Selective plasticity of primary afferent innervation to the dorsal horn and autonomic nuclei following lumbosacral ventral root avulsion and reimplantation in long term studies

Previous studies involving injuries to the nerves of the cauda equina and the conus medullaris have shown that lumbosacral ventral root avulsion in rat models results in denervation and dysfunction of the lower urinary tract, retrograde and progressive cell death of the axotomized motor and parasympathetic neurons, as well as the emergence of neuropathic pain. Root reimplantation has also been shown to ameliorate several of these responses, but experiments thus far have been limited to studying the effects of lesion and reimplantation local to the lumbosacral region. Here, we have expanded the region of investigation after lumbosacral ventral root avulsion and reimplantation to include the thoracolumbar sympathetic region of the spinal cord. Using a retrograde tracer injected into the major pelvic ganglion, we were able to define the levels of the spinal cord that contain sympathetic preganglionic neurons innervating the lower urinary tract. We have conducted studies on the effects of the lumbosacral ventral root avulsion and reimplantation models on the afferent innervation of the dorsal horn and autonomic nuclei at both thoracolumbar and lumbosacral levels through immunohistochemistry for the markers calcitonin gene-related peptide (CGRP) and vesicular glutamate transporter 1 (VGLUT1). Surprisingly, our experiments reveal a selective and significant decrease of CGRP-positive innervation in the dorsal horn at thoracolumbar levels that is partially restored with root reimplantation. However, no similar changes were detected at the lumbosacral levels despite the injury and repair targeting efferent neurons, and being performed at the lumbosacral levels. Despite the changes evident in the thoracolumbar dorsal horn, we find no changes in afferent innervation of the autonomic nuclei at either sympathetic or parasympathetic segmental levels by CGRP or VGLUT1. We conclude that even remote, efferent root injuries and repair procedures can have an effect on remote and non-lesioned sensory systems sharing common peripheral ganglia.     

Surgical implantation of avulsed lumbosacral ventral roots promotes restoration of bladder morphology in rats

Injuries to the cauda equina and conus medullaris of the spinal cord commonly result in paraplegia, sensory deficits, neuropathic pain, as well as bladder, bowel, and reproductive dysfunctions. In a recently developed lower motoneuron model for cauda equina injury and repair, we have demonstrated that an acute surgical implantation of avulsed lumbosacral ventral roots into the conus medullaris is neuroprotective, promotes regeneration of efferent spinal cord axons into the implanted roots, and may result in functional reinnervation of the lower urinary tract. Here, we investigated the effects of a bilateral lumbosacral ventral root avulsion (VRA) injury and re-implantation on the morphology of the rat bladder at twelve weeks post-operatively. We demonstrated a VRA-induced overall thinning of the bladder wall, which exhibited reduced thickness of both the lamina propria and smooth muscle. In contrast, the bladder epithelium markedly increased its thickness in the injured series. Quantitative immunohistochemical studies showed a selective increase in CGRP immunoreactivity in the lamina propria after the VRA injury. Interestingly, the injury-induced changes in bladder wall morphology were ameliorated by an acute implantation of the lesioned roots into the conus medullaris. Specifically, bladders of the implanted group showed a partial restoration of the thickness of the lamina propria and epithelium as well as a return of CGRP immunoreactivity to baseline levels in the lamina propria. Our results support the notion that surgical implantation of severed ventral roots into the spinal cord may promote the recovery of a normal morphological phenotype in peripheral end organs.     

Do synaptic rearrangements underlie compensatory reflex enhancement in spinal motoneurons after partial cell loss?

