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://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.

KEYWORDS

spinal gray matter; Spinal Cord Injury; spinal interneurons; behavioral deficits; lumbar enlargement; spontaneous functional recovery

PROVENANCE / ORIGINATING PUBLICATIONS

• Intermediate Gray Matter Interneurons in the Lumbar Spinal Cord Play a Critical and Necessary Role in Coordinated Locomotion Naëmi Kuehn, Andreas Schwarz, Carlo Antonio Beretta, Yvonne Schwarte, Francesca Schmitt, Melanie Motsch, Norbert Weidner, Radhika Puttagunta bioRxiv 2022.10.31.514612; doi: https://doi.org/10.1101/2022.10.31.514612. .

RELEVANT LINKS

NOTES

DATASET INFO

Contact: Puttagunta Radhika (radhika.puttagunta@med.uni-heideleberg.de)

Lab: Puttagunta Lab

ODC-SCI Accession:938

Records in Dataset: 145

Fields per Record: 49

Last updated: 2023-08-31

Date published: 2023-08-31

Downloads: 5

Files: 2

LICENSE

Creative Commons Attribution License (CC-BY 4.0)

FUNDING AND ACKNOWLEDGEMENTS

Deutsche Forschungsgemeinschaft grants SFB 1158 A06 (RP and NW), PU 425/4-3 (RP), WE 2165/4-3 (NW)

CONTRIBUTORS

Kuehn, Naëmi

Laboratory for Experimental Neuroregeneration, Spinal Cord Injury Center, Heidelberg University Hospital, Heidelberg, Germany

Schwarz, Andreas

Laboratory for Experimental Neurorehabilitation, Spinal Cord Injury Center, Heidelberg University Hospital, Heidelberg, Germany

Beretta, Carlo A.

Department of Functional Neuroanatomy, Institute for Anatomy and Cell Biology, Heidelberg University, Heidelberg, Germany

Schwarte, Yvonne

Laboratory for Experimental Neuroregeneration, Spinal Cord Injury Center, Heidelberg University Hospital, Heidelberg, Germany

Schmitt, Francesca

Laboratory for Experimental Neuroregeneration, Spinal Cord Injury Center, Heidelberg University Hospital, Heidelberg, Germany

Motsch, Melanie

Laboratory for Experimental Neuroregeneration, Spinal Cord Injury Center, Heidelberg University Hospital, Heidelberg, Germany

Weidner, Norbert

Spinal Cord Injury Center, Heidelberg University Hospital, Heidelberg, Germany

Puttagunta, Radhika [ORCID:0000-0001-7674-8064]

Laboratory for Experimental Neuroregeneration, Spinal Cord Injury Center, Heidelberg University Hospital, Heidelberg, Germany