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An improved method for the simultaneous demonstration of mRNA and esterase activity at the human neuromuscular junction

Lookup NU author(s): Professor Susan Lindsay, Dr Ruth Vater, Emeritus Professor Clarke Slater

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Abstract

The aim of this study was to develop a simple means of studying the distribution of mRNA coding for post-synaptic proteins at the human neuromuscular junction. A reliable method by which to identify the junctions in tissue sections after in situ hybridization was essential. A method is described for combining tile histochemical demonstration of esterase activity at the neuromuscular junction with autoradiographic localization of mRNA by in situ hybridization in the same cryostat section of skeletal muscle. The indigogenic esterase method of Strum and Hall-Craggs (1982) was modified in such a way that it is able to survive the multiple steps involved in in situ hybridization and autoradiography. The protocol is simple and reproducible and has been used successfully on sections of both rat and human skeletal muscle. To demonstrate the method, sections were reacted to reveal esterase activity and were then processed for in situ hybridization using a 35S- labelled probe specific for the ε-subunit of the acetylcholine receptor. The reaction product was retained after the lengthy in situ hybridization and autoradiographic procedures. To our knowledge, this is the first demonstration of acetylcholine receptor mRNA by in situ hybridization at human neuromuscular junctions.


Publication metadata

Author(s): Young C, Lindsay S, Vater R, Slater CR

Publication type: Article

Publication status: Published

Journal: Histochemical Journal

Year: 1998

Volume: 30

Issue: 1

Pages: 7-11

Print publication date: 01/01/1998

ISSN (print): 0018-2214

ISSN (electronic): 1567-2387

Publisher: Springer

URL: http://dx.doi.org/10.1023/A:1003206327367

DOI: 10.1023/A:1003206327367

PubMed id: 9539201


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