In adult cats, avulsion of a spinal ventral root induces retrograde cell death among the corresponding motoneurons and, also, enhanced monosynaptic reflexes ipsilaterally in the adjacent uninjured spinal cord segments. The present study investigates possible mechanisms behind this reflex potentiation. At 1-12 weeks after unilateral L7 ventral root avulsion, the L7 dorsal root ganglia were bilaterally injected with choleragenoid-HRP to light microscopically quantify the amount of HRP-labeled terminals in the motor nuclei of the lesioned L7 segment and adjacent intact L6+S1 segments. In addition, motoneuron synaptology and individual HRP-labeled boutons were analyzed electron microscopically. In the L7 segment, the loss of motoneurons at 12 weeks after ventral root avulsion was accompanied by a marked loss of HRP-labeled boutons in the corresponding ventral horn. In the L6/S1 segments, the monosynaptic reflex enhancement found ipsilaterally at 12 weeks postoperatively (mean 212%) was not accompanied by an increased HRP-labeling in the ventral horn (mean 109%), indicating that no sprouting or enlargement of the monosynaptic boutons had occurred. Ultrastructurally, the values for apposition length, total active site length, cross-sectional area, and mitochondrial density of the labeled boutons were also similar between the two sides. However, ipsilaterally the L6/S1 motoneurons exhibited an increased membrane covering by presumably excitatory boutons. The present results indicate that after partial cell death in a motoneuron pool the remaining motoneurons may undergo compensatory synaptic rearrangements leading to increased excitability and enhanced reflexes.     

At-level neuropathic pain is induced by lumbosacral ventral root avulsion injury and ameliorated by root reimplantation into the spinal cord

Neuropathic pain is common after traumatic injuries to the cauda equina/conus medullaris and brachial plexus. Clinically, this pain is difficult to treat and its mechanisms are not well understood. Lesions to the ventral roots are common in these injuries, but are rarely considered as potential contributors to pain. We examined whether a unilateral L6-S1 ventral root avulsion (VRA) injury in adult female rats might elicit pain within the dermatome projecting to the adjacent, uninjured L5 spinal segment. Additionally, a subset of subjects had the avulsed L6-S1 ventral roots reimplanted (VRA+Imp) into the lateral funiculus post-avulsion to determine whether this neural repair strategy elicits or ameliorates pain. Behavioral tests for mechanical allodynia and hyperalgesia were performed weekly over 7 weeks post-injury at the hindpaw plantar surface. Allodynia developed early and persisted post-VRA, whereas VRA+Imp rats exhibited allodynia only at 1 week post-operatively. Hyperalgesia was not observed at any time in any experimental group. Quantitative immunohistochemistry showed increased levels of inflammatory markers in laminae III-V and in the dorsal funiculus of the L5 spinal cord of VRA, but not VRA+Imp rats, specific to areas that receive projections from mechanoreceptive, but not nociceptive, primary afferents. These data suggest that sustained at-level neuropathic pain can develop following a pure motor lesion, whereas the pain may be ameliorated by acute root reimplantation. We believe that our findings are of translational research interest, as root implantation surgery is emerging as a potentially useful strategy for the repair of cauda equina/conus medullaris injuries.     

Regeneration after spinal nerve root injury

Spinal nerve root avulsion has been considered as a central nervous type of injury and therefore not repaired surgically in man. The possibility for axonal regeneration after root avulsion or root lesion has been investigated in laboratory animals by means of up to date neurophysiological, morphological and tracing techniques. It is shown that, after ventral root avulsion and implantation into the spinal cord, alpha and probably also gamma motoneurons are able to regenerate within the spinal cord for a considerable distance before entering the implanted root and reinnervate previously denervated skeletal muscles. The regenerated neurons were found to respond to afferent activity with excitatory or inhibitory responses, and the regenerated axons could conduct action potentials that elicited muscle twitch responses. After dorsal root injury in the adult animal, regeneration into the spinal cord does not occur. However, regeneration of primary sensory neurons into appropriate locations of the spinal cord can be demonstrated in immature animals.     

