An immunofluorescent analysis of lectin binding to normal and regenerating skeletal muscle of rat

Abstract

Ten different fluorescein-conjugated lectins of various known sugar specificities were used to study cell surface glycoconjugates of normal and regenerating rat skeletal muscle. In normal muscle, Canavalia ensiformis agglutinin, Triticum vulgaris agglutinin (wheat germ agglutinin, WGA), Ricinus communis agglutinin-I, and Maclura pomifera agglutinin bound strongly to the endomysial region of the myofibers. No binding was observed in the cytoplasm of the myofibers. Other lectins (Dolichos biflorus agglutinin, Griffonia simplifolia agglutinin I and II, Ulex europaeus agglutinin, Arachis hypogaea agglutinin, Glycine max agglutinin) bound very poorly or not at all in the normal muscle. Skeletal muscle degeneration and regeneration was induced by autotransplantation of the extensor digitorum longus muscle. In the transplanted muscles, the endomysial lectin binding of the degenerating myofibers became weak and lacked continuity, indicating a breakdown of the endomysium. Of all the lectins tested, only WGA revealed intense binding in the myogenic zone of regenerating muscle. As the regeneration progressed, this WGA binding became restricted to the new endomysium. In completely regenerated muscle, binding of all lectins was similar to that seen in normal muscle. The specific binding of WGA to the myogenic zone may be important in identifying factors or glycoconjugates, which constitute a favorable environment for skeletal muscle regeneration.

Application of WGA lectin staining for visualization of the connective tissue in skeletal muscle, bone, and ligament/tendon studies

“…Compared to the desmin staining that was lost during early stages of necrosis, lectin binding to the periphery of the muscle fibers was preserved (Helliwell,1988). Gulati and Zalewski (1985) also showed that of four lectins studied only WGA had intense binding in the myogenic zone of regenerating muscle after autotransplantation.…”

Lectin Histochemistry on Glycoconjugates of the Epidermis and Dermal Glands of Xenopus laevis (Daudin, 1802)

“…Such discrepancies, previously emphasized by several authors (Holthofer 1983;Nemanic et af. 1983;Schulte & Spicer 1983;Gulati & Zalewski 1985;Meiniel et a f . 1988;Truong et af.…”

A method for preparing skeletal muscle fiber basal laminae

“…Although some immunocytochemical investigations on the BL of degenerating and regenerating muscle fibers have been performed (Gulati et al, 1983;Gulati, 1985;Gulati and Zalewski, 1985), further analysis of the properties of muscle fiber BL during degeneration and regeneration has been hampered by the lack of suitable preparative methods. In the current study we describe a method for the generation of acellular muscle samples which exhibit numerous empty tubes of BL which are available for analysis by light microscopy, TEM or SEM.…”