Atrial Natriuretic Peptide Secretion from Fetal Rat Diencephalon in Culture

The presence of a distinct brain pool of the atrial natriuretic peptides (ANP) has been established. To determine the molecular forms and regulation of secretion of ANP, we studied fetal rat diencephalic neurons and glia in primary culture. ANP immunoreactivity determined by radioimmunoassay was found only in the neuron predominant cultures. The neurons contained mainly ANP (103-126) and less ANP (102-126), but secreted only ANP (103-126) into the medium after potassium and glutamate-dependent depolarization. Little, if any, ANP (99-126), the predominant form which circulates in plasma and originates from the heart, was secreted. The ability of potassium and glutamate to cause a mean 50% increase of ANP secretion above baseline was abolished after deleting calcium chloride from the medium. In contrast, hypo- or hyperosmolarity or increased sodium content in the incubation medium did not influence ANP secretion. These studies indicate that regulative secretion of ANP occurs from primary cultures of predominantly diencephalic neurons, probably accounting for the high concentrations of these peptides in this area of the brain. The forms of ANP contained within the cells and secreted after depolarization are different from ANP secreted from neonatal rat atrial myocytes. In contrast to myocytes, varying sodium or osmolarity did not cause ANP secretion. We postulate that influences on ANP production/secretion in the brain may be distinct from the heart.