Surgical reconstruction of spinal cord circuit provides functional return in humans

This mini review describes the current surgical strategy for restoring function after traumatic spinal nerve root avulsion in brachial or lumbosacral plexus injury in man. As this lesion is a spinal cord or central nervous injury functional return depends on spinal cord nerve cell growth within the central nervous system. Basic science, clinical research and human application has demonstrated good and useful motor function after ventral root avulsion followed by spinal cord reimplantation. Recently, sensory return could be demonstrated following spinal cord surgery bypassing the injured primary sensory neuron. Experimental data showed that most of the recovery depended on new growth reinnervating peripheral receptors. Restored sensory function and the return of spinal reflex was demonstrated by electrophysiology and functional magnetic resonance imaging of human cortex. This spinal cord surgery is a unique treatment of central nervous system injury resulting in useful functional return. Further improvements will not depend on surgical improvements. Adjuvant therapy aiming at ameliorating the activity in retinoic acid elements in dorsal root ganglion neurons could be a new therapeutic avenue in restoring spinal cord circuits after nerve root avulsion injury.     

Rescue and sprouting of motoneurons following ventral root avulsion and reimplantation combined with intraspinal adeno-associated viral vector-mediated expression of glial cell line-derived neurotrophic factor or brain-derived neurotrophic factor

Following avulsion of a spinal ventral root, motoneurons that project through the avulsed root are axotomized. Avulsion between, for example, L2 and L6 leads to denervation of hind limb muscles. Reimplantation of an avulsed root directed to the motoneuron pool resulted in re-ingrowth of some motor axons. However, most motoneurons display retrograde atrophy and subsequently die. Two neurotrophic factors, glial cell line-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF), promote the survival of motoneurons after injury. The long-term delivery of these neurotrophic factors to the motoneurons in the ventral horn of the spinal cord is problematic. One strategy to improve the outcome of the neurosurgical reinsertion of the ventral root following avulsion would involve gene transfer with adeno-associated viral (AAV) vectors encoding these neurotrophic factors near the denervated motoneuron pool. Here, we show that AAV-mediated overexpression of GDNF and BDNF in the spinal cord persisted for at least 16 weeks. At both 1 and 4 months post-lesion AAV-BDNF- and -GDNF-treated animals showed an increased survival of motoneurons, the effect being more prominent at 1 month. AAV vector-mediated overexpression of neurotrophins also promoted the formation of a network of motoneuron fibers in the ventral horn at the avulsed side, but motoneurons failed to extent axons into the reinserted L4 root towards the sciatic nerve nor to improve functional recovery of the hind limbs. This suggests that high levels of neurotrophic factors in the ventral horn promote sprouting, but prevent directional growth of axons of a higher number of surviving motoneurons into the implanted root.     

脊髓背根入髓区毁损术治疗脊髓和马尾神经损伤后疼痛的长期疗效分析

目的探讨脊髓背根入髓区（dorsal not entry zone,DREZ）显微外科毁损术对脊髓和马尾神经损伤后神经病理性疼痛的长期疗效和安全性。方法脊髓和马尾神经损伤后神经病理性疼痛35例,均行DREZ显微外科毁损术。对所有病人进行术前和术后视觉模拟疼痛评分（VAS）,以术后疼痛缓解〉75％为疗效优秀,疼痛缓解50％～75％为良好,疼痛缓解〈50％为差。结果术后2周疗效优秀33例（94．3％）,疗效差2例（5．7％）。长期随访中,疗效优秀24例（68．6％）,疗效良好6例（17．1％）,疗效差5例（14．3％）。结论DREZ显微外科毁损术对脊髓和马尾神经损伤后神经病理性疼痛长期疗效满意,并发症少,可明显提高病人的生活质量。     

Survival, regeneration and functional recovery of motoneurons after delayed reimplantation of avulsed spinal root in adult rat

