Immunohistochemical Differentiation of Fiber Types in Human Skeletal Muscle Using Monoclonal Antibodies to Slow and Fast Isoforms of Troponin I Subunit

Abstract

The cDNA sequence of troponin I (TnI), one of the subunits of the skeletal muscle regulatory protein, differs between slow-twitch muscle and fast-twitch muscle. We prepared monoclonal antibodies td the slow and fast isoforms of human TnI for the purpose of differentiating muscle fiber types in human neuromuscular disorders. Slow TnI antibody was labeled with tetramethylrhodamine isothiocyanate while fast TnI antibody was labeled with fluorescein isothiocyanate; then these two antibodies were mixed. This mixture was then used to stain biopsied muscle from patients with neuromuscular disorders. It was possible to differentiate muscle fibers into slow, fast and intermediate fibers having various contents of slow and fast TnI. In tissue composed of small muscle fibers, this method facilitated differentiation of types of muscle fibers by allowing staining of only a single section. The usefulness of our technique using slow and fast TnI antibodies is discussed in comparison with ATPase staining. Because our staining method can distinguish slow and fast fiber components, it is useful for clinical application.