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Analysis of myosin heavy chains at the protein level in horse skeletal muscle.
J Muscle Res Cell Motil. 1999 Feb; 20(2):211-21.JM

Abstract

Combined methodologies of enzyme-linked immunosorbent assay (ELISA), sodium dodecyl sulphate polyacrilamide gel electrophoresis (SDS-PAGE), immunoblotting, traditional myofibrillar ATPase (mATPase) histochemistry and immunocytochemistry of whole biopsied samples were used to study myosin heavy chain (MHC) isoforms in the equine gluteus medius muscle. The ELISA technique allowed the quantification of the three MHC isoforms known to be present in different horse muscles: slow (MHC-I) and two fast (termed MHC-IIA and MCH-IIX). The SDS-PAGE method resolved MHCs in three bands: MHC-I, MHC-IIX and MHC-IIA from the fastest to the slowest migrating band and a quantification by densitometry for each MHC isoform was also possible. The identity of these three MHCs was confirmed by immunoblots with specific monoclonal antibodies. Five fibre types were defined immunohistochemically according to their MHC content: I, I + IIA, IIA, the hybrid IIAX and IIX. When quantitative data obtained with the four different methodologies were combined and compared, they were consistent and, when considered together, showed significant correlation. Nevertheless, the percentage of MHC-IIA histochemically derived was underestimated, while that of MHC-IIX was overestimated in comparison with the immunocytochemical determination of these MHC isoforms. The percentage of MHC-I obtained by ELISA technique was underestimated. In short, these integrated methods for the analysis of MHCs at the protein level demonstrate that equine skeletal muscle does not express the MHC-IIB, so type II fibres have been misclassified in numerous previous studies based upon the vary traditional mATPase histochemistry. They also offer new prospects for muscle fibre typing in equine experimental studies and veterinary medicine.

Authors+Show Affiliations

Department of Anatomy, Faculty of Veterinary Science, University of Cordoba, Spain. anllorij@lucano.uco.esNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

10412092

Citation

Rivero, J L., et al. "Analysis of Myosin Heavy Chains at the Protein Level in Horse Skeletal Muscle." Journal of Muscle Research and Cell Motility, vol. 20, no. 2, 1999, pp. 211-21.
Rivero JL, Serrano AL, Barrey E, et al. Analysis of myosin heavy chains at the protein level in horse skeletal muscle. J Muscle Res Cell Motil. 1999;20(2):211-21.
Rivero, J. L., Serrano, A. L., Barrey, E., Valette, J. P., & Jouglin, M. (1999). Analysis of myosin heavy chains at the protein level in horse skeletal muscle. Journal of Muscle Research and Cell Motility, 20(2), 211-21.
Rivero JL, et al. Analysis of Myosin Heavy Chains at the Protein Level in Horse Skeletal Muscle. J Muscle Res Cell Motil. 1999;20(2):211-21. PubMed PMID: 10412092.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Analysis of myosin heavy chains at the protein level in horse skeletal muscle. AU - Rivero,J L, AU - Serrano,A L, AU - Barrey,E, AU - Valette,J P, AU - Jouglin,M, PY - 1999/7/21/pubmed PY - 1999/7/21/medline PY - 1999/7/21/entrez SP - 211 EP - 21 JF - Journal of muscle research and cell motility JO - J Muscle Res Cell Motil VL - 20 IS - 2 N2 - Combined methodologies of enzyme-linked immunosorbent assay (ELISA), sodium dodecyl sulphate polyacrilamide gel electrophoresis (SDS-PAGE), immunoblotting, traditional myofibrillar ATPase (mATPase) histochemistry and immunocytochemistry of whole biopsied samples were used to study myosin heavy chain (MHC) isoforms in the equine gluteus medius muscle. The ELISA technique allowed the quantification of the three MHC isoforms known to be present in different horse muscles: slow (MHC-I) and two fast (termed MHC-IIA and MCH-IIX). The SDS-PAGE method resolved MHCs in three bands: MHC-I, MHC-IIX and MHC-IIA from the fastest to the slowest migrating band and a quantification by densitometry for each MHC isoform was also possible. The identity of these three MHCs was confirmed by immunoblots with specific monoclonal antibodies. Five fibre types were defined immunohistochemically according to their MHC content: I, I + IIA, IIA, the hybrid IIAX and IIX. When quantitative data obtained with the four different methodologies were combined and compared, they were consistent and, when considered together, showed significant correlation. Nevertheless, the percentage of MHC-IIA histochemically derived was underestimated, while that of MHC-IIX was overestimated in comparison with the immunocytochemical determination of these MHC isoforms. The percentage of MHC-I obtained by ELISA technique was underestimated. In short, these integrated methods for the analysis of MHCs at the protein level demonstrate that equine skeletal muscle does not express the MHC-IIB, so type II fibres have been misclassified in numerous previous studies based upon the vary traditional mATPase histochemistry. They also offer new prospects for muscle fibre typing in equine experimental studies and veterinary medicine. SN - 0142-4319 UR - https://www.unboundmedicine.com/medline/citation/10412092/Analysis_of_myosin_heavy_chains_at_the_protein_level_in_horse_skeletal_muscle_ L2 - http://ovidsp.ovid.com/ovidweb.cgi?T=JS&PAGE=linkout&SEARCH=10412092.ui DB - PRIME DP - Unbound Medicine ER -