We also tested whether treating mice with prophylactic gabapentin (GBP), an FDA-approved drug that inhibits a2d-1-mediated synaptogenesis in the brain, can prevent these pathologies.

DATA COLLECTED: The dataset includes n = 183 adult female mice on a C57BL/6J background and n=16 adult female mice on a 129SVE background with a2d-1 overexpression or wild type littermates. Mice received a T3 crush SCI or sham control surgery. Mice were injected subcutaneously 3x/d with GBP (66.7 mg/kg) or Saline from 1 day post-injury (dpi) until 28 or 35 days post-injury, and were perfused at 28, 35, 42, or 56 dpi. Additional mice were used for pharmacokinetic studies and time-course studies of anatomical reorganization (sham, 7 dpi, 14 dpi, 21 dpi, 28 dpi). Anatomical reorganization was assessed using Vglut2+ pre-synaptic puncta, Vglut2+Homer1+ excitatory synapse count, quantification of spinal PRV+ neurons that control lymphoid tissue, lumbar CGRP fiber sprouting, and FosB+ neuron counts. Dysautonomia was assessed using spleen weight, spleen B and T cell quantification, and spontaneous and induced autonomic dysreflexia experiments. Additional outcome measures include a2d-1+ cell number, percent a2d-1+ area, thrombospondin mRNA and protein levels, and principal component analysis results (spinal sympathetic reflex index).

CONCLUSIONS:

Minocycline treatment for acute cervical spinal cord injury in female rats: behavioural, histopathological and systemic immune effects

DOI:10.34945/F5JS3M

DATASET CITATION:

Schmidt E. K A., Raposo P. J F., Torres-Espin A., Fenrich K. K., Fouad K. (2021) Minocycline treatment for acute cervical spinal cord injury in female rats: behavioural, histopathological and systemic immune effects. Open Data Commons for Spinal Cord Injury. ODC-SCI: 457 http://dx.doi.org/10.34945/F5JS3M

ABSTRACT:

STUDY PURPOSE: Minocycline has been widely studied for its local anti-inflammatory properties; however little is known about its antibiotic and systemic immune effects following spinal cord injury (SCI). The aim of the present study is to elucidate multiple system-wide changes involving the microbiota-immune axis induced my minocycline treatment in a rodent model of cervical contusion SCI (unilateral contusion 125 kdyns at an angle of 15 degrees to the right at C5). This study included 4 groups of adult female Lewis rats: uninjured n=10, uninjured + minocycline n=10, SCI n=8, SCI + minocycline n=9. Minocycline (or sterile water for untreated control groups) was administered at a dose of 50mg/kg via oral gavage for 7 days immediately following injury..

DATA COLLECTED: pICRUST bioinformatics software was used to predict the metagenomic functional content of the 16s rRNA data; behavioural data from the open field and cylinder tasks, anxiety- and depressive-like behaviour was analyzed in the elevated plus maze, light/dark box and sucrose preference tests; quantification of the lesion area and extension; 29 plasma analytes (cytokines, chemokines and hormones) were measured before injury, 5, 14, and 28 days post injury; quantification of microglia (iba1) immunoreactivity above, at and below the lesion site; body weight monitored weekly throughout the experiment. 16s rRNA sequencing data from fecal samples collected before injury, on the day of injury, 5, 14 and 28 days post injury is located in a separate dataset.

DATA USAGE NOTES: The present data shows that 7 days of minocycline treatment had a drastic acute effect on the microbiota composition regardless of whether the animal was uninjured or had a SCI. SCI resulted in a delayed (measured 28 days after injury) suppression of inflammatory cytokines/chemokines in the plasma, which was prevented with minocycline treatment. Minocycline treatment had a modest but insignificant anxiolytic effect in the light/dark box following SCI. Finally, we did not find minocycline treatment to attenuate lesion size or extension, however there was an increase in iba1 immunoreactivity above and below the lesion site in the minocycline + SCI group compared to untreated SCI rats.

Acute blood RNA sequencing from SCI human patients

DOI:10.34945/F5QC7J

DATASET CITATION:

Kyritsis N., Torres Espin A., Schupp P. G., Huie J. Russell., Chou A., Duong-Fernandez X., Thomas L. H., Tsolinas R. E., Hemmerle D. D., Pascual L. U., Singh V., Pan J. Z., Talbott J. F., Whetstone W. D., Burke J. F., DiGiorgio A. M., Weinstein P. R., Manley G. T., Dhall S. S., Ferguson A. R., Oldham M. C., Bresnahan J. C., Beattie M. S. (2021) Acute blood RNA sequencing from SCI human patients. Open Data Commons for Spinal Cord Injury. ODC-SCI: 405 http://dx.doi.org/10.34945/F5QC7J

ABSTRACT:

STUDY PURPOSE: The purpose of this study is to discover, through high throughput methods, blood RNA biomarkers with the ability to diagnose accurately the severity of the spinal cord injury at the acute stage.

DATA COLLECTED: Total RNA was collected from white blood cells of 38 spinal cord injury patients, 10 healthy subjects and 10 non-CNS trauma patients. The samples were collected on average within the first 24 hours after injury. Subsequently the RNA was used for RNAseq. These subjects were enrolled in the Transforming Research and Clinical Knowledge in Spinal Cord Injury (TRACK-SCI) clinical study, and we included clinical variables such as age, sex, race, prior CNS pathology, concurrent TBI, injury severity score, AIS grade, neurological level of injury, and motor and sensory scores. Both clinical and transcriptomic data were used to generate a predictive model for the initial severity of spinal cord injury.

