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Immunolabelling, histochemistry and in situ hybridisation in human skeletal muscle fibres to detect myosin heavy chain expression at the protein and mRNA level.
J Anat. 2001 Sep; 199(Pt 3):329-37.JA

Abstract

The distribution of muscle fibres classified on the basis of their content of different myosin heavy chain (MHC) isoforms was analysed in vastus lateralis muscle biopsies of 15 young men (with an average age of 22 y) by correlating immunohistochemistry with specific anti-MHC monoclonal antibodies, myofibrillar ATPase (mATPase) histochemistry and in situ hybridisation with probes specific for MHC beta-slow, MHC-IIA and MHC-IIX. The characterisation of a large number of individual fibres was compared and correlated on a fibre-to-fibre basis. The panel of monoclonal antibodies used in the study allowed classification of human skeletal muscle fibres into 5 categories according to the MHC isoform they express at the protein level, types I, I+IIA, IIA, IIAX and IIX. Hybrid fibres coexpressing two isoforms represented a considerable proportion of the fibre composition (about 14%) and were clearly underestimated by mATPase histochemistry. For a very high percentage of fibres there was a precise correspondence between the MHC protein isoforms and mRNA transcripts. The integrated methods used demonstrate a high degree of precision of the immunohistochemical procedure used for the identification and quantification of human skeletal muscle fibre types. The monoclonal antibody S5-8H2 is particularly useful for identifying hybrid IIAX fibres. This protocol offers new prospects for muscle fibre classification in human experimental studies.

Authors+Show Affiliations

Department of Comparative Anatomy and Pathological Anatomy, University of Cordoba, Spain.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

11554510

Citation

Serrano, A L., et al. "Immunolabelling, Histochemistry and in Situ Hybridisation in Human Skeletal Muscle Fibres to Detect Myosin Heavy Chain Expression at the Protein and mRNA Level." Journal of Anatomy, vol. 199, no. Pt 3, 2001, pp. 329-37.
Serrano AL, Pérez M, Lucía A, et al. Immunolabelling, histochemistry and in situ hybridisation in human skeletal muscle fibres to detect myosin heavy chain expression at the protein and mRNA level. J Anat. 2001;199(Pt 3):329-37.
Serrano, A. L., Pérez, M., Lucía, A., Chicharro, J. L., Quiroz-Rothe, E., & Rivero, J. L. (2001). Immunolabelling, histochemistry and in situ hybridisation in human skeletal muscle fibres to detect myosin heavy chain expression at the protein and mRNA level. Journal of Anatomy, 199(Pt 3), 329-37.
Serrano AL, et al. Immunolabelling, Histochemistry and in Situ Hybridisation in Human Skeletal Muscle Fibres to Detect Myosin Heavy Chain Expression at the Protein and mRNA Level. J Anat. 2001;199(Pt 3):329-37. PubMed PMID: 11554510.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Immunolabelling, histochemistry and in situ hybridisation in human skeletal muscle fibres to detect myosin heavy chain expression at the protein and mRNA level. AU - Serrano,A L, AU - Pérez,M, AU - Lucía,A, AU - Chicharro,J L, AU - Quiroz-Rothe,E, AU - Rivero,J L, PY - 2001/9/14/pubmed PY - 2001/9/28/medline PY - 2001/9/14/entrez SP - 329 EP - 37 JF - Journal of anatomy JO - J Anat VL - 199 IS - Pt 3 N2 - The distribution of muscle fibres classified on the basis of their content of different myosin heavy chain (MHC) isoforms was analysed in vastus lateralis muscle biopsies of 15 young men (with an average age of 22 y) by correlating immunohistochemistry with specific anti-MHC monoclonal antibodies, myofibrillar ATPase (mATPase) histochemistry and in situ hybridisation with probes specific for MHC beta-slow, MHC-IIA and MHC-IIX. The characterisation of a large number of individual fibres was compared and correlated on a fibre-to-fibre basis. The panel of monoclonal antibodies used in the study allowed classification of human skeletal muscle fibres into 5 categories according to the MHC isoform they express at the protein level, types I, I+IIA, IIA, IIAX and IIX. Hybrid fibres coexpressing two isoforms represented a considerable proportion of the fibre composition (about 14%) and were clearly underestimated by mATPase histochemistry. For a very high percentage of fibres there was a precise correspondence between the MHC protein isoforms and mRNA transcripts. The integrated methods used demonstrate a high degree of precision of the immunohistochemical procedure used for the identification and quantification of human skeletal muscle fibre types. The monoclonal antibody S5-8H2 is particularly useful for identifying hybrid IIAX fibres. This protocol offers new prospects for muscle fibre classification in human experimental studies. SN - 0021-8782 UR - https://www.unboundmedicine.com/medline/citation/11554510/Immunolabelling_histochemistry_and_in_situ_hybridisation_in_human_skeletal_muscle_fibres_to_detect_myosin_heavy_chain_expression_at_the_protein_and_mRNA_level_ L2 - https://onlinelibrary.wiley.com/resolve/openurl?genre=article&sid=nlm:pubmed&issn=0021-8782&date=2001&volume=199&issue=Pt 3&spage=329 DB - PRIME DP - Unbound Medicine ER -