Effects of repetitive conditioning crush lesions on regeneration of the rat sciatic nerve

doi:10.1016/0006-8993(90)90676-3

Brain Research

Volume 530, Issue 1, 15 October 1990, Pages 167-169

https://doi.org/10.1016/0006-8993(90)90676-3 Get rights and content

Abstract

The effect of repetitive conditioning lesions was tested on regeneration of the rat sciatic nerve. The nerve was conditioned by crush lesions one, two or three times with an interval of 2 or 4 days between each successive lesion. Axonal elongation was measured 3 days after a final test crush lesion. Two conditioning lesions stimulated axonal elongation more than one, while a third conditioning lesion had no further effect on axonal outgrowth. However, if the number of conditioning lesions were varied within a constant conditioning interval, outgrowth after the test lesion was the same. This suggests that the conditioning interval and not the number of conditioning lesions determined the outgrowth after a test lesion. When the conditioning lesion(s) and the test lesion were made at the same place, outgrowth was longer than if the lesions were spatially separated. Incorporation of [³H]thymidine in the regenerated nerve segmment showed that proliferation of non-neuronal cells was initiated by each lesion. By counting the number of cell nuclei this proliferation was shown to correspond to an increase of cells in the regenerating nerve. It is therefore possible that the greater number of non-neuronal cells in the distal nerve segment accounts for the enhanced conditioning lesion effect in nerves where the conditioning and test lesions are made at the same place.

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Cited by (26)

