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Home / Equine Research Bank / J Anat / A method to identify, dissect and stain equine neuromuscular...
Journal of anatomy2022; 241(5); 1133-1147; doi: 10.1111/joa.13747

A method to identify, dissect and stain equine neuromuscular junctions for morphological analysis.

Abstract: Morphological study of the neuromuscular junction (NMJ), a specialised peripheral synapse formed between a lower motor neuron and skeletal muscle fibre, has significantly contributed to the understanding of synaptic biology and neuromuscular disease pathogenesis. Rodent NMJs are readily accessible, and research into conditions such as amyotrophic lateral sclerosis (ALS), Charcot-Marie-Tooth disease (CMT), and spinal muscular atrophy (SMA) has relied heavily on experimental work in these small mammals. However, given that nerve length dependency is an important feature of many peripheral neuropathies, these rodent models have clear shortcomings; large animal models might be preferable, but their size presents novel anatomical challenges. Overcoming these constraints to study the NMJ morphology of large mammalian distal limb muscles is of prime importance to increase cross-species translational neuromuscular research potential, particularly in the study of long motor units. In the past, NMJ phenotype analysis of large muscle bodies within the equine distal pelvic limb, such as the tibialis cranialis, or within muscles of high fibrous content, such as the soleus, has posed a distinct experimental hurdle. We optimised a technique for NMJ location and dissection from equine pelvic limb muscles. Using a quantification method validated in smaller species, we demonstrate their morphology and show that equine NMJs can be reliably dissected, stained and analysed. We reveal that the NMJs within the equine soleus have distinctly different morphologies when compared to the extensor digitorum longus and tibialis cranialis muscles. Overall, we demonstrate that equine distal pelvic limb muscles can be regionally dissected, with samples whole-mounted and their innervation patterns visualised. These methods will allow the localisation and analysis of neuromuscular junctions within the muscle bodies of large mammals to identify neuroanatomical and neuropathological features.
© 2022 The Authors. Journal of Anatomy published by John Wiley & Sons Ltd on behalf of Anatomical Society.
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Publication Date: 2022-09-10 PubMed ID: 36087283PubMed Central: PMC9558161DOI: 10.1111/joa.13747Google Scholar: Lookup
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  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research focused on developing a method to identify, dissect, and stain equine neuromuscular junctions for morphological analysis. The study is designed to allow better understanding of synaptic biology and neuromuscular disease pathogenesis, particularly in large mammals like horses.

Background

  • The neuromuscular junction (NMJ) is a specialized peripheral synapse formed between a lower motor neuron and skeletal muscle fibre.
  • Studying its morphology provides significant insights into synaptic biology and the pathogenesis of neuromuscular diseases.
  • Previous research has often centered around smaller mammals, such as rodents, due to the accessibility of their NMJs. Diseases studied using these models include amyotrophic lateral sclerosis (ALS), Charcot-Marie-Tooth disease (CMT), and spinal muscular atrophy (SMA).
  • However, many peripheral neuropathies depend on nerve length, and therefore, these rodent models have limitations. Large animals might present a better model, but their size introduces new anatomical challenges.

The Research Process

  • To increase cross-species translational neuromuscular research potential, the researchers focused on overcoming these challenges and studying the NMJ morphology of large mammalian distal limb muscles.
  • Previously, analyzing the NMJ phenotype within large muscle bodies from equine distal pelvic limbs, such as the tibialis cranialis, or within high fibrous count muscles, like the soleus, has proved difficult.
  • The researchers were able to develop a technique for locating and dissecting NMJs from equine pelvic limb muscles.
  • Using a quantification method previously validated in smaller species, the researchers demonstrated the morphology of these junctions, showing that they can be reliably dissected, stained, and analyzed.

Results and Conclusion

  • The researchers found that NMJs within the equine soleus have distinctly different morphologies compared to those in the extensor digitorum longus and tibialis cranialis muscles.
  • In conclusion, equine distal pelvic limb muscles can be regionally dissected, with samples whole-mounted and their innervation patterns visualised. This will allow researchers to localize and analyze NMJs within the muscle bodies of large mammals.
  • This research is expected to aid in identifying neuroanatomical and neuropathological features.

Cite This Article

APA
Cahalan SD, Perkins JD, Boehm I, Jones RA, Gillingwater TH, Piercy RJ. (2022). A method to identify, dissect and stain equine neuromuscular junctions for morphological analysis. J Anat, 241(5), 1133-1147. https://doi.org/10.1111/joa.13747

Publication

Journal of anatomy
ISSN: 1469-7580
NlmUniqueID: 0137162
Country: England
Language: English
Volume: 241
Issue: 5
Pages: 1133-1147

Researcher Affiliations

Cahalan, Stephen D
  • Comparative Neuromuscular Diseases Laboratory, Department of Clinical Science and Services, Royal Veterinary College, London, UK.
Perkins, Justin D
  • Comparative Neuromuscular Diseases Laboratory, Department of Clinical Science and Services, Royal Veterinary College, London, UK.
Boehm, Ines
  • Edinburgh Medical School: Biomedical Sciences, University of Edinburgh, Edinburgh, UK.
  • Euan MacDonald Centre for Motor Neuron Disease Research, University of Edinburgh, Edinburgh, UK.
  • Biozentrum, University of Basel, Basel, Switzerland.
Jones, Ross A
  • Edinburgh Medical School: Biomedical Sciences, University of Edinburgh, Edinburgh, UK.
  • Euan MacDonald Centre for Motor Neuron Disease Research, University of Edinburgh, Edinburgh, UK.
Gillingwater, Thomas H
  • Edinburgh Medical School: Biomedical Sciences, University of Edinburgh, Edinburgh, UK.
  • Euan MacDonald Centre for Motor Neuron Disease Research, University of Edinburgh, Edinburgh, UK.
Piercy, Richard J
  • Comparative Neuromuscular Diseases Laboratory, Department of Clinical Science and Services, Royal Veterinary College, London, UK.

MeSH Terms

  • Animals
  • Coloring Agents
  • Horses
  • Mammals
  • Motor Neurons / pathology
  • Muscle Fibers, Skeletal
  • Muscle, Skeletal / pathology
  • Neuromuscular Junction / pathology
  • Peripheral Nervous System Diseases / pathology

Grant Funding

  • vet/prj/780 / Horserace Betting Levy Board

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Citations

This article has been cited 2 times.
  1. Cahalan SD, Boehm I, Jones RA, Piercy RJ. Recognising the potential of large animals for modelling neuromuscular junction physiology and disease. J Anat 2022 Nov;241(5):1120-1132.
    doi: 10.1111/joa.13749pubmed: 36056593google scholar: lookup
  2. Sleigh JN. Editorial: Peripheral nerve anatomy in health and disease. J Anat 2022 Nov;241(5):1083-1088.
    doi: 10.1111/joa.13746pubmed: 36226698google scholar: lookup
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