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Oxytocin (OT), historically known for its actions on the reproductive system, may also contribute to the regulation of cardiovascular and water-electrolyte homeostasis. OT is produced in the supraoptic and paraventricular nuclei of the hypothalamus and released into plasma from neural terminals of the posterior pituitary, however, many studies have identified extra-cerebral sites of OT production, including the heart and vascular endothelium. Activation of its receptors on endothelial cells, as well as in hypothalamic/hypophyseal and cardiac systems, may result in nitric oxide (NO) production. We aimed to verify the role of NO in the regulation of OT-stimulated atrial natriuretic peptide (ANP) secretion in primary culture of mouse embryonic cardiomyocytes. For this, we used hearts from Balb C mouse embryos, 19 to 21 days old intrauterine, were isolated and cultured for assays with OT and other interfering substances in the synthesis of NO and cGMP its second messenger.