Crush injury to motor nerves in the G93A transgenic mouse model of amyotrophic lateral sclerosis promotes muscle reinnervation and survival of functionally intact nerve-muscle contacts
2018, Neurobiology of Disease
Citation Excerpt :
The conditioning lesion was first described in 1973 (McQuarrie and Grafstein, 1973). It is a nerve injury simultaneous with or preceding a nerve lesion that accelerates axon outgrowth from the nerve proximal to the lesion as well as accelerates the rate of regeneration through the denervated distal nerve stump (Forman et al., 1980; Lankford et al., 1998; McQuarrie, 1981; McQuarrie et al., 1978; Sjoberg and Kanje, 1990; Wu et al., 2007). Yet, in both our study and that in the SOD1G93A transgenic rat in which sciatic nerve crush injury promoted motoneuron survival, all the motoneurons that innervate the hindlimb muscles did not lose their nerve-muscle contacts before the so-called ‘conditioning’ crush injury.
Selective survival of small motor nerve fibers and their neuromuscular contacts in the SOD1^G93A transgenic mouse model of amyotrophic lateral sclerosis (ALS) suggests that smaller regenerated nerve fibers are more able to sustain reformed nerve-muscle connections as functionally intact motor units (MUs). The sciatic nerve was crushed unilaterally in SOD1^G93A transgenic mice at 40 days of age and contractile forces of reinnervated muscles and their MUs were recorded at 90 days in order to determine the capacities of the nerves to regenerate and to form and retain functional neuromuscular connections. Reduced MU numbers in fast-twitch tibialis anterior, extensor digitorum longus and medial gastrocnemius muscles and the lesser reductions in slow-twitch soleus muscle of SOD1^G93A transgenic mice were reversed in reinnervated muscles: there were more reinnervated MUs and their contractile forces and the muscle forces and weights increased. In line with the contrasting ability of only small not large nerve fibers to sprout to form enlarged MUs in the SOD1^G93A transgenic mouse, the smaller regenerating nerve fibers formed enlarged MUs that were better able to survive. Because nerve fibers with and without muscle contacts were severed by the sciatic nerve crush injury, the conditioning lesion is untenable as the explanation for improved maintenance of reinnervated neuromuscular junctions. Elevated neurotrophic factor expression in axotomized motoneurons and/or denervated Schwann cells and the synapse withdrawal from axotomized motoneurons are other factors that, in addition to reduced size of nerve fibers reinnervating muscles, may account for increased survival and size of reinnervated MUs in ALS.
Geldanamycin accelerated peripheral nerve regeneration in comparison to FK-506 in vivo
2012, Neuroscience
Citation Excerpt :
Downstream programs of neurite outgrowth are initiated, including reorganization of microtubules and microfilaments, upregulated expression of GAP-43 (a growth cone path-finding protein associated with nerve growth and plasticity) (Gold et al., 1998; Madsen et al., 1998), and potential crosstalk with signal transduction pathways for neurotrophic factors like NGF (Gold et al., 1999). As has been noted previously (Yan et al., 2011), the first injury in the saphenous nerve crush model induces a pre-conditioned nerve injury state that enhances the regenerative response to the second crush injury (McQuarrie, 1978; Forman et al., 1980; Sjoberg and Kanje, 1990; Lankford et al., 1998). Like FK-506, GA provides an additional benefit to nerve regeneration above that of pre-conditioning.
FK-506 accelerates nerve regeneration and improves functional recovery in vivo; its immunosuppressive properties, however, limit its clinical utility. Geldanamycin (GA), a non-immunosuppressive agent, shares a common binding target (heat shock protein 90) with FK-506 and may accelerate nerve regeneration through a similar mechanism. GA has been shown to augment neurite outgrowth in vitro but has not been tested in vivo. The current study investigated the effect of GA on the rate of axonal regeneration and functional recovery following peripheral nerve injury. In the first experiment, Thy1-GFP transgenic rats underwent serial transmuscular imaging to quantify the rate of axonal regeneration following saphenous nerve crush injury. In subsequent experiments, Lewis rats underwent tibial nerve crush or transection-and-repair injuries and were assessed for functional recovery by walking track analysis. All animals were randomized to receive daily administration of FK-506 (2 mg/kg), GA (0.2 mg/kg), or a control vehicle (dimethyl sulfoxide, 1 mL/kg) starting 3 days prior to injury. Both GA and FK-506 significantly increased the rate of axonal regeneration following crush injury in Thy1-GFP rats. In Lewis rats undergoing tibial nerve crush injury, earlier functional recovery occurred at day 5 and day 6 in animals treated with FK-506 and GA respectively, vs. day 13 for controls. Over a truncated 21-day timeframe, Lewis rats undergoing tibial nerve transection-and-repair injury and treated with FK-506 regained function at day 16, whereas those treated with GA or the control vehicle did not regain normal function. GA-treated animals, however, did exhibit significant functional improvement vs. controls. The current study demonstrated that GA accelerates axonal regeneration and enhances functional recovery in vivo. Its ability to increase the rate at which peripheral axons regenerate is comparable to that of FK-506. GA, however, did not match the performance of FK-506 in injury models where Wallerian degeneration (WD) is ongoing in the distal stump. This provides evidence that FK-506 accelerates axonal regeneration through two parallel mechanisms: the first being its well-established effect on neurons; the second is likely a newly described, as-yet poorly defined mechanism that affects WD. Finally, given the decrease in observed toxicity with GA administration, it might be a suitable non-immunosuppressive alternative to FK-506 for accelerating peripheral nerve regeneration in cases of clinical nerve injury.
Evaluation of peripheral nerve regeneration via in vivo serial transcutaneous imaging using transgenic Thy1-YFP mice
2011, Experimental Neurology
Citation Excerpt :
However, the first crush has a significant impact on the regenerative capacity of neurons from the proximal stump. It is well known that a prior – or preconditioning – injury enhances the regenerative capacity of a nerve for a subsequent injury (Forman et al., 1980; Lankford et al., 1998; McQuarrie, 1978; Sjoberg and Kanje, 1990). Preconditioning is associated with increased expression levels of ERK1/2 and GAP43 (Perlson et al., 2005).
