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

Laboratory investigation

Qiuju Yuan Ph.D. 1 , Huanxing Su Ph.D. 5 , Kin Chiu Ph.D. 2 , Zhi-Xiu Lin Ph.D. 1 , and Wutian Wu M.D., Ph.D. 2 , 3 , 4 , 6
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  • 1 School of Chinese Medicine, Faculty of Science, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong SAR, China;
  • 2 Department of Anatomy,
  • 3 State Key Laboratory of Brain and Cognitive Sciences,
  • 4 Research Center of Reproduction, Development and Growth, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China;
  • 5 State Key Laboratory of Quality Research in Chinese Medicine and Institute of Chinese Medical Sciences, University of Macau, Macao SAR, China; and
  • 6 GHM Institute of CNS regeneration, Jinan University, Guangzhou, China
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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.

Abbreviations used in this paper:CGRP = calcitonin gene–related peptide; ChAT = choline acetyltransferase; GAP-43 = growth-associated protein 43; NF200 = neurofilament-200; PBS = phosphate-buffered saline.

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Contributor Notes

Address correspondence to: Wutian Wu, M.D., Ph.D., Department of Anatomy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, 21 Sassoon Rd., Hong Kong SAR, China. email: wtwu@hkucc.hku.hk.

Please include this information when citing this paper: published online September 13, 2013; DOI: 10.3171/2013.8.JNS121648.

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