DATA USAGE NOTES: Analysis of our gene expression data revealed 197 genes that they are expressed in an severity-dependent manner. Furthermore, unsupervised gene co-expression network analysis was used for the discovery of 16 gene modules that are induced upon spinal cord injury. A logistic regression model using these modules displayed high accuracy in diagnosing the initial injury severity.

Data for the manuscript: Fecal Transplant Prevents Gut Dysbiosis and Anxiety-like Behaviour After Spinal Cord Injury in Rats

DOI:10.7295/W97942VQ

DATASET CITATION:

Schmidt, E., Torres-Espin, A., Raposo, P., Madsen, K., Kigerl, K., Popovich., P., Fenrich, K.F., Fouad, K. (2020) Data for Fecal Transplant Prevents Gut Dysbiosis and Anxiety-like Behaviour After Spinal Cord Injury in Rats. Open Data Commons for Spinal Cord Injury. ODC-SCI:262 http://doi.org/10.7295/W97942VQ

ABSTRACT:

STUDY PURPOSE: To establish a model of anxiety following a cervical contusion spinal cord injury (SCI) in rats and to determine whether the microbiota play a role in the observed behavioural changes.

DATA COLLECTED: This dataset includes n=57 rats from 2 experiments. Experiment 1: sham (underwent surgery with no SCI) n=6; unilateral cervical contusion SCI n=6. Experiment 2: Healthy (no operation, no gavage) n=10; Sham n=11; SCI (gavaged with a control solution) n=10; SCI-FMT (gavaged with fecal microbiota transplant solution) n=14. A subset of subjects from experiment 2 were used for the fecal 16s rRNA analysis (healthy n = 10; SCI-FMT n = 10; sham n = 5; SCI n = 5). Fecal matter for the 16s rRNA analysis were collected before injury, 3 days after injury and 4 weeks after injury. A PICRUST analysis was performed to infer the functional pathways involved using the 16s rRNA gene data. Rats were assessed on a battery of behavioural tests: the light-dark box, the cylinder test, the sucrose preference test, the elevated plus maze and the open field. Lesion size was calculated as the percentage of damaged tissue area throughout the rostral-caudal extension of the injury site.

PRIMARY CONCLUSION: Treatment with a fecal microbiota transplant in the acute post-injury period prevents spinal cord injury-induced gut dysbiosis as well as the development of anxiety-like behaviour.

Repeated measurements of hindlimb CatWalk variables in normal rats

DOI: 10.34945/F54S3W

DATASET CITATION:

Aceves, M., Dietz, V. A., Dulin, J. N., Jeffery, U., Jeffery, N. D. 2020. Repeated measurements of hindlimb CatWalk variables in normal rats. Open Data Commons for Spinal Cord Injury. ODC-SCI:432. doi: 10.34945/F54S3W

ABSTRACT:

STUDY PURPOSE: To provide a set of reference data for expected variability in hindlimb locomotor outcomes for normal young adult rats using the Catwalk.

DATA COLLECTED: Data that are automatically collected on hindlimb function in rats by the CatWalk. Recordings were made on 16 male 9-week-old (250-275 g) Sprague-Dawley rats after a training period and at 8 weekly intervals.

PRIMARY CONCLUSION: There is a large variation in within-rat variability between different CatWalk outcomes. Selection of the most appropriate variables can improve sensitivity of experiments that measure locomotor outcome.

T10 lateral hemisection spinal cord injury with multiple histological and behavioral outcomes

DOI:10.7295/W9HQ3X20

DATASET CITATION:

Liu, Y., Wang, X., Li, W., Zhang, Q., Li, Y., Zhang, Z., Zhu, J., Chen, B., Williams, P.R., Zhang, Y., Yu, B., Gu, X., He, Z. (2019). T10 lateral hemisection spinal cord injury with multiple histological and behavioral outcomes. Open Data Commons for Spinal Cord Injury. ODC-SCI:212 http://doi.org/10.7295/W9HQ3X20

ABSTRACT:

STUDY PURPOSE: A major hurdle for functional recovery after spinal cord injury is the limited re- growth of the axons in the corticospinal tract (CST) that originate in the motor cortex and innervate the spinal cord. In this study, we tested whether post-lesional AAV-assisted co- expression of two soluble proteins, namely insulin-like growth factor 1 (IGF1) and osteopontin (OPN), in cortical neurons leads to CST regrowth and CST-dependent functional recovery.

DATA COLLECTED: With an incomplete spinal cord injury model (T10 lateral hemisection), we tested the sensitizing effect of OPN/IGF1 on corticospinal neurons, the extent of axon regrowth both in the injured side (axon regeneration) and the intact side (collateral axon sprouting). In addition, we assessed behavioral performance for both gross and skilled locomotion in mice treated with OPN/IGF1 post T10 lateral hemisection.

PRIMARY CONCLUSION: Our results demonstrate a potentially translatable strategy for restoring cortical dependent function after injury in the adult.