Multiple immunofluorescence labelling enables simultaneous identification of all mature fibre types in a single equine skeletal muscle cryosection.
Abstract: Skeletal muscle is composed of a heterogeneous mixture of several fibre types, each with specific physiological properties. In equine muscle, identification of these individual fibres (fibre typing) is important for both exercise physiology and pathological studies. Traditionally, fibre typing has been achieved by adenosine triphosphatase (ATPase) histochemistry or by immunoperoxidase labelling with antibodies directed at myosin heavy chain isoforms. ATPase histochemistry can be temperamental and lacks specificity, and both techniques require staining of serial cryosections to reveal the entire fibre type compliment of a single sample, which is time consuming and prone to inaccuracy. Here we describe an immunofluorescence labelling technique that enables rapid, accurate and specific identification of the 3 mature equine muscle fibre types in a single cryosection.
© 2011 EVJ Ltd.
Publication Date: 2011-03-04 PubMed ID: 21496090DOI: 10.1111/j.2042-3306.2010.00329.xGoogle Scholar: Lookup
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Summary
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The research study introduces a novel immunofluorescence labelling method that allows for swift and precise identification of different types of mature muscle fibres in a single sample from equine skeletal muscle.
Overview of the Study
- The article focuses primarily on skeletal muscle, which is a heterogeneous combination of various fibre types, each possessing unique physiological properties. In the context of equine muscle, identifying these individual fibres, a process known as fibre typing, bears significant importance for both exercise physiology and pathological investigations.
- In the past, fibre typing was typically accomplished using adenosine triphosphatase (ATPase) histochemistry or immunoperoxidase labelling with antibodies that target myosin heavy chain isoforms.
Limits of Traditional Methods
- The research articulates how traditional methods like ATPase histochemistry can be unpredictable and lack specificity. Both ATPase histochemistry and immunoperoxidase labelling necessitate staining of sequential cryosections to expose the full range of fibre types in a single sample.
- This serial staining process is both time-consuming and prone to inaccuracies, proving to be less efficient.
New Immunofluorescence Labelling Technique
- The study introduces a new method using immunofluorescence labelling, which overcomes the limitations of traditional methods.
- This innovative technique offers rapid, accurate, and specific identification of the three mature equine muscle fibre types within a single cryosection.
- Through this method, researchers may eliminate the need for exhaustive staining of sequential cryosections, saving both time and minimizing the risk of inaccuracies.
Implications of the Study
- The utilization of this enhanced methodology could significantly impact exercise physiology and pathological studies relating to equine muscle.
- By allowing for a more efficient and precise identification of muscle fibre types, it can contribute to improved understanding of muscle composition, functionality, and any potential pathological deviations.
Cite This Article
APA
Tulloch LK, Perkins JD, Piercy RJ.
(2011).
Multiple immunofluorescence labelling enables simultaneous identification of all mature fibre types in a single equine skeletal muscle cryosection.
Equine Vet J, 43(4), 500-503.
https://doi.org/10.1111/j.2042-3306.2010.00329.x Publication
Researcher Affiliations
- Comparative Neuromuscular Diseases Laboratory, Department of Veterinary Clinical Sciences, Royal Veterinary College, London, UK.
MeSH Terms
- Animals
- Antibodies, Monoclonal / chemistry
- Fluorescent Antibody Technique / methods
- Fluorescent Antibody Technique / veterinary
- Horses / anatomy & histology
- Muscle Fibers, Skeletal / cytology
Citations
This article has been cited 13 times.- Cahalan SD, Perkins JD, Boehm I, Jones RA, Gillingwater TH, Piercy RJ. A method to identify, dissect and stain equine neuromuscular junctions for morphological analysis. J Anat 2022 Nov;241(5):1133-1147.
- Busse NI, Gonzalez ML, Wagner AL, Johnson SE. Short Communication: Supplementation with calcium butyrate causes an increase in the percentage of oxidative fibers in equine gluteus medius muscle. J Anim Sci 2022 Aug 1;100(8).
- Latham CM, Owen RN, Dickson EC, Guy CP, White-Springer SH. Skeletal Muscle Adaptations to Exercise Training in Young and Aged Horses. Front Aging 2021;2:708918.
- Busse NI, Gonzalez ML, Krason ML, Johnson SE. β-Hydroxy β-methylbutyrate supplementation to adult Thoroughbred geldings increases type IIA fiber content in the gluteus medius. J Anim Sci 2021 Oct 1;99(10).
- Williams ZJ, Velez-Irizarry D, Gardner K, Valberg SJ. Integrated proteomic and transcriptomic profiling identifies aberrant gene and protein expression in the sarcomere, mitochondrial complex I, and the extracellular matrix in Warmblood horses with myofibrillar myopathy. BMC Genomics 2021 Jun 11;22(1):438.
- Valberg SJ, Henry ML, Perumbakkam S, Gardner KL, Finno CJ. An E321G MYH1 mutation is strongly associated with nonexertional rhabdomyolysis in Quarter Horses. J Vet Intern Med 2018 Sep;32(5):1718-1725.
- Carraro U. Exciting perspectives for Translational Myology in the Abstracts of the 2018Spring PaduaMuscleDays: Giovanni Salviati Memorial - Chapter III - Abstracts of March 16, 2018. Eur J Transl Myol 2018 Jan 12;28(1):7365.
- Finno CJ, Gianino G, Perumbakkam S, Williams ZJ, Bordbari MH, Gardner KL, Burns E, Peng S, Durward-Akhurst SA, Valberg SJ. A missense mutation in MYH1 is associated with susceptibility to immune-mediated myositis in Quarter Horses. Skelet Muscle 2018 Mar 6;8(1):7.
- Wu P, Zhang S, Spinner RJ, Lizardi MT, Gu Y, Yu C, Yaszemski MJ, Windebank AJ, Wang H. A novel triple immunoenzyme staining enables simultaneous identification of all muscle fiber types on a single skeletal muscle cryosection from normal, denervated or reinnervated rats. Neural Regen Res 2017 Aug;12(8):1357-1364.
- Karbiener M, Jarvis JC, Perkins JD, Lanmüller H, Schmoll M, Rode HS, Gerstenberger C, Gugatschka M. Reversing Age Related Changes of the Laryngeal Muscles by Chronic Electrostimulation of the Recurrent Laryngeal Nerve. PLoS One 2016;11(11):e0167367.
- Valberg SJ, Williams ZJ, Ames EG, Mickelson JR, Nout-Lomas YS, Landolt G, Sanz M, Gardner K. Aberrant skeletal muscle morphogenesis and myofiber differentiation characterize equine myotonic dystrophy. PLoS One 2026;21(1):e0341655.
- Kirsch A, Gerstenberger C, Jakubaß B, Tschernitz M, Perkins JD, Groselj-Strele A, Lanmüller H, Jarvis JC, Kniesburges S, Döllinger M, Gugatschka M. Bilateral Functional Electrical Stimulation for the Treatment of Presbyphonia in a Sheep Model. Laryngoscope 2024 Feb;134(2):848-854.
- Lean NE, Franklin SH, Steel C, Woolford L, White J, Ahern BJ. Evaluation of recurrent laryngeal neuropathy in domestic and feral horse populations in Australia using histologic and immunohistochemical analysis: A pilot study. Vet Med Sci 2023 Jul;9(4):1610-1617.
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