We have established that extensive reinnervation and functional recovery follow immediate reimplantation of avulsed ventral roots in adult rats. In the present study, we examined the consequences of reimplantation delayed for 2 weeks after avulsion of the C6 spinal root. Twelve and 20 weeks after delayed reimplantation, 57% and 53% of the motoneurons in the injured spinal segment survived. More than 80% of surviving motoneurons regenerated axons into the reimplanted spinal root. Cholinesterase-silver staining revealed axon terminals on endplates in the denervated muscles. The biceps muscles in reimplanted animals had atrophied less than those in animals with avulsion only, as indicated by muscle wet weight and histological appearance. After electrical stimulation of the motor cortex or the C6 spinal root, typical EMG signals were recorded in biceps of reimplanted animals. The latency of the muscle potential at 20 weeks was similar to that of sham-operated controls. Behavioral recovery was demonstrated by a grooming test and ipsilateral forepaw movements were well coordinated in both voluntary and automatic activities. These results demonstrate that ventral root reimplantation can protect severed motoneurons, enable the severed motoneurons to regenerate axons, and enhance the recovery of forelimb function even when it is delayed for 2 weeks after avulsion.     

Delayed myeloradiculopathy produced by spinal X-irradiation in the rat

Rats were subjected to 3,500 r of X-irradiation in a single dose while breathing oxygen at 1 atm pressure. Comparison was made between the delayed effects of irradiating thoracic, lumbar, and the cauda equina fields. The lumbar field involved the alpha-motoneurons and spinal roots supplying the sciatic nerve, while the cauda equina field involved these spinal roots but spared the alpha-motoneurons in the spinal cord.Thoracic irradiation produced paraplegia after an interval of 127–150 days. In the irradiated zone, the spinal cord was severely damaged, but the thoracic spinal roots were spared.Lumbar irradiation produced paraplegia after an interval of 83–211 days. In the irradiated zone, the alpha-motoneurons were largely spared, the spinal cord showed mild to moderate white matter damage, but the most severe damage was of the lumbosacral spinal roots. The posterior roots were more affected than the anterior. In longer interval cases the degeneration of the roots appeared to be due to focal devitalization. Evidence is advanced that root degeneration had been progressing for at least 4 weeks before the onset of paraplegia.In the cauda equina series the lumbosacral spinal root changes were similar to those in the lumbar series.This study indicates that different levels of the neuraxis have different degrees of susceptibility to X-irradiation. The thoracic cord appears more susceptible than the lumbosacral; the lumbosacral roots appear more susceptible than the thoracic; the posterior roots are more susceptible than the anterior. These findings may have relevance to the study of radiation damage in man, even though the dose schedule used in this experimental study differs greatly from that used for radiotherapy.     

Improved detection of fluorogold-labeled neurons in long-term studies

Fluorogold (FG) is a widely used neuroanatomical tracer. However, because FG-labeled neurons become undetectable over time, its use has been limited in long-term studies. We investigated whether the detection of FG in retrogradely labeled neurons in long-term studies can be improved by immunohistochemistry (IHC) using an antibody to FG. We performed intraperitoneal injections of a FG solution to retrogradely label all parasympathetic preganglionic neurons (PPNs) and motoneurons (MNs) in the S1 spinal cord segment in adult rats. At 1, 6, and 12 weeks after the tracer injection, sections were immunohistochemically processed for FG and choline acetyltransferase (ChAT), an endogenous marker for all PPNs and MNs. Stereological counts demonstrated no cell loss of FG-labeled PPNs and MNs at 6 and 12 weeks. Cell size measurements showed that FG-immunolabeled neurons were smaller at 12 weeks, but not at 6 weeks. However, it is likely that there was no neuronal atrophy, but loss/degradation of the dye at a timepoint between 6 and 12 weeks, as ChAT-immunolabeled neurons showed no cell size reduction at 12 weeks. Our results suggest that the use of an antibody against FG improves the detection of FG for reliable neuronal counts and that the dye is not toxic to the retrogradely labeled neurons. We conclude that FG-labeling is a useful tool to determine neuronal counts in long-term studies, but should be used cautiously for neuronal size measurements.     

