Effects of fixation and preservation conditions on immunohistochemical profiles of the skeletal muscle fibers in Japanese macaques

Abstract

Effects of fixation and preservation conditions of muscle tissues on immunohistochemical profiles are investigated. Samples of the hind limb and epaxial muscles were removed from 4 adult female Japanese macaques (Macaca fuscata) fixated with 10% formalin and preserved in the same solution under different conditions for 6 months to 4 years and 6 months. Sections were stained with indirect immunofluorescence and avidin-biotin peroxidase complex methods using an antibody against fast myosin (Mouse Monoclonal Anti-skeletal Myosin-Fast, clone MY-32, Sigma) as a primary antibody. Clear responses to the antibody were demonstrated in the samples from the specimens fixated by injection or immersion with 10% formalin and preserved in the same solution for 6 months to 1 year and 6 months. Distribution patterns of the fibers reacting to the antibody coincided with that of the fast twitch fibers determined using enzyme-histochemical techniques in these samples. Clear responses to the antibody were not demonstrated in the samples from the specimen repeatedly rinsed in water for gross anatomical dissections during the preservation period. The results of this study warrant applications of immunohistochemical techniques to the study of fiber type composition in muscle samples from specimens fixated with formalin and preserved in the same solution for a long term.