Effect of ACTH or zinc treatment on plasma aldosterone and corticosterone levels and on the in vitro steroid output from adrenocortical cells

1982 ◽  
Vol 60 (11) ◽  
pp. 1058-1064 ◽  
Author(s):  
Nicole Payet ◽  
Jean-Guy Lehoux
Keyword(s):  
Day Treatment ◽  
Zona Glomerulosa ◽  
Plasma Aldosterone ◽  
Female Rats ◽  
Zinc Hydroxide ◽  
Elevated Plasma ◽  
Adrenocortical Cells ◽  
Glomerulosa Cells ◽  
Zona Glomerulosa Cells

We have studied the effect of in vivo treatment with two forms of ACTH (Synacthen and Duracton) and of zinc hydroxide on plasma corticosteroid levels from adult Long Evans female rats. The corticosteroid output by isolated zona glomerulosa cells in vitro was also studied. Dose–response experiments showed that, after 2 days of treatment, Synacthen caused a 2.5- and 25.9-fold increase in plasma aldosterone and corticosterone levels, respectively, while maximal increases of 1.7- and 5.6-fold were obtained following treatment with Duracton. In contrast to elevated plasma steroid levels, the basal aldosterone and corticosterone output by isolated zona glomerulosa cells was significantly decreased for all doses of Synacthen administered. The 8 IU/day treatment with Synacthen produced an 86% diminution of aldosterone output while a treatment of 32 IU/day with Duracton gave only a 48.8% decrease. Concomitantly the Synacthen treatment provoked a high mitotic response in the zona glomerulosa cells (fivefold over control). A 2-week treatment with Synacthen resulted in elevated plasma aldosterone and corticosterone levels and produced a 92% diminution of aldosterone output by isolated zona glomerulosa cells. The aldosterone and corticosterone output from these cells was not enhanced by the addition of ACTH in the incubation media; zona glomerulosa cells of animals treated with Synacthen were no longer responsive to ACTH stimulation in vitro. The effect of a 2-day treatment with zinc hydroxide on plasma aldosterone and corticosterone levels and on steroid output by isolated adrenocortical cells was different from that of Synacthen. This meant that zinc was not the active principle of the Synacthen preparation. Our results indicate that long-term treatment with ACTH provokes profound functional changes at the adrenocortical zona glomerulosa level.

scholarly journals Guinea pig adrenocortical cells: in vitro characterization of separated zonal cell types.

1982 ◽  
Vol 94 (2) ◽  
pp. 241-252 ◽  
Author(s):  
P Miao ◽  
V H Black
Keyword(s):  
Guinea Pig ◽  
Morphological Characteristics ◽  
Zona Glomerulosa ◽  
Equilibrium Density ◽  
Zona Fasciculata ◽  
Adrenocortical Cells ◽  
Steroid Production ◽  
Glomerulosa Cells ◽  
Zona Glomerulosa Cells

This paper reports a quick, relatively simple and reproducible technique for obtaining populations of zona fasciculata and zona glomerulosa cells up to 80-90% pure, which can be maintained in vitro for study of adrenocortical cell function. Isolated guinea pig adrenocortical cells were separated on a 1-28% bovine serum albumin/Ca++, Mg++-free buffer gradient (wt/vol at 4% increments) using equilibrium density centrifugation (570 g, 30 min). Over 60% of the 8 x 10(5) viable cells/adrenal obtained in the total isolate were recovered after separation. 80% of the zona glomerulosa cells were found in the lower three bands of the gradient. 78% of the zona fasciculata cells were found in the top three bands. Of the cells in the first two bands, 78-91% were zona fasciculata cells, whereas of the cells in the bottom two bands 92-95% were zona glomerulosa cells. The cells retained the morphological characteristics of cells in situ and could be maintained in vitro for periods up to 11 d. They produced a wide variety of steroids, cortisol, corticosterone, aldosterone, 11-beta-hydroxyandrostenedione, deoxycortisol, deoxycorticosterone, cortisone, 18-hydroxycorticosterone, and a product tentatively identified as dehydroepiandrosterone, and they responded to ACTH in a dose-responsive manner with enhanced levels of steroid output. Zona glomerulosa-enriched populations differed from zona fasciculata-enriched populations in their abundant production of aldosterone and in the pattern of steroid production. None of the cultures responded to angiotensin II (100 pg/ml) with increased steroid production.