Anatomical evidence for two spinal 'afferent-interneuron-efferent' reflex pathways involved in micturition in the rat: a 'pelvic nerve' reflex pathway and a 'sacrolumbar intersegmental' reflex pathway

We labeled interneurons in the L1-L2 and L6-S1 spinal cord segments of the rat that are involved in bladder innervation using transneuronal retrograde transport of pseudorabies virus (PRV) in normal animals and in animals with selected nerve transections. Preganglionic neurons were identified using antisera against choline acetyltransferase (ChAT). In some experiments we labelled parasympathetic preganglionic neurons (PPNs) in the L6-S1 spinal cord by retrograde transport of Fluorogold from the major pelvic ganglion. We identified bladder afferent terminals using the transganglionic transport of the anterograde tracer cholera toxin subunit b. We present anatomical evidence for two spinal pathways involved in innervation of the bladder. First, in the intact rat, afferent information from the bladder connects, via interneurons in L6-S1, to the PPNs that provide the efferent innervation of the bladder. The afferent terminals were located mainly in close apposition to interneurons located dorsal to the retrogradely labeled PPNs. Second, using L6-S1 ganglionectomies or L6-S1 ventral root rhizotomies we limited viral transport to the sympathetic pathways innervating the bladder. This procedure also labelled interneurons (but not PPNs) with PRV in the L6-S1 spinal cord in a location very similar to those described in the intact rat. These interneurons also receive bladder afferent terminals but we propose that they project to sympathetic preganglionic neurons, most of which are in the L1-L2 spinal segments. Based on this anatomical evidence, we propose the existence of two spinal reflex pathways involved in micturition: a pathway limited to a reflex arc in the pelvic nerve (presumably excitatory to the detrusor muscle); and a pathway involving the pelvic nerve and sympathetic nerve fibers, some of which may travel in the hypogastric (presumably inhibitory to the detrusor muscle).     

Neuroprotection and Axonal Regeneration After Lumbar Ventral Root Avulsion by Re-implantation and Mesenchymal Stem Cells Transplant Combined Therapy

Ventral spinal root avulsion causes complete denervation of muscles in the limb and also progressive death of segmental motoneurons (MN) leading to permanent paralysis. The chances for functional recovery after ventral root avulsion are very poor owing to the loss of avulsed neurons and the long distance that surviving neurons have to re-grow axons from the spinal cord to the corresponding targets. Following unilateral avulsion of L4, L5 and L6 spinal roots in adult rats, we performed an intraspinal transplant of mesenchymal stem cells (MSC) and surgical re-implantation of the avulsed roots. Four weeks after avulsion the survival of MN in the MSC-treated animals was significantly higher than in vehicle-injected rats (45 % vs 28 %). Re-implantation of the avulsed roots in the injured spinal cord allowed the regeneration of motor axons. By combining root re-implantation and MSC transplant the number of surviving MN at 28 days post-injury was higher (60 %) than in re-implantation alone animals (46 %). Electromyographic tests showed evidence of functional re-innervation of anterior tibialis and gastrocnemius muscles by the regenerated motor axons only in rats with the combined treatment. These results indicate that MSC are helpful in enhancing neuronal survival and increased the regenerative growth of injured axons. Surgical re-implantation and MSC grafting combined had a synergic neuroprotective effect on MN and on axonal regeneration and muscle re-innervation after spinal root avulsion.     

Differential regulation of trophic factor receptor mRNAs in spinal motoneurons after sciatic nerve transection and ventral root avulsion in the rat

