Expression of provasopressin gene during ontogeny in the hypothalamus of developing mice

Abstract

We investigated the ontogeny of provasopressin gene expression in neurosecretory neurons of the supraoptic and paraventricular nuclei of developing mice by semi-quantitative in situ hybridization and immunohistochemical techniques in combination with stereometry of vasopressin-immunoreactive neurons. Provasopressin mRNA was detected in paraffin sections using a mixture of radiolabeled synthetic oligonucleotide probes complementary to the mRNA loci encoding vasopressin (2-9) and vasopressin neurophysin (1-8). Vasopressin immunoreactivity was located with a polyclonal anti-vasopressin antiserum and a monoclonal anti-vasopressin-neurophysin antibody either with or without enhancing technique for the diaminobenzidine reaction. Autoradiographic hybridization signals that indicate the localization of provasopressin mRNA were first detected on embryonic day 15 in the supraoptic nucleus and embryonic day 18 in the paraventricular nucleus. Vasopressin immunoreactivity was first found in the median eminence on embryonic day 14, and then in the supraoptic and paraventricular nuclei on embryonic days 15 and 16, respectively. The provasopressin mRNA levels were markedly increased in both the supraoptic and the paraventricular nuclei just after birth. The immunoreactivity of vasopressin neurons was drastically decreased in both nuclei on postnatal days 1 and 2, suggesting marked vasopressin release in the neonates. Cross-sectional areas of vasopressin-immunoreactive somata and their cell nuclei gradually increased in both the supraoptic and the paraventricular nuclei during the perinatal period by day 5, and then attained adult size between days 10 and 20. During this phase, the level of provasopressin mRNA remained low compared with that in the adult magnocellular neurosecretory cells. These results indicate that the expression of provasopressin gene is markedly increased in both the supraoptic and the paraventricular nuclei soon after birth. Secretory activity of vasopressin neurons is elevated in neonatal mice. Vasopressin may have an important osmoregulatory role in neonatal mice undergoing drastic changes in water metabolism following birth.