Fiber type specific distribution of stress proteins in rat skeletal muscle / 体力科学

ABSTRACT

To determine whether fiber type-specific expression of heat shock protein (HSP, or stress protein) occurs in unstressed rat skeletal muscle, the medial gastrocnemius of adult female Sprague-Dawley rats was subjected to immunohistochemical analysis. Antibodies against 5 types of anti-myosin heavy chain (MHC) were used to classify the type of fibers, and 2 types of anti-HSP antibodies were employed to analyze the fiber type-specific expression.<BR>Serial cross-sections of 10 μm thick cut by a cryostat were incubated with primary anti-MHC or anti-HSP 60 and 72 antibodies, followed by biotinylated secondary anti-mouse antibodies, and avidin-biotin complex solution. A peroxidase DAB substrate kit (Vector SK-4100) or BCIP/NBT solution was used to visualize the immunoreaction of each fiber type.<BR>By using the 5 types of anti-MHC antibodies, fibers were classified into 4 types slow-type I, fasttypes IIA, IIX, and IIB. Anti-HSP 72 antibody reacted with many, but not all, type I and IIA fibers, whereas anti-HSP 60 antibody reacted specifically with type I fibers. Neither type IIX nor IIB fibers showed immunoreactivity with anti-HSP 60 or 72 antibodies. These results suggest that the expression of HSP 60 protein is related to that of type I MHC, and that the expression of HSP 72 protein may be related to that of types I and ha MHC, in unstressed rat skeletal muscle.