Inhibition of aldosterone release by hypoxia in vitro: interaction with carbon monoxide

1994 ◽  
Vol 76 (2) ◽  
pp. 689-693 ◽  
Author(s):  
H. Raff ◽  
B. Jankowski
Keyword(s):  
Carbon Monoxide ◽  
Angiotensin Ii ◽  
Zona Glomerulosa ◽  
Michaelis Constant ◽  
Aldosterone Synthase ◽  
Aldosterone Production ◽  
Glomerulosa Cells ◽  
In Vitro Interaction ◽  
Zona Glomerulosa Cells

We have demonstrated that the aldosteronogenic pathway of the zona glomerulosa is unusually sensitive to modest changes in PO2 (Michaelis constant for O2 approximately 95 Torr). The current study evaluated the interaction of CO (the classic ligand for P-450 enzymes) and the decreases in O2 on aldosteronogenesis in vitro. Bovine adrenocortical zona glomerulosa cells were incubated for 2 h and stimulated with either adenosine 3′,5′-cyclic monophosphate (cAMP) or angiotensin II. Ten and 20% CO led to significant decreases in cAMP- and angiotensin II-stimulated aldosteronogenesis. The combination of 20% CO and moderate decreases in PO2 (from approximately 140 to approximately 100 Torr) led to an interactive decrease in aldosterone production. The conversion of corticosterone to aldosterone catalyzed by aldosterone synthase, which is the site of O2 sensitivity, was not significantly inhibited by CO. We conclude that the aldosterone pathway is not exceptionally sensitive to CO compared with other steroidogenic pathways. This observation suggests that the unique O2-sensitive properties of the aldosterone pathway located primarily within aldosterone synthase may not reside in its CO binding site (i.e., heme).

Inhibition of corticosteroid production by sodium pentobarbitone in rat adrenocortical preparations

1993 ◽  
Vol 136 (1) ◽  
pp. 75-83 ◽  
Author(s):  
B. J. Whitehouse ◽  
S. J. Purdy ◽  
D. R. E. Abayasekara
Keyword(s):  
Intact Animal ◽  
Zona Glomerulosa ◽  
Zona Fasciculata ◽  
Steroid Production ◽  
Adrenal Cells ◽  
Glomerulosa Cells ◽  
Pituitary Adrenal Axis ◽  
Zona Glomerulosa Cells ◽  
Stimulation Of

ABSTRACT It is possible that some of the effects of sodium pentobarbitone on the hypothalamo-pituitary-adrenal axis in the intact animal may be attributable to direct actions on the adrenal cortex. The effects of the barbiturate on steroid production by rat adrenal preparations in vitro have therefore been examined. In zona glomerulosa cells, pentobarbitone inhibited basal steroid production in a dose-related fashion. For aldosterone and corticosterone, the doses required for 50% inhibition of production (IC50) were 1·2 mmol pentobarbitone/l and 3·7 mmol/l respectively. Steroidogenesis was inhibited at lower levels of pentobarbitone in the presence of 1 nmol ACTH/l (IC50 = 0·5 mmol pentobarbitone/l for aldosterone and 2·2 mmol/l for corticosterone). In zona fasciculata/reticularis cells, production of corticosterone was similarly reduced with an IC50 of 2·8 mmol pentobarbitone/l for basal production and 1·3 mmol/l for ACTH-stimulated production. The dose-related increases in corticosterone production produced by ACTH (0·1–1000 pmol/l) or dibutyryl cyclic AMP (0·1–1·0 mmol/l) were also eliminated in the presence of 2 mmol pentobarbitone/l. The effects of pentobarbitone (1–4 mmol/l) on the production of pregnenolone and deoxycorticosterone (DOC) were also studied. In zona fasciculata/reticularis cells, the responses of both pregnenolone and DOC were bell-shaped with increases at 1 mmol pentobarbitone/l, which fell back to control levels at 4 mmol pentobarbitone/l. Stimulation of DOC, accompanied by decreases in aldosterone and corticosterone production, was also seen in zona glomerulosa cells at 1 mmol pentobarbitone/l. The effect of 1 mmol pentobarbitone/l on the conversion of 22(R)-hydroxycholesterol (5-cholestene-3β,22(R)-diol), pregnenolone, progesterone and DOC to corticosterone and aldosterone by zona glomerulosa preparations was studied. There was a comparable reduction in the conversion of these precursors (2 μmol/l) to aldosterone with yields decreased to 20–30% of those found in the absence of pentobarbitone. The dose required for 50% reduction of the conversion of progesterone (2 μmol/l) to aldosterone was 0·55 mmol pentobarbitone/l and for corticosterone the dose was 1·75 mmol pentobarbitone/l. The results obtained show that pentobarbitone is an effective inhibitor of corticosteroid biosynthesis in rat adrenal cells, and suggest that its effects are brought about by inhibition of cytochrome P450-mediated hydroxylations. Journal of Endocrinology (1993) 136, 75–83

