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A sensitive electrophoretic method for the quantification of myosin heavy chain isoforms in horse skeletal muscle: histochemical and immunocytochemical verifications.
Electrophoresis. 1997 Oct; 18(11):1967-72.E

Abstract

In adult horses, three myosin heavy chain (MyHC) isoforms can be identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunohistochemistry using specific anti-MyHC monoclonal antibodies. This report studies the suitability of a consistent SDS-PAGE technique for quantifying MyHC profiles in homogenized cryostate sections of equine gluteus medius muscle biopsies (n = 18). The method used (previously described by R. J. Talmadge and R. R. Roy; J. Appl. Physiol. 1993, 75, 2337-2340) resolved MyHCs in three bands: I, IIB or IIX, and IIA from the fastest to the slowest migration band. The success rate of the protocol for yielding three well-differentiated MyHC bands was 100% and a subsequent quantification by densitometry for each MyHC isoform was obtained in all 18 muscle biopsies. The results obtained with this electrophoretic method were compared with routine myofibrillar adenosine triphosphatase histochemistry and immunohistochemistry using specific anti-MyHC monoclonal antibodies. The percent composition of the three electrophoretically separated MyHC isoforms (I, IIA and IIB or IIX) showed strong positive correlation with percentages of the area occupied in the biopsies by the three major fiber types (I, IIA, and IIB) identified histochemically (r = 0.96, P < 0.001) and immunohistochemically (r = 0.94, P < 0.01). It can be concluded that the electrophoretic method used here for measuring MyHC content is a valid alternative for muscle fiber typing in horses. As it is less costly and time-consuming than both qualitative histochemistry and immunohistochemistry, the method offers new prospects for application in equine experimental studies and veterinary medicine.

Authors+Show Affiliations

Department of Comparative Anatomy and Pathological Anatomy, Faculty of Veterinary Science, University of Cordoba, Spain. an1lorij@lucano.uco.esNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

9420154

Citation

Rivero, J L., et al. "A Sensitive Electrophoretic Method for the Quantification of Myosin Heavy Chain Isoforms in Horse Skeletal Muscle: Histochemical and Immunocytochemical Verifications." Electrophoresis, vol. 18, no. 11, 1997, pp. 1967-72.
Rivero JL, Talmadge RJ, Edgerton VR. A sensitive electrophoretic method for the quantification of myosin heavy chain isoforms in horse skeletal muscle: histochemical and immunocytochemical verifications. Electrophoresis. 1997;18(11):1967-72.
Rivero, J. L., Talmadge, R. J., & Edgerton, V. R. (1997). A sensitive electrophoretic method for the quantification of myosin heavy chain isoforms in horse skeletal muscle: histochemical and immunocytochemical verifications. Electrophoresis, 18(11), 1967-72.
Rivero JL, Talmadge RJ, Edgerton VR. A Sensitive Electrophoretic Method for the Quantification of Myosin Heavy Chain Isoforms in Horse Skeletal Muscle: Histochemical and Immunocytochemical Verifications. Electrophoresis. 1997;18(11):1967-72. PubMed PMID: 9420154.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A sensitive electrophoretic method for the quantification of myosin heavy chain isoforms in horse skeletal muscle: histochemical and immunocytochemical verifications. AU - Rivero,J L, AU - Talmadge,R J, AU - Edgerton,V R, PY - 1998/1/7/pubmed PY - 1998/1/7/medline PY - 1998/1/7/entrez KW - Non-programmatic SP - 1967 EP - 72 JF - Electrophoresis JO - Electrophoresis VL - 18 IS - 11 N2 - In adult horses, three myosin heavy chain (MyHC) isoforms can be identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunohistochemistry using specific anti-MyHC monoclonal antibodies. This report studies the suitability of a consistent SDS-PAGE technique for quantifying MyHC profiles in homogenized cryostate sections of equine gluteus medius muscle biopsies (n = 18). The method used (previously described by R. J. Talmadge and R. R. Roy; J. Appl. Physiol. 1993, 75, 2337-2340) resolved MyHCs in three bands: I, IIB or IIX, and IIA from the fastest to the slowest migration band. The success rate of the protocol for yielding three well-differentiated MyHC bands was 100% and a subsequent quantification by densitometry for each MyHC isoform was obtained in all 18 muscle biopsies. The results obtained with this electrophoretic method were compared with routine myofibrillar adenosine triphosphatase histochemistry and immunohistochemistry using specific anti-MyHC monoclonal antibodies. The percent composition of the three electrophoretically separated MyHC isoforms (I, IIA and IIB or IIX) showed strong positive correlation with percentages of the area occupied in the biopsies by the three major fiber types (I, IIA, and IIB) identified histochemically (r = 0.96, P < 0.001) and immunohistochemically (r = 0.94, P < 0.01). It can be concluded that the electrophoretic method used here for measuring MyHC content is a valid alternative for muscle fiber typing in horses. As it is less costly and time-consuming than both qualitative histochemistry and immunohistochemistry, the method offers new prospects for application in equine experimental studies and veterinary medicine. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/9420154/A_sensitive_electrophoretic_method_for_the_quantification_of_myosin_heavy_chain_isoforms_in_horse_skeletal_muscle:_histochemical_and_immunocytochemical_verifications_ L2 - https://doi.org/10.1002/elps.1150181115 DB - PRIME DP - Unbound Medicine ER -