Correlation between myofibrillar ATPase activity and myosin heavy chain composition in equine skeletal muscle and the influence of training.
Abstract: The histochemical myofibrillar ATPase (mATPase) method is used routinely for identification of equine skeletal muscle fiber types, but important problems have been observed with the subdivision of fast fiber population when using this method. To verify the use of this qualitative method, a number of equine muscle biopsies were analyzed with a combination of histochemical, immunohistochemical, electrophoretic, and morphometric techniques. The influence of training on these interrelations was also evaluated. Methods: Five young (2-3 years old) thoroughbred horses were intensively trained for 8 months on a high-speed treadmill. Biopsies were taken from the gluteus medius muscle at the beginning, after 4 months, and at the end of the training program. Serial sections of the samples were stained by mATPase histochemistry and immunohistochemistry by using a number of monoclonal antibodies specific to selected myosin heavy chain (MyHC) isoforms. The histochemical and immunohistochemical categorization of a large number of fibers (N = 2,078) was compared fiber by fiber. The MyHC content of homogenates of the same biopsies were quantified by densitometry of a sensitive gel electrophoretic technique and compared with histochemical and immunohistochemical fiber types. Results: A large proportion of fibers examined (approximately 20%) were misclassified by traditional mATPase histochemistry. Many fibers histochemically identified as type IIB displayed both type IIa and type IIb MyHC isoforms, and nearly all type IIAB fibers in mATPase contained only the type IIa MyHC isoform by immunohistochemistry. Correlation analyses suggested a weak relation between the histochemically assessed relative cross-sectional area occupied by the three major fiber types (I, IIA, and IIB) and the electrophoretically assessed MyHC content, whereas a stronger relation was found between immunohistochemically defined fiber types and electrophoretic data. The four fiber type populations delineated according to MyHC content (I, IIA, IIAB, and IIB) had sizes and oxidative capacities significantly different from each other. No adaptation of any parameter measured to training was found. Training had no significant effect on the number of fibers misclassified by mATPase histochemistry. Conclusions: These data demonstrate a significant limitation in mATPase histochemistry for assessing fibers containing fast MyHC isoforms. The use of monoclonal antibodies against specific MyHC isoforms seems to be a more sensitive and less subjective method.
Publication Date: 1996-10-01 PubMed ID: 8888961DOI: 10.1002/(SICI)1097-0185(199610)246:2<195::AID-AR6>3.0.CO;2-0Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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This research article explores how the standard method of identifying equine muscle fiber types using myofibrillar ATPase (mATPase) may not provide accurate results especially for classifying fast muscle fibers. The study examined muscle biopsies from thoroughbred horses under an intensive training programme and compared the mATPase method with several others including immunohistochemistry and electrophoresis. It found that around 20% of muscle fibers were misclassified using mATPase. The research concludes that using monoclonal antibodies against specific myosin heavy chain (MyHC) isoforms is a more precise method when analyzing muscle fibers.
Methodology
- The research involved five thoroughbred horses aged 2-3 years old, which underwent an intense 8-month training regimen on a high-speed treadmill.
- Muscle biopsies were taken from the gluteus medius muscle at the beginning, after 4 months, and at the end of the training.
- The samples were analysed using a combination of mATPase histochemistry, immunohistochemistry using monoclonal antibodies specific to selected MyHC isoforms, electrophoretic techniques and morphometric techniques.
Results
- Approximately 20% of the muscle fibers were misclassified by the conventional mATPase histochemical method.
- Fibers initially categorised as type IIB usually displayed type IIa and IIb MyHC isoforms and nearly all type IIAB fibers comprised only the type IIa MyHC isoform.
- Overall, the correlation between the histochemically analysed results and the densitometrically quantified MyHC content was weak.
- The correlation between the results from immunohistochemistry and electrophoresis was stronger, suggesting a more accurate classification method.
- The trainings did not produce any significant change in the muscle fibers or the frequency of misclassification by mATPase histochemical method.
Conclusions
- The limitations of the mATPase histochemical method for analyzing fibers with fast MyHC isoforms are significant.
- Applying monoclonal antibodies against specific MyHC isoforms, as is done in immunohistochemistry, offers a more sensitive and less subjective classification method for muscle fibers.
Cite This Article
APA
Rivero JL, Talmadge RJ, Edgerton VR.
(1996).
Correlation between myofibrillar ATPase activity and myosin heavy chain composition in equine skeletal muscle and the influence of training.
Anat Rec, 246(2), 195-207.
https://doi.org/10.1002/(SICI)1097-0185(199610)246:2<195::AID-AR6>3.0.CO;2-0 Publication
Researcher Affiliations
- Department of Comparative Anatomy and Pathological Anatomy, Faculty of Veterinary Science, University of Cordoba, Spain.
MeSH Terms
- Animals
- Electrophoresis, Polyacrylamide Gel
- Female
- Histocytochemistry
- Horses / anatomy & histology
- Horses / metabolism
- Immunohistochemistry
- Male
- Muscle Fibers, Skeletal / classification
- Muscle Fibers, Skeletal / metabolism
- Muscle, Skeletal / anatomy & histology
- Muscle, Skeletal / metabolism
- Myosin Heavy Chains / metabolism
- Myosins / metabolism
- Physical Conditioning, Animal
- Succinate Dehydrogenase / metabolism
Grant Funding
- N01-HD-2-3144 / NICHD NIH HHS
Citations
This article has been cited 17 times.- 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.
- Latham CM, Guy CP, Wesolowski LT, White-Springer SH. Fueling equine performance: importance of mitochondrial phenotype in equine athletes. Anim Front 2022 Jun;12(3):6-14.
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- Song S, Ahn CH, Kim GD. Muscle Fiber Typing in Bovine and Porcine Skeletal Muscles Using Immunofluorescence with Monoclonal Antibodies Specific to Myosin Heavy Chain Isoforms. Food Sci Anim Resour 2020 Jan;40(1):132-144.
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