scholarly journals Local production and action of adrenomedullin in the rat adrenal zona glomerulosa

1998 ◽  
Vol 156 (3) ◽  
pp. 477-484 ◽  
Author(s):  
S Kapas ◽  
A Martinez ◽  
F Cuttitta ◽  
JP Hinson
Keyword(s):  
In Situ Hybridization ◽  
Inositol Phosphate ◽  
Zona Glomerulosa ◽  
Binding Studies ◽  
Aldosterone Secretion ◽  
Glomerulosa Cells ◽  
Camp Formation ◽  
Zona Glomerulosa Cells

This study was designed to investigate the synthesis and action of adrenomedullin in the rat adrenal gland. The results obtained from in situ hybridization and immunocytochemical studies suggest that adrenomedullin is synthesized not only in the medulla, but also within the zona glomerulosa of the rat adrenal cortex. Findings from in situ hybridization and binding studies also suggested that specific adrenomedullin receptors are expressed in the zona glomerulosa, and that low levels are present in the inner zones of the cortex. The Kd of the zona glomerulosa adrenomedullin receptor (5.5 nmol/l) suggests that it may respond to locally produced adrenomedullin rather than circulating concentrations of the peptide, which are in a lower range. It was found that adrenomedullin acted on zona glomerulosa cells in vitro to stimulate aldosterone release and cAMP formation, but in this tissue did not stimulate inositol phosphate turnover. The effect of adrenomedullin on aldosterone secretion was significantly attenuated by a protein kinase A inhibitor, suggesting that cAMP mediates the effects of adrenomedullin on aldosterone secretion. Adrenomedullin did not significantly affect the response of zona glomerulosa cells to stimulation by either ACTH or angiotensin II. Adrenomedullin did not affect the release of catecholamines, either adrenaline or noradrenaline, by intact adrenal capsular tissue. These data suggest that both adrenomedullin and its specific receptor are expressed in the rat adrenal zona glomerulosa, leading to the hypothesis that adrenomedullin may have an autocrine/paracrine role in the regulation of the rat adrenal zona glomerulosa.

Potassium-induced aldosterone biosynthesis in cultured rat zona glomerulosa cells

1989 ◽  
Vol 256 (4) ◽  
pp. E475-E482
Author(s):  
J. Muller ◽  
M. Lauber ◽  
C. Schmid
Keyword(s):  
Molecular Mechanisms ◽  
Primary Cultures ◽  
Zona Glomerulosa ◽  
Zona Fasciculata ◽  
High Potassium ◽  
Primary Monolayer ◽  
Glomerulosa Cells ◽  
Potassium Restriction ◽  
Zona Glomerulosa Cells

