The nitric oxide synthase-1 and nitric oxide synthase-3/nitric oxide signalling systems in the heart of wild type mice and mouse mutants.

Abstract

Recently, we have shown that nitric oxide synthase-1 (NOS-1) and thus its product NO are present in the sarcolemma region of a subpopulation of atrial cardiomyocytes in the rat heart. In order to find out whether this newly discovered sarcolemma-associated NOS/NO system represents a general signalling mechanism in the murine rodent heart and whether its properties are comparable to those in skeletal muscle fibres, immunohistochemical and catalytic histochemical methods (including image analysis) were applied to the heart and extensor digitorum longus (EDL) and tongue muscles of wild type and mutant mice. In different strains of wild type mice and NOS-3 knockouts, urea-resistant (and therefore specific) NOS NADPH diaphorase histochemistry and NOS-1 immunohistochemistry revealed that NOS-1 activity and protein were present in the sarcolemma region of a subpopulation of atrial and ventricular working cardiomyocytes, but not in those of the impulse conducting system. Using image analysis, NOS-1 showed similar activities in the sarcolemma region of cardiomyocytes and in EDL type I myofibres. In mdx and NOS-1 knockout mice, NOS-1 was absent from the sarcolemma region of atrial and ventricular cardiomyocytes and of EDL and tongue muscle fibres, whereas NOS-1 was present in the hearts of NOS-3 knockouts. Atrial natriuretic peptide immunohistochemistry identified part of the atrial NOS-1-expressing cardiomyocytes as myoendocrine cells. In mdx mice as well as in NOS-1 - and NOS-3-deficient animals, the peptide was found in greater abundance than in wild type mice. These data suggest that NOS-1 is expressed in a subpopulation of working cardiomyocytes in the murine rodent heart, that the myoendocrine cells may be negatively modulated by NOS-1 - and NOS-3-produced NO, and that the anchoring mechanisms for NOS-1 in these cells (i.e. their confinement to the sarcolemma region) are comparable to those in skeletal muscle fibres.