After sciatic nerve lesion in the adult rat, motoneurons survive and regenerate, whereas the same lesion in the neonatal animal or an avulsion of ventral roots from the spinal cord in adults induces extensive cell death among lesioned motoneurons with limited or no axon regeneration. A number of substances with neurotrophic effects have been shown to increase survival of motoneurons in vivo and in vitro. Here we have used semiquantitative in situ hybridization histochemistry to detect the regulation in motoneurons of mRNAs for receptors to ciliary neurotrophic factor (CNTF), leukemia inhibitory factor (LIF), glial cell line-derived neurotrophic factor (GDNF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3) 1-42 days after the described three types of axon injury. After all types of injury, the mRNAs for GDNF receptors (GFRalpha-1 and c-RET) and the LIF receptor LIFR were distinctly (up to 300%) up-regulated in motoneurons. The CNTF receptor CNTFRalpha mRNA displayed only small changes, whereas the mRNA for membrane glycoprotein 130 (gp130), which is a critical receptor component for LIF and CNTF transduction, was profoundly down-regulated in motoneurons after ventral root avulsion. The BDNF full-length receptor trkB mRNA was up-regulated acutely after adult sciatic nerve lesion, whereas after ventral root avulsion trkB was down-regulated. The NT-3 receptor trkC mRNA was strongly down-regulated after ventral root avulsion. The results demonstrate that removal of peripheral nerve tissue from proximally lesioned motor axons induces profound down-regulations of mRNAs for critical components of receptors for CNTF, LIF, and NT-3 in affected motoneurons, but GDNF receptor mRNAs are up-regulated in the same situation. These results should be considered in relation to the extensive cell death among motoneurons after ventral root avulsion and should also be important for the design of therapeutical approaches in cases of motoneuron death.     

Re-established micturition reflexes show differential activation patterns after lumbosacral ventral root avulsion injury and repair in rats

Previous studies have demonstrated that an acute implantation of lesioned lumbosacral ventral roots into the rat conus medullaris (CM) results in functional reinnervation of the lower urinary tract (LUT). Although the root implantation procedure results in a return of reflexive micturition, voiding efficiency (VE) remains incompletely recovered. Here, we performed a detailed urodynamic analysis of cystometry and external urethral sphincter (EUS) electromyography (EMG) recordings to determine underlying mechanisms for the incompletely recovered VE. For this purpose, adult female rats were studied at 12 weeks after a bilateral L5–S2 ventral root avulsion injury followed by an acute surgical implantation of the avulsed L6 and S1 ventral roots into the CM (n = 6). Age-matched sham-operated rats (n = 6) were included for control purposes. Compared to sham-operated controls, rats of the implanted series showed 1) reflex bladder contractions with a significantly shortened urine expulsion phase, 2) markedly decreased phasic EUS EMG activity during micturition, and 3) a pronounced bladder–sphincter dys-coordination, as demonstrated by a significantly delayed onset of the switch from low-amplitude tonic EUS EMG activity to either phasic EUS EMG activity or a large-amplitude tonic EUS EMG activity during the urine expulsion phase. Our findings provide a mechanistic explanation for the incomplete recovery of the VE following implantation of avulsed ventral roots into the spinal cord. Our future studies will aim to increase successful axonal regeneration in attempts to augment the recovery of the LUT after cauda equina injury and repair.     

Anatomical evidence for two spinal ‘afferent–interneuron–efferent’ reflex pathways involved in micturition in the rat: a ‘pelvic nerve’ reflex pathway and a ‘sacrolumbar intersegmental’ reflex pathway

We labeled interneurons in the L1–L2 and L6–S1 spinal cord segments of the rat that are involved in bladder innervation using transneuronal retrograde transport of pseudorabies virus (PRV) in normal animals and in animals with selected nerve transections. Preganglionic neurons were identified using antisera against choline acetyltransferase (ChAT). In some experiments we labelled parasympathetic preganglionic neurons (PPNs) in the L6–S1 spinal cord by retrograde transport of Fluorogold from the major pelvic ganglion. We identified bladder afferent terminals using the transganglionic transport of the anterograde tracer cholera toxin subunit b. We present anatomical evidence for two spinal pathways involved in innervation of the bladder. First, in the intact rat, afferent information from the bladder connects, via interneurons in L6–S1, to the PPNs that provide the efferent innervation of the bladder. The afferent terminals were located mainly in close apposition to interneurons located dorsal to the retrogradely labeled PPNs. Second, using L6–S1 ganglionectomies or L6–S1 ventral root rhizotomies we limited viral transport to the sympathetic pathways innervating the bladder. This procedure also labelled interneurons (but not PPNs) with PRV in the L6–S1 spinal cord in a location very similar to those described in the intact rat. These interneurons also receive bladder afferent terminals but we propose that they project to sympathetic preganglionic neurons, most of which are in the L1–L2 spinal segments. Based on this anatomical evidence, we propose the existence of two spinal reflex pathways involved in micturition: a pathway limited to a reflex arc in the pelvic nerve (presumably excitatory to the detrusor muscle); and a pathway involving the pelvic nerve and sympathetic nerve fibers, some of which may travel in the hypogastric (presumably inhibitory to the detrusor muscle).     

