Assessment of the rate of spinal motor axon regeneration by choline acetyltransferase immunohistochemistry following sciatic nerve crush injury in mice

Abstract

Object. The purpose of this study was to examine whether choline acetyltransferase (ChAT) staining can be used for assessing the rate of motor neuron regeneration at an early phase of axon outgrowth.Methods. The authors developed a new sciatic nerve crush model in adult mice. In this model, in addition to performing a sciatic nerve crush injury, the authors excised the ipsilateral lumbar L3-6 dorsal root ganglion (DRG), which resulted in degeneration of the sensory fibers entering into the sciatic nerve. Crushed nerve sections obtained at Day 3 or Day 7 postinjury were analyzed by means of immunostaining.Results. The immunostaining showed that ChAT, a motor axon-specific antigen, was totally co-localized with growth-associated protein 43 (GAP-43), which is expressed in regenerating nerves and transported into growth cones.Conclusions. Our results suggest that measuring the length of motor axon outgrowth by ChAT immunostaining is reliable. ChAT staining provides a more convenient method for evaluating the rate of motor axon outgrowth in a mixed nerve. (http://thejns.org/doi/abs/10.3171/2013.8.JNS121648) KeY WorDS • nerve injury • axon regeneration • motor neuron • ChAT staining • peripheral nerveAbbreviations used in this paper: CGRP = calcitonin generelated peptide; ChAT = choline acetyltransferase; GAP-43 = growth-associated protein 43; NF200 = neurofilament-200; PBS = phosphate-buffered saline.

Intravascular innate immune cells reprogrammed via intravenous nanoparticles to promote functional recovery after spinal cord injury

“…A number of genes associated with motor neurons were observed to be up-regulated at later time points. Choline acetyltranferase (ChAT), which marks mature motor neurons (43), had increased expression. Similarly, an increased expression of Homeobox D10 (Hoxd10) was observed, which organizes the patterning of motor neurons in the spinal cord (20).…”

Roles of nitric oxide and ethyl pyruvate after peripheral nerve injury

“…Acetylcholinesterase histochemistry was used to examine WD. The hindered nerve regeneration in iNOS-KO mice foreground the significance of iNOS expression and liberation of NO after injury underpin the notion that effective regeneration in PNS depend on the exact timeframe of cellular and molecular degeneration procedure [ 18 , 45 , 46 ]. NO is also found to be connected with the removal of misdirected axons, and absence of NO or knocking out NOS also causes the disturbance in the pruning mechanism [ 47 ].…”

Assessment of Nerve Regeneration through a Novel Microchannel Array

“…The mixed distribution of motor and sensory fibers by channel provides insight into the way in which nerve fibers regenerate through microchannels. Whereas in the native nerve, alpha motor neurons projecting to the same muscle tend to travel together and sensory fibers within the nerve tend to travel together, there appears to be a loss of organization within the regenerating channels [27,33]. Although there appears to be a relationship between larger axon diameters and CHAT+ fibers, there does not appear to be a relationship between larger number of fibers and type of fibers within each channel.…”

“…To assess for axon variation across individual channels by molecular subtype, double immunofluorescence of samples was conducted with anti-NF200 to stain for individual axons, anti-CHAT to stain for cholinergic motor axons [26,27], and DAPI, to assess for fiber distribution by channel post-regeneration (Figure 5a). Certain channels exhibited predispositions to CHAT+ fibers (Figure 5b), CHAT-fibers (Figure 5c), or low axon counts altogether ( Figure 5d).…”