Histochemical and molecular determination of fiber types in chemically skinned single equine skeletal muscle fibers.

Abstract

Until now, there has been no reliable method for histochemical determination of fiber types of single skinned muscle fibers. The major problem arises from the fact that most histochemical techniques use cross-sections of a large group of fibers and compare a given fiber with those surrounding it. This is not possible with a single skinned fiber which has been separated from a bundle to be used for mechanical analysis. A further problem is that the skinning procedure itself may alter the staining pattern. We have developed a procedure by which multiple cross-sections of single skinned fibers can be exposed to various histochemical reactions and the staining patterns compared on the same slide to those of frozen muscle and skinned bundles. By this procedure, three fiber types were distinguished by both Ca2+-ATPase and SDH reactions. The fiber typings determined from both enzyme systems correlated well with each other. Although we were able to differentiate only between slow and fast fibers by SDS-PAGE, these results corroborated the histochemical classification. This procedure will clearly be useful in skinned single muscle fiber mechanics experiments performed to determine functional differences among fiber types.

Immunohistochemical Classification of Skeletal Muscle Fibers.

“…Some other proteins such as myosin light chain, troponin, actin, sarcoplasmic (endoplasmic) reticulum Cat+-ATPase (SERCA), and parvalbumin contribute to the identification of specific fiber types and have different isoform expressions between slow-and fast-twitch fibers [56,69,71,74,91,95]. The functional diversity of the skeletal muscle is, however, most adequately represented by the MHC isoform distribution of the muscle.…”

The influence of temperature on mechanics of red muscle in carp.

“…The bundles were then gently blotted with soft filter paper, immersed in a gelatin supportive medium (as in Sosnicki, Lutz, Rome & Goble, 1989) and then rapidly frozen in isopentane cooled with liquid nitrogen while still connected to the servomotor. Serial cross-sections 8 Jam thick throughout the entire length of the bundles were obtained with a cryostat (Tissue-Tek, Ames, Elkhart, IN, USA).…”

Muscle Fiber Population and Biochemical Properties of Whole Body Muscles in Thoroughbred Horses

“…It is well known that muscles of the Thoroughbred horse are composed of Type I, Type IIa, and Type IIx fibers (Billeter et al, 1988;Sosnicki et al, 1989;Rivero et al, 1996;Serrano et al, 1996). Furthermore, the development of immunohistochemical techniques has permitted identification of hybrid muscle fibers expressing two types of myosin heavy chain (MHC) isoforms (Linnane et al, 1999;Rivero et al, 1999;Quiroz-Rothe and Rivero, 2001).…”