B. J. Whitehouse

Oxytocin and arginine vasopressin stimulate steroid secretion by the isolated perfused rat adrenal gland

Using the intact isolated perfused rat adrenal preparation we have shown for the first time a direct effect of oxytocin on adrenocortical steroid secretion. Oxytocin specifically stimulated aldosterone secretion in a dose-dependent manner with a threshold dose of 1 pmol. Arginine vasopressin was also shown to be a potent stimulus to aldosterone secretion and was additionally found to stimulate inner zone function. Using superfused adrenal cells, the effects of arginine vasopressin were only seen at 10,000 times higher doses than were effective in the intact perfused gland, and oxytocin had no effect at any dose. These results reinforce the hypothesis that tissue integrity is essential for full expression of steroidogenic control mechanisms. We conclude that oxytocin and vasopressin may play a role in the control of steroidogenesis.

Effect of arginine vasopressin and oxytocin on acetylcholine-stimulation of corticosteroid and catecholamine secretion from the rat adrenal gland perfused in situ

The effects of acetylcholine, arginine vasopressin (AVP) and oxytocin (OXT) on both catecholamine and steroid secretion have been investigated using the isolated rat adrenal gland perfused in situ. Significant stimulation of steroid (aldosterone and corticosterone) secretion occurred with 1 mumol/l acetylcholine; the ED50 was approximately 20-fold higher (circa 20 mumol/l) than that for catecholamine secretion. The highest concentration of acetylcholine used (100 mumol/l) stimulated aldosterone secretion eight-fold; corticosterone secretion four-fold; noradrenaline and adrenaline secretion three-fold. AVP at 100 nmol/l but not at 1 nmol/l significantly stimulated the secretion of both steroids and catecholamines. OXT had no significant effect on corticosteroid or catecholamine secretion at either concentration. The effects on aldosterone secretion of simultaneous administration of acetylcholine and AVP were additive. No similar effect was seen on corticosterone or catecholamine secretion where the degree of stimulation was the same as for acetylcholine alone. OXT (100 nmol/l) inhibited acetylcholine-stimulated aldosterone secretion but had no effect on acetylcholine-stimulated catecholamine secretion. Carbachol was equipotent with acetylcholine in stimulating steroid secretion from the perfused gland. Our results support the hypothesis that acetylcholine may play a role in the control of steroid secretion by the rat adrenal cortex. They fail to support a role for AVP and OXT in modulating catecholamine secretion by the adrenal medulla except at high concentrations.

The biosynthesis of aldosterone

The cellular mechanisms for aldosterone biosynthesis are incompletely understood. Although the enzymes involved are now well characterized, the dynamics of aldosterone secretion in a variety of rat adrenal preparations are not consistent with the concept that freshly synthesized corticosterone is an important intermediate. In whole glomerulosa tissue preparations, aldosterone is more readily formed from endogenous precursors than from an added radioactive precursor, such as [3H]pregnenolone, and in the in situ perfused gland preparation, aldosterone responses to stimulation, for example by ACTH, are significantly more rapid than those of corticosterone, suggesting a tissue source of steroid substrate for aldosterone production other than corticosterone. The only steroid which is stored in rat adrenal glomerulosa tissue to any extent is 18-hydroxydeoxycorticosterone (18-OH-DOC), and this pool has been located in plasma membrane fractions. It is lost on preparation of collagenase dispersed glomerulosa cells. Since dispersed glomerulosa cell preparations produce significantly less aldosterone, relative to corticosterone, than incubated intact whole glomerulosa, it is plausible that this tissue pool (which is not found in the inner zones) is the immediate precursor for aldosterone formation. Further evidence shows that trypsin, which stimulates aldosterone (and 18-hydroxycorticosterone) production in rat intact glomerulosa tissue, but not in dispersed cells, stimulates translocation of protein kinase C to the plasma membrane. It is plausible that one function of protein kinase C in the rat adrenal zona glomerulosa is to mobilize membrane sequestered 18-OH-DOC for conversion to aldosterone.

Control of aldosterone secretion in zona glomerulosa cell suspensions and in the perfused adrenal gland of the rat.

The secretion of aldosterone and its responses to stimulation have been studied in rat adrenal zona glomerulosa tissue incubated as intact capsules or as collagenase-dispersed cell suspensions, and in intact perfused rat adrenal glands. Several differences are apparent in the functions of the various preparations. Aldosterone secretion rates are similar in incubated intact capsules and in the perfused gland. Relative to corticosterone, lower yields of aldosterone are obtained in dispersed glomerulosa cell in vitro. This may be related to the loss in the dispersed cells of a pool of tissue steroid (aldosterone or a precursor) which is revealed only in intact tissue incubations by trypsin stimulation of aldosterone secretion. Trypsin-released aldosterone is increased by prior dietary sodium restriction. In addition, differences occur in the responses of dispersed cells and perfused glands to stimulation. Perfused glands from animals on a normal diet are less sensitive to stimulation by ACTH or alpha-MSH, but more sensitive than dispersed cells to angiotensin II amide. In the perfused gland, sensitivity of response (lowest effective concentration) to all three stimulants is increased by prior dietary sodium restriction, in contrast to dispersed cells in which increased sensitivity has been reported only to alpha-MSH. The perfused gland is particularly sensitive to angiotensin II amide, and a bolus administration of 1 amol gives significant stimulation in glands from animals on low sodium intake. Electrical (field) stimulation or dopamine administration at 10(-6) mol/l (which is ineffective in dispersed cells) both depress aldosterone secretion by the perfused gland. The data suggest that the sequestered pool of steroid is utilized in the perfused gland for aldosterone secretion. They furthermore suggest that in the intact gland there are mechanisms, which involve neural components, for intraglandular regulation of aldosterone secretion, which are lost in dispersed cells in vitro. Such mechanisms may be involved in sensitivity increases in sodium depletion.