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Immunolocalization of ornithine decarboxylase in rat heart atria
U R Tipnis1, A L Steiner, C Skiera
1Department of Pharmacology, University of Texas Medical School, Houston 77225.
Abstract:
Mammalian atrial myocytes secrete a potent natriuretic and diuretic factor, atrial natriuretic peptide (ANP), in response to volume expansion or elevations of right atrial pressure. ANP is synthesized in the atrial atrial myocytes and stored in cytoplasmic secretory granules. Ornithine decarboxylase (ODC) is the initial rate-limiting enzyme in the biosynthesis of polyamines, organic cations which are essential for various aspects of DNA, RNA and protein synthesis, structure, and function. The enzyme has been reported to be localized predominantly in the cytoplasm. A polyclonal antibody elicited against ODC was used to analyse the intracellular distribution of the enzyme protein within atrial myocytes at the ultrastructural level through the use of a post-embedding immunogold technique. In control animals, the density of gold particles associated with the atrial granules was seven to 30-fold higher than that detected in association with other subcellular structures. Administration of Isoproterenol increased atrial of Isoproterenol increased atrial ODC activity 18-fold and the density of the immunolabelling of the atrial myocytes five-fold. Statistically significant increases in the density of labelling after stimulation occurred in association with the atrial granules and the nucleus. After isoproterenol stimulation, 60% of the immunodetectable ODC protein in the atrial myocyte was found in association with the atrial granules. The atrial granules were demonstrated to contain ANP by immunocytochemical analysis of adjacent sections.