Gait Analysis of Adult Paraplegic Rats after Spinal Cord Repair

doi:10.1006/exnr.1997.6708

Experimental Neurology

Volume 148, Issue 2, December 1997, Pages 544-557

https://doi.org/10.1006/exnr.1997.6708 Get rights and content

Abstract

This study presents a novel detailed method of analysis of rat gait and uses this method to demonstrate recovery of forward locomotion patterns in adult rats made paraplegic by surgical spinal cord transection and subjected to a novel strategy for spinal cord repair. Six normal rats were compared to five animals in which the cord was transected at T8–T9, and a 5-mm segment of the spinal cord removed, and to seven animals in which, following spinal cord transection and removal of a spinal cord segment, multiple intercostal peripheral nerve bridges were implanted, rerouting pathways from white to gray matter in both directions. The implanted area was filled with fibrin glue containing acidic fibroblast growth factor. Details of the repair strategy have been published (H. Cheng, Y. Cao, and L. Olson, 1996,Science273:510–513). Gait analysis was carried out 3 and 4 months after surgery and once in the normal animals. Animals were allowed to walk across a runway with a transparent floor. Each test consisted of five trials, and each trial was videorecorded from underneath. Using frame-by-frame playback, individual footprints were then recorded regarding location and order of limb use, as well as step quality (degree of weight bearing, etc.). These data allowed measuring runway transit time, five different measures of step numbers, all possible temporal patterns of limb use, stride length, and base of support. Transected controls remained paralyzed in the hindlimbs with only occasional reflex hindlimb movements without weight bearing. Animals subjected to the full repair procedure were significantly faster than the controls, used their hindlimbs for 25–30% of the movements, and regained several of the specific limb recruitment patterns used by normal rats. Taken together, the gait analysis data demonstrate remarkable recovery of coordinated gait in the repaired animals, which was significantly better than controls for all relevant parameters, while at the same time clearly inferior to normal rats for most of the examined parameters. We conclude that normal rats use a multitude of interchangeable step sequence patterns, and that our spinal cord repair strategy leads to recovery of some of these patterns following complete spinal cord transection. These data suggest functionally relevant neuronal communication across the lesion.