Rat adrenal zona glomerulosa cells lost their ability to produce aldosterone from either endogenous precursors or added deoxycorticosterone within 2 days of primary monolayer culture in a medium with a potassium concentration of 6.3 mmol/l. The lost corticosterone methyl oxidase I and II activities were totally regenerated when the ambient potassium concentrations was raised to 31 mmol/l. The conversions of deoxycorticosterone to 18-hydroxycorticosterone and aldosterone were completely restored by culture in a high-potassium medium also in zona glomerulosa cells of rats in which aldosterone biosynthesis had been suppressed by potassium restriction and sodium loading. However, these conversions were not induced in zona fasciculata-reticularis cells. The induction of aldosterone biosynthesis was associated with the appearance of a mitochondrial 49,000 protein cross-reacting with an antibody raised against bovine adrenal cytochrome P-450(11) beta. Thus primary cultures of zona glomerulosa cells are promising models for studying in vitro the molecular mechanisms of long-term adaptation of aldosterone biosynthesis to sodium and potassium intake.

Dopamine increases interleukin 6 release and inhibits tumor necrosis factor release from rat adrenal zona glomerulosa cells in vitro

1996 ◽  
Vol 134 (5) ◽  
pp. 610-616 ◽  
Author(s):  
Paul K Ritchie ◽  
Marilyn Ashby ◽  
Heather H Knight ◽  
Allan M Judd
Keyword(s):  
Tumor Necrosis Factor ◽  
Interleukin 6 ◽  
Tumor Necrosis ◽  
Zona Glomerulosa ◽  
Adrenal Cells ◽  
Glomerulosa Cells ◽  
Necrosis Factor ◽  
Factor Release ◽  
Zona Glomerulosa Cells

Ritchie PK, Ashby M, Knight HH, Judd AM. Dopamine increases interleukin 6 release and inhibits tumor necrosis factor release from rat adrenal zona glomerulosa cells in vitro. Eur J Endocrinol 1996:134:610–6. ISSN 0804–4643 Interleukin 6 (IL-6) and tumor necrosis factor (TNF) are released from the zona glomerulosa of rat adrenal glands. The release of these cytokines from adrenal cells is regulated by interleukin 1β (IL-1β) and lipopolysaccharide (LPS), which are involved in the immune and inflammatory responses. Adrenocorticotropic hormone (ACTH) and angiotensin II, hormones that regulate the adrenal cortex, likewise regulate release of cytokines from adrenal glands. Dopamine inhibits aldosterone release from the adrenal cortex. Therefore, effects of dopamine on IL-6 and TNF release from rat adrenal zona glomerulosa were investigated. Primary cultures of rat adrenal zona glomerulosa cells were exposed to test agents and IL-6 and TNF release determined by the 7TD1 and WEHI bioassays, respectively. Dopamine increased basal IL-6 release and potentiated IL-6 release stimulated by ACTH, LPS or IL-1β. Dopamine inhibited basal and secretagogue-stimulated (LPS and IL-1β) TNF release. These effects of dopamine were mediated by D2 receptors because N-04 37, a D2 agonist, had effects on TNF and IL-6 release identical to those of dopamine. Therefore, dopamine, through D2 receptors, regulates the release of IL-6 and TNF from adrenal cells. Because TNF and IL-6 regulate adrenal steroid release, these cytokines may serve as autocrine or paracrine mediators of adrenal gland function. Allan M Judd, Department of Zoology, Brigham Young University, Provo, UT 84602, USA

Effect of composition of incubation medium on aldosterone and corticosterone production by isolated rat zona glomerulosa cells

1982 ◽  
Vol 94 (2) ◽  
pp. 211-224 ◽  
Author(s):  
D. J. Campbell
Keyword(s):  
Angiotensin Ii ◽  
Cyclic Amp ◽  
Incubation Medium ◽  
Zona Glomerulosa ◽  
Bicarbonate Buffer ◽  
Angiotensin Ii Antagonist ◽  
Aldosterone Production ◽  
Glomerulosa Cells ◽  
Zona Glomerulosa Cells

