Ventricular and atrial myocytes of newborn rats synthesise and secrete atrial natriuretic peptide in culture: Light-and electron-microscopical localisation and chromatographic examination of stored and secreted molecular forms |
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Authors: | C. J. S. Hassall J. Wharton S. Gulbenkian J. V. Anderson J. Frater D. J. Bailey A. Merighi S. R. Bloom J. M. Polak G. Burnstock |
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Affiliation: | (1) Department of Anatomy and Developmental Biology and Centre for Neuroscience, University College London, London, UK;(2) Departments of Histochemistry and Medicine, Royal Postgraduate Medical School, London, UK;(3) Department of Anatomy and Developmental Biology and Centre for Neuroscience, University College London, Gower Street, WC1E 6BT London, UK |
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Abstract: | Summary We have demonstrated that atrial natriuretic peptide-like immunoreactivity is stored and secreted by ventricular and atrial myocytes in dissociated cell culture preparations from the heart of newborn rat. Culture preparations were maintained in either foetal calf serum-supplemented medium 199 or in hormone-supplemented, serum-free medium 199. The presence of atrial natriuretic peptidelike immunoreactivity in the cultured myocytes was demonstrated at both light-and electron-microscopical levels. Release of atrial natriuretic peptide-like immunoreactivity into the culture medium was measured by radioimmunoassay; molecular forms of the stored and secreted peptide were determined by gel column chromatography. The atrial natriuretic peptide-like immunoreactivity of cultured atrial and ventricular myocytes was concentrated in the perinuclear cytoplasm and was localised to electron-dense secretory granules. The number of immunoreactive ventricular myocytes and the intensity of their immunofluorescence changed with time in culture and was higher in cultures in foetal calf serum-supplemented medium than in serum-free medium. Gamma-atrial natriuretic peptide was stored and released by cultured atrial and ventricular myocytes, but was broken down to alpha-atrial natriuretic peptide in the growth medium. This process was foetal calf serum-independent, since it occurred in both the media used, indicating that cardiac myocytes in culture may release a factor that cleaves gamma-atrial natriuretic peptide to form alphaatrial natriuretic peptide. |
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Keywords: | Atrial natriuretic peptide Ventricular myocytes Atrial myocytes Cell culture Secretion Rat |
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