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Irregular Costameres Represent Nitric Oxide Synthase-1-positive Sarcolemma Invaginations Enriched in Contracted Skeletal Muscle Fibres

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Abstract

NADPH diaphorase histochemistry and NOS-1 immunohistochemistry on 60 μm thick frozen sections of rat extensor digitorum longus muscles led to the detection of prominent rings clearly encompassing the surface of the muscle fibres. These so far unknown costameres were usually found as doublets flanking a space of about 2 μm width. Because these costameric doublets did not appear in regular periods, we designate them irregular costameres to discriminate them from regular ones with a 1 μm periodicity overlying Z-discs and M-lines. Irregular costameres were thicker than the regular ones and free of intercostameres. Immunohistochemistry demonstrated that NOS-1 was co-localized with integral β-dystroglycan, α-sarcoglycan) and peripheral (caveolin-3, dystrophin) members of the enlarged dystrophin complex in the irregular costameres but not with non-sarcolemmal organized proteins (myosin heavy chain, α-actinin, desmin and sarcoplasmic reticulum-located Ca2+-dependent ATPase-1). Invaginations of the sarcolemma to form irregular costameres were observed. In teased myofibres the sarcolemma between two following irregular costameres was ballooned, while the irregular costameres themselves clamped the fibres together. Finally, the number of detectable irregular costameres was significantly increased in maximally contracted extensor digitorum longus muscles generated by electric stimulation but decreased in mechanically stretched ones. Combining these observations, we hypothesize that irregular costameres belong to a reserve zone for the sarcolemma necessary for the contraction/relaxation cycle in myofibres.

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Baum, O., Planitzer, G., Richter, H. et al. Irregular Costameres Represent Nitric Oxide Synthase-1-positive Sarcolemma Invaginations Enriched in Contracted Skeletal Muscle Fibres. Histochem J 32, 743–751 (2000). https://doi.org/10.1023/A:1004153111532

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