Association of soluble guanylate cyclase with the sarcolemma of mammalian skeletal muscle fibers.
Previous investigations have shown that NO-producing nitric oxide synthase (NOS)-1 and CO-generating heme oxygenase (HO-2) are associated with the sarcolemma of skeletal muscle fibers in many mammalian species. Despite numerous roles ascribed to NO and possibly also CO in skeletal muscle, a specific receptor for both gases has hitherto not been found in myofibers. Therefore, in the present work the appearance of the alpha1, beta1 and beta2 subunits of soluble guanylate cyclase (sGC), the most commonly known receptor for NO and potentially also CO, was analysed in mammalian skeletal muscles using immunoblotting and immunohistochemistry. Immunoblotting with an antibody against the beta1 subunit of sGC revealed a band of 70 kDa corresponding to the molecular weight of this protein. Immunohistochemistry with antibodies against the alpha1, beta1 and beta2 sGC subunits showed that the larger part of positivity was present in the sarcolemma region of skeletal muscle fibers and colocalized with NOS-1 mainly in type II myofibers and with HO-2 in type I and type II myofibers. For the first time, sarcolemmal association of sGC and its colocalization with NOS-1 generating the sGC-activator NO and with HO-2 producing the potential sGC upregulator CO have been demonstrated in the present study. These results enable a better understanding of the role of NO and CO in myofibers and suggest a so far unknown molecular mechanism for the interaction of sGC with the sarcolemma.
Feussner
M
M
Department of Anatomy II, University Clinic Benjamin Franklin, Free University of Berlin, Germany.
Richter
H
H
Baum
O
O
Gossrau
R
R
D000818:Animals; D006224:Cricetinae; D005260:Female; D006162:Guanylate Cyclase; D006168:Guinea Pigs; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000201:enzymology*; D018482:Muscle, Skeletal / Q000201:enzymology*; D019001:Nitric Oxide Synthase / Q000032:analysis / Q000737:chemistry; D052248:Nitric Oxide Synthase Type I; D021122:Protein Subunits; D051381:Rats; D017208:Rats, Wistar; D018160:Receptors, Cytoplasmic and Nuclear / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D012508:Sarcolemma / Q000201:enzymology* / Q000378:metabolism; D000071756:Soluble Guanylyl Cyclase
D021122:Protein Subunits; D018160:Receptors, Cytoplasmic and Nuclear; C496320:NOS1 protein, human; D019001:Nitric Oxide Synthase; D052248:Nitric Oxide Synthase Type I; C496321:Nos1 protein, mouse; C496322:Nos1 protein, rat; D006162:Guanylate Cyclase; D000071756:Soluble Guanylyl Cyclase