This study uses the saphenous nerve crush model in Thy1-YFP mice and serial transcutaneous imaging to evaluate the rate of nerve regeneration under various FK-506 (tacrolimus) dosing regimens and in the presence of transgenic overexpression of glial cell line-derived neurotrophic factor (GDNF). Thy1-YFP transgenic mice received saphenous nerve crush and were monitored for axonal regeneration via transcutaneous imaging for 7 days. Group A received no FK-506. Groups B and C received FK-506 at 2 or 0.5 mg/kg/day, starting three days before injury (preload). Groups D and E received FK-506 at 2 or 0.5 mg/kg/day, starting on the day of injury. Group F consisted of double transgenic mice with central overexpression of GDNF by CNS astrocytes (GFAP-GDNF/Thy1-YFP). Length and rate of axonal regeneration were measured and calculated over time. Regardless of concentration, FK-506 preload (Groups B and C) improved length and rate of axonal outgrowth compared with controls (Group A) and no preload (Groups D and E). Surprisingly, central overexpression of GDNF (GFAP-GDNF) delayed and stunted axonal outgrowth. Saphenous nerve crush in Thy1-YFP mice represents a viable model for timely evaluation of therapeutic strategies affecting the rate of nerve regeneration. FK-506 administered three days prior to injury accelerates axonal regeneration beyond injury conditioned regeneration alone and may serve as a reliable positive control for the model. GDNF overexpression in the CNS impedes early axonal outgrowth.
A conditioning lesion enhances sympathetic neurite outgrowth
2005, Experimental Neurology
Axonal regeneration can be influenced by a conditioning lesion (an axonal injury made prior to a second test lesion). Previously, sympathetic neurons in vivo were shown to respond to a conditioning lesion with decreased neurite outgrowth, in contrast to the enhanced outgrowth observed in all other peripheral neurons examined. The present experiments tested the effects of a conditioning lesion on neurite outgrowth in vitro from the superior cervical ganglion (SCG) and the impact of several factors on that response. Ganglia axotomized 1 week earlier and then explanted in Matrigel or collagen gel responded with a significant increase in neurite extension compared to sham-operated ganglia. A distal axotomy produced by unilateral removal of the salivary glands (sialectomy) caused an increase in neurite outgrowth similar to that of a proximal axotomy. These conditioning lesions induced both an increase in the rate of elongation, and, in the case of the proximally axotomized SCG, a shorter initial delay of outgrowth. The enhanced outgrowth following sialectomy was specific to the nerve containing the majority of axons projecting to the salivary glands, suggesting that the conditioning lesion effect is restricted to previously injured neurons. Deletion of the gene for leukemia inhibitory factor (LIF), a gene induced by axotomy, did not abolish the conditioning lesion effect in SCG explants or dissociated cell cultures. In conclusion, sympathetic neurons are capable of responding to a conditioning lesion with increased neurite outgrowth. The hypothesis that the neuronal cell body response to axotomy plays an important role in the conditioning lesion response is discussed.
Involvement of α7β1 integrin in the conditioning-lesion effect on sensory axon regeneration
2003, Molecular and Cellular Neuroscience
Conditioning lesions of peripheral nerves improve axonal regeneration after injury and involve changes in expression of proteins required for axonal growth. Integrin α7β1 expression in motor and sensory neurons increases following nerve lesions and motor axon regeneration is impaired in α7 integrin KO mice (J. Neurosci. 20, 1822–1830). To investigate the role of α7β1 integrin in sensory axon regeneration, dorsal root ganglia of adult mice were cultured in gels of laminin-rich extracellular matrix (Matrigel) or collagen. Normal dorsal root ganglia in Matrigel or collagen supplemented with laminin showed spontaneous axonal outgrowth, which was greatly increased in conditioned preparations, but only in the presence of laminin. Conditioned dorsal root ganglia from normal mice cultured with a blocking antibody to β1 integrin and from α7 integrin KO mice showed reduced axonal growth in both Matrigel- and laminin-supplemented collagen gels. Enhanced axonal regeneration after conditioning lesions therefore involves increased responsiveness to laminin and integrin α7β1 expression.
Respective roles of inflammation and axonal breakdown in the regulation of peripheral nerve hemopexin: An analysis in rats and in C57BL/Wlds mice
2000, Journal of Neuroimmunology
We have previously demonstrated that one of the peripheral nerve responses to injury is the overexpression of hemopexin (HPX). Here, we demonstrate that Wallerian degeneration is required for this response, since HPX does not increase in C57BL/Wlds mice, which display a severely impaired Wallerian degeneration. We also show that HPX synthesis is dramatically increased in macrophages during their activation or after IL-6 stimulation. However, IL-6-driven HPX overexpression occurs in vivo and in vitro in the absence of substantial macrophage invasion. We conclude that, after nerve injury, HPX overexpression occurs first in Schwann cells as a result of axotomy and is subsequently regulated by inflammation. Furthermore, our results and those already described suggest that IL-6, synthesized by the various cell types producing HPX, control nerve HPX expression via paracrine and autocrine mechanisms.

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: This study was supported by grants from the Swedish Natural Science Research Council and Hierta-Retzius Stiftelse.

¹: We wish to thank Marie Adlher-Maihofer for her skillful technical assistance and Marianne Andersson for excellent artwork.

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Effects of repetitive conditioning crush lesions on regeneration of the rat sciatic nerve

Abstract

Brain Research

Brain Research

Exp. Neurol.

Brain Research

Brain Research

Brain Research

Brain Research

Brain Research

Brain Research

An analysis of growth in nuclear population during Wallerian degeneration

J. Cell. Comp. Physiol.

Regeneration of the rat sciatic nerve after different conditioning lesions: effects of the conditioning interval

Microsurgery

Increased regeneration rate in peripheral nerve axons following double lesions: enhancement of the conditioning lesion phenomenon

J. Neurobiol.