Effects of C8 ventral root avulsion or transection on spinal alpha motoneurons in adult rats★○ A qualitative light and electron microscopic study

BACKGROUND： Nerve root avulsion is a frequent finding in patients with brachial plexus injury following road traffic accidents or as a result of severe arm traction during complicated deliveries. This injury constitutes a challenging clinical and surgical problem. The morphological characteristics of motoneurons after nerve root avulsion deserve further analysis.
OBJECTIVE： To study the different morphological changes of α-motoneurons under light and electron microscopy after C8 spinal ventral rootlets avulsion and transection at various stages. DESIGN： Controlled animal study.
SETTING： Department of Anatomy, King Faisal University.
MATERIALS： The experiment was carried out at the Department of Anatomy, College of Medicine, King Faisal University between January 2005 and March 2006. Six adult Sprague Dawley rats weighing 200-350 g, irrespective of gender, were used for this study. The animals were bred at the animal house, College of Medicine, King Faisal University, and fed on rat maintenance diet. Water and standard diet were supplied ad libitum. Animal interventions were carried out according to animal ethical standards.
METHODS： Three animals were randomly chosen for avulsion of the right ventral rootlets of C8 spinal nerves. The other three received transection of the right ventral rootlets of C8 spinal nerves. ①Avulsion experiment： After rats were anesthetized, the right ventral rootlets of C8 spinal nerves were identified. The ventral rootlets were avulsed from the spinal cord by traction with a fine hook （Fine Science Tools Inc., No. 10031-13, Germany）. Traction was exerted in a direction parallel to the course of the spinal root. Under the operating microscope, the C8 segment was exactly located. After checking the successfulness of the surgical procedure, the C8 segment was separated from the spinal cord. The outcome of the avulsion procedure was as follows： two animals had true avulsion, i.e., no remaining stump was attached to the spinal cord surface. One rat had a stump     

Changes in compressed neurons from dogs with acute and severe cauda equina constrictions following intrathecal injection of brain-derived neurotrophic factor-conjugated polymer nanoparticles

This study established a dog model of acute multiple cauda equina constriction by experimental constriction injury (48 hours) of the lumbosacral central processes in dorsal root ganglia neurons. The repair effect of intrathecal injection of brain-derived neurotrophic factor with 15 mg encapsulated biodegradable poly(lactide-co-glycolide) nanoparticles on this injury was then analyzed. Dorsal root ganglion cells (L7) of all experimental dogs were analyzed using hematoxylin-eosin staining and immunohistochemistry at 1, 2 and 4 weeks following model induction. Intrathecal injection of brain-derived neurotrophic factor can relieve degeneration and inflammation, and elevate the expression of brain-derived neurotrophic factor in sensory neurons of compressed dorsal root ganglion. Simultaneously, intrathecal injection of brain-derived neurotrophic factor obviously improved neurological function in the dog model of acute multiple cauda equina constriction. Results verified that sustained intraspinal delivery of brain-derived neurotrophic factor encapsulated in biodegradable nanoparticles promoted the repair of histomorphology and function of neurons within the dorsal root ganglia in dogs with acute and severe cauda equina syndrome.