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Journal of Histochemistry and Cytochemistry, Vol. 50, 353-364, March 2002, Copyright © 2002, The Histochemical Society, Inc.

ARTICLE

Quantifying the Temporospatial Expression of Postnatal Porcine Skeletal Myosin Heavy Chain Genes

Nuno da Costaa, Ross Blackleya, Hadi Alzuherria, and Kin-Chow Changa
a Veterinary Molecular Medicine Laboratory, Department of Veterinary Pathology, University of Glasgow, Glasgow, Scotland

Correspondence to: Kin-Chow Chang, Veterinary Molecular Medicine Laboratory, Dept. of Veterinary Pathology, Univ. of Glasgow, Bearsden Road, G61 1QH Glasgow, Scotland. E-mail: k.chang@vet.gla.ac.uk

Postnatal skeletal muscle fiber type is commonly defined by one of four major myosin heavy chain (MyHC) gene isoforms (slow/I, 2a, 2x, and 2b) that are expressed. We report on the novel use of combined TaqMan quantitative real-time RT-PCR and image analysis of serial porcine muscle sections, subjected to in situ hybridization (ISH) and immunocytochemistry (IHC), to quantify the mRNA expression of each MyHC isoform within its corresponding fiber type, termed relative fiber type-restricted expression. This versatile approach will allow quantitative temporospatial comparisons of each MyHC isoform among muscles from the same or different individuals. Using this approach on porcine skeletal muscles, we found that the relative fiber type-restricted expression of each postnatal MyHC gene showed wide spatial and temporal variation within a given muscle and between muscles. Marked differences were also observed among pig breeds. Notably, of the four postnatal MyHC isoforms, the 2a MyHC gene showed the highest relative fiber type-restricted expression in each muscle examined, regardless of age, breed, or muscle type. This suggests that although 2a fibers are a minor fiber type, they may be disproportionately more important as a determinant of overall muscle function than was previously believed. (J Histochem Cytochem 50:353–364, 2002)

Key Words: quantitative real-time PCR, myosin heavy chain, fiber type, temporospatial expression, porcine


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