The role of the composition of the incubation medium in determining the steroidogenic responsiveness of collagenase-dispersed rat zona glomerulosa cells was examined by studying the effect on production of aldosterone and corticosterone of (1) changes in the bovine serum albumin (BSA) concentration in Krebs–Ringer bicarbonate buffer (KRBGA), (2) dialysis of the BSA and (3) comparison of KRBGA with 'modified' Medium 199. Medium 199 was modified so that its electrolytic content was identical to that of KRBGA. Compared with 0·1–0·2% BSA in KRBGA, BSA concentrations of 0·5 and 4% caused inhibition of both basal and K+-stimulated, but not angiotensin II-stimulated steroidogenesis. This inhibitory property of BSA was not removed by dialysis. The BSA did, however, contain a dialysable factor which increased both basal steroidogenesis and the steroidogenic response to maximal K+ and angiotensin II stimulation. Both incubation media contained 0·2% BSA for the comparison of KRBGA with modified Medium 199. Modified Medium 199 increased both basal steroidogenesis and the aldosterone response to K+ stimulation (per cent increase above basal) by two- to threefold compared with KRBGA, with smaller increases in the response to ACTH and 5-hydroxytryptamine (5-HT) and a decrease in the response to cyclic AMP. In contrast, modified Medium 199 increased the aldosterone response to angiotensin II by sevenfold, from 60% (in KRBGA) to 420%. In KRBGA, angiotensin II inhibited K+-stimulated aldosterone production. This effect was produced by concentrations of angiotensin II below the threshold for steroidogenesis and could be reproduced with the angiotensin II antagonist [Sar1, Ileu8]-angiotensin II. Angiotensin II did not inhibit K+-stimulated aldosterone production in modified Medium 199. These data emphasize the importance of the composition of the incubation medium in determining the steroidogenic responsiveness of rat zona glomerulosa cells in vitro. Furthermore, these data indicate that the steroidogenic response to angiotensin II, compared with K+, ACTH, 5-HT and cyclic AMP, is more readily influenced by other, as yet unidentified, factors in the incubation medium, and are consistent with recent evidence that angiotensin II and K+ do not share a common mode of action on steroidogenesis by these cells.

Effect of exposure to hypoxia from birth on aldosterone in rabbits: role of unesterified fatty acids

1997 ◽  
Vol 272 (4) ◽  
pp. R1084-R1087 ◽  
Author(s):  
H. Raff ◽  
B. M. Jankowski ◽  
T. L. Goodfriend ◽  
J. E. Baker ◽  
P. E. Papanek
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Zona Glomerulosa ◽  
Hypoxic Hypoxia ◽  
Adrenocortical Function ◽  
Total Serum ◽  
Aldosterone Production ◽  
Glomerulosa Cells ◽  
Zona Glomerulosa Cells

Hypoxia and fluid and electrolyte disturbances are serious risks to normal postnatal development. Because a decrease in inspired O2 (hypoxic hypoxia) inhibits aldosterone synthesis in the adult and aldosterone controls water and electrolyte balance, we studied adrenocortical function in rabbits exposed to normobaric normoxia or hypoxic hypoxia (fraction of inspired O2 0.09) from birth. At 21 days of age, rabbits were anesthetized, the adrenals were rapidly removed, and the adrenal capsules containing mostly zona glomerulosa cells were separated. Cells were dispersed with collagenase and studied in vitro. Hypoxia in vivo resulted in a 73% decrease in basal aldosterone release and a 86% decrease in adenosine 3',5'-cyclic monophosphate-stimulated aldosterone release in vitro. We hypothesized that increased unesterified fatty acids could be partly responsible for inhibition of aldosterone synthesis. Total serum unesterified fatty acids in hypoxic kits were significantly increased (298 +/- 14 micromol/l) compared with normoxic kits (184 +/- 31 micromol/l). When cells from hypoxic rabbits were washed with fatty acid-free albumin and studied under conditions devoid of fatty acids, aldosterone production was partially restored. Corticosterone production was not affected by washing. Washing had no effect on aldosterone synthesis by cells from normoxic rats. Finally, exposing washed zona glomerulosa cells to oleic acid (10-50 microM) inhibited aldosteronogenesis. We conclude that exposure to hypoxia from birth attenuates aldosterone production in part due to an increase in levels of unesterified fatty acid levels.

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