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Edema formation is one of the very first events to occur after spinal cord injury (SCI) leading to an increase of the intrathecal pressure and consequently to serious spinal tissue and functional impairments. Current edema treatments are still symptomatic and/or non-specific. Since edema formation mechanisms are mainly described as vasogenic and cytotoxic, it becomes crucial to understand the interplay between these two subtypes. Acting on key targets to inhibit edema formation may reduce secondary damage and related functional impairments. In this study, we characterize the edema kinetic after T9–10 spinal contusion. We use trifluoperazine (TFP) to block the expression and the functional subcellular localization of aquaporin-4 supposed to be implicated in the cytotoxic edema formation. We also use sodium cromoglycate (SCG) to deactivate mast cell degranulation known to be implicated in the vasogenic edema formation. Our results show a significant reduction of edema after TFP treatment and after TFP-SCG combined treatment compared to control. This reduction is correlated with limited onset of initial sensorimotor impairments particularly after combined treatment. Our results highlight the importance of potential synergetic targets in early edema therapy after SCI as part of tissue sparing strategies.
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Spinal cord injury (SCI) is a devastating disorder that often results in severe sensorimotor function impairment with limited recovery of function. In recent years, rehabilitation training for spinal cord injury has gradually emerged, and some of them play an important role in the repair of spinal cord injury However, the optimal training regimen for SCI remains to be determined. In this study, we explore the effects of rotarod training (began at 7 days post-injury) on the recovery of motor function after SCI, as well as its possible repair mechanism from the aspects of function and histopathological changes, the behaviors of specific trophic factors and cytokines, and the expression profile of specific genes. Multiple functional assessments showed that rotarod training initiated at 7 days post-injury is unsuitable for promoting neuro-electrophysiological improvement and trunk stability, but impaired functional coordination and motor recovery. In addition, rotarod training has negative effects on spinal cord repair after SCI, which is manifested as an increase of lesion area, a decrease in neuronal viability, a deterioration in immuno-microenvironment and remyelination, a significant reduction in the expression of trophic factors and an increase in the expression of pro-inflammatory factors. RNA sequencing suggested that the genes associated with angiogenesis and synaptogenesis were significantly downregulated and the PI3K-AKT pathway was inhibited, which was detrimental to spinal cord repair and impeded nerve regeneration. These results indicate that immediate rotarod training after SCI is currently unsuitable for rehabilitation in mice.
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Step Sequence, which refers to the Normal Step Sequence Patterns (NSSP) and the Regularity Index (RI). The NSSP (Fig. 1) are six different sequences of paw placements (PP), which can be observed in healthy animals and vary between different strains of rats as well different speed velocities (Cheng et al., 1997; Koopmans et al., 2007a). The RI expresses the amount of NSSP in correlation to the total amount of paw placements during one CW crossing: RI = (NSSP × 4/PP) × 100 %.
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The central nervous system (CNS) plays a pivotal role in virtually all mammalian functions, including cognitive behavior, motility, perception, and homeostasis/regulation. Trauma to the CNS can therefore have devastating consequences due to the disruption of complex neural circuitry underlying these critical functions, which is compounded by the limited regenerative capacity of the brain and spinal cord. Rat models of both spinal cord injury and traumatic brain injury are described in this chapter. Induction of specific models and selected outcome measures are reviewed. Benefits of using rat models, potential confounding factors on animal experiments, and complications for interpretation of data obtained from these experiments are highlighted.
Changes in innervation of lumbar motoneurons and organization of premotor network following training of transected adult rats
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Stepping pattern: stepping patterns were classified as normal when the animal placed the four paws one after another in an alternate, cruciate or rotate pattern. Cruciate or rotate refer diagonal and lateral sequences (Gillis and Biewener, 2001), respectively, in the order of limb recruitment (Cheng et al., 1997). Adult rats were anaesthetized by i.p. administration of a mixture of ketamine (Imalgene, Merial, 60 mg/k) and xylazine (Rompun, Bayer, 10 mg/kg).
Rats with complete spinal cord transection (SCT) can recover hindlimb locomotor function under strategies combining exercise training and 5-HT agonist treatment. This recovery is expected to result from structural and functional re-organization within the spinal cord below the lesion. To begin to understand the nature of this reorganization, we examined synaptic changes to identified gastrocnemius (GS) or tibialis anterior (TA) motoneurons (MNs) in SCT rats after a schedule of early exercise training and delayed 5-HT agonist treatment. In addition, we analyzed changes in distribution and number of lumbar interneurons (INs) presynaptic to GS MNs using retrograde transneuronal transport of rabies virus.
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^☆: M. BrownM. GoldergerB. Bregman, Eds.

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Regular ArticleGait Analysis of Adult Paraplegic Rats after Spinal Cord Repair☆

Abstract

Prog. Neurobiol.

Brain Res.

Exp. Neurol.

Exp. Neurol.

Exp. Neurol.

Brain Res.

Prog. Brain Res.

Neuroscience

Curr. Opin. Neurobiol.

Exp. Neurol.

Brain Res.

Brain Res.

Brain Res.

Brain Res.

Exp. Neurol.

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Exp. Neurol.

A sensitive and reliable locomotor rating scale for open field testing in rats

J. Neurotrauma

Longitudinal assessment of locomotor recovery in rats with spinal cord impact lesions

Criteria for the Assessment of Recovery of Function: Behavioral Methods

Spinal cord injury produced by consistent mechanical displacement of the cord in rats: Behavioral and histologic analysis

J. Neurotrauma

Kinematic analysis of limb position during quadrupedal locomotion in rats

J. Neurotrauma

The intrinsic factors in the act of progression in the mammals

Proc. R. Soc. London

Fibrin glue used as an adhesive agent in CNS tissues

J. Neural Transplant. Plast.

Spinal cord repair in adult paraplegic rats: Partial restoration of hindlimb function

Science

Regular Article
Gait Analysis of Adult Paraplegic Rats after Spinal Cord Repair☆