Plasma hyperosmolality increases G protein and 3',5'-cyclic adenosine monophosphate synthesis in the paraventricular and supraoptic nuclei.

Hyperosmotic stimuli produce profound changes in cellular morphology and biosynthetic activities within the hypothalamic paraventricular and supraoptic nuclei (SON) of the rat. The mechanisms by which osmoreceptive signals are transduced within these nuclei are poorly understood. We examined several components of the cAMP-associated second messenger system after giving rats 2% saline to drink for one week, a strong hyperosmotic stimulus. We found that mRNA levels for both the stimulatory and inhibitory guanine-nucleotide binding protein alpha-subunits were increased in the paraventricular nucleus and SON. In the SON, these changes were accompanied by increased basal cAMP levels, cholera toxin-stimulated adenylate cyclase, and Gs alpha. Our results suggest that Gs alpha levels are not saturated with respect to adenylate cyclase coupling and that osmoreception activates the cAMP second messenger system.

Cytochrome b561 can be detected in many neuroendocrine tissues using a specific monoclonal antibody.

CG7 is a monoclonal antibody that recognizes cytochrome b561, a major protein component of adrenal medullary chromaffin granules. Immunocytochemical studies using this antibody have demonstrated that cytochrome b561 is present in many neural and endocrine tissues and that its distribution is correlated with the presence of either catecholamines or amidated peptides in the tissue. These tissues include neuronal cell bodies and/or fibers in the gut, blood vessels, retina, and posterior pituitary, endocrine cells of the gut, anterior and intermediate lobes of the pituitary, heart muscle, and all adrenal medullary chromaffin cells. The discovery of cytochrome b561 in many neuropeptide-containing tissues regardless of the presence of catecholamines is consistent with a general role for cytochrome b561 as a secretory granule membrane electron carrier. Its expression may be linked to an ascorbic acid requirement by both catecholamine and neuropeptide biosynthetic enzymes.

Cell-specific expression of the rat oxytocin gene in transgenic mice.

Abstract The genes for the posterior pituitary hormones oxytocin (OT) and vasopressin (VP) are expressed in magnocellular neurons of the hypothalamus. Previous attempts to obtain cell-specific expression of OT and VP transgenes in mice have been unsuccessful using constructs containing only the OT or VP genes. As the endogenous genes are located near each other on the same chromosome, we investigated four transgenic mouse lines incorporating a single 5.2 k bp construct of rat genomic deoxyribonucleic acid with 1.63 k bp and 3.55 k bp of the OT and VP genes, respectively. Rat OT transcripts were detected only in transgenic mouse OT neurons of the paraventricular and supraoptic nuclei in two lines. The levels of endogenous and transgene OT transcripts increased in response to 10 days of lactation. Furthermore, rat OT-associated neurophysin immunoreactivity was detected in magnocellular cell bodies as well as in the posterior pituitary of the transgenic but not the control mice. Cell-specific rat VP expression was not detected in these transgenic mice.