Catecholamine stimulation of cortisol secretion by 3-day primary cultures of purified zona fasciculata/reticularis cells isolated from bovine adrenal cortex

1988 ◽  
Vol 57 (1-2) ◽  
pp. 139-147 ◽  
Author(s):  
S.W. Walker ◽  
E.R.T. Lightly ◽  
S.W. Milner ◽  
B.C. Williams
Keyword(s):  
Adrenal Cortex ◽  
Primary Cultures ◽  
Zona Fasciculata ◽  
Cortisol Secretion ◽  
Stimulation Of

PROPERTIES OF RAT ADRENAL ZONA RETICULARIS CELLS: PRODUCTION AND STIMULATION OF CERTAIN STEROIDS

1979 ◽  
Vol 83 (3) ◽  
pp. 435-447 ◽  
Author(s):  
J. B. G. BELL ◽  
R. P. GOULD ◽  
P. J. HYATT ◽  
J. F. TAIT ◽  
S. A. S. TAIT
Keyword(s):  
Adrenal Cortex ◽  
Significant Role ◽  
Zona Fasciculata ◽  
Cell Type ◽  
Acth Stimulation ◽  
Specific Stimulus ◽  
Zona Reticularis ◽  
Cell Pool ◽  
Stimulation Of

The outputs of corticosterone, deoxycorticosterone and androstenedione from dispersed, purified rat adrenal zona reticularis and zona fasciculata cells have been measured by radioimmunoassay. Preferential production of deoxycorticosterone by zona reticularis cells was demonstrated by their higher basal deoxycorticosterone: corticosterone ratio when compared with zona fasciculata cells. Adrenocorticotrophin (ACTH) stimulated corticosterone output by all cell pools prepared by unit gravity (1 g) sedimentation, zona fasciculata cells being stimulated 130-fold compared with 20-fold for the zona reticularis cells in relation to their basal corticosterone output. In every cell pool, ACTH stimulated the output of corticosterone more than it stimulated the output of deoxycorticosterone. In parallel cell preparations, it was shown that ACTH increased the conversion of tracer amounts of radioactive deoxycorticosterone to corticosterone and decreased the conversion of radioactive corticosterone to 11-dehydrocorticosterone. Adrenocorticotrophin did not increase the conversion of radioactive deoxycorticosterone to total 11-oxygenated steroids (corticosterone+ 11-dehydrocorticosterone). It is unlikely therefore that ACTH stimulates 11 β-hydroxylation. Data indicate that the ratio of deoxycorticosterone to total 11-oxygenated steroids (corticosterone +11-dehydrocorticosterone) is characteristic for each cell type, and that this ratio will be relatively independent of ACTH stimulation or the amount of pregnenolone substrate available. Basal androstenedione outputs were similar for both types of cell, and ACTH stimulation was very small, being slightly greater for zona fasciculata than for zona reticularis cells. The contribution of the zona reticularis cells to the basal output of any steroid by the cells of the inner two zones of the adrenal cortex of the rat was relatively small (20% for deoxycorticosterone and 10% for corticosterone) and was even less after stimulation by ACTH. Unless a specific stimulus can be found, therefore, a significant role for the zona reticularis cannot yet be established.

Effect of adrenocorticotrophin on cortisol and androstenedione secretion from dispersed cells of guinea-pig adrenal zonae fasciculata and reticularis

1986 ◽  
Vol 109 (3) ◽  
pp. 399-404 ◽  
Author(s):  
W. R. Robertson ◽  
B. Davison ◽  
D. C. Anderson ◽  
J. Frost ◽  
A. Lambert
Keyword(s):  
Body Weight ◽  
Guinea Pig ◽  
Adrenal Cortex ◽  
Zona Glomerulosa ◽  
Adrenal Weight ◽  
Zona Fasciculata ◽  
Cortisol Secretion ◽  
Acth Stimulation ◽  
17 Hydroxyprogesterone ◽  
Dispersed Cells

ABSTRACT We have studied cortisol and androstenedione secretion by dispersed cells of the outer zona fasciculata (ZF) plus zona glomerulosa, and the inner zona reticularis (ZR) plus medulla of the guinea-pig adrenal. The ZF and ZR were microdissected apart, the cells dispersed and incubated (200 000 cells/ml) for 90 min in the presence of adrenocorticotrophin (ACTH; 500 ng/l), dibutyryl cyclic AMP (dbcAMP; 1 mmol/l), pregnenolone, 17-hydroxypregnenolone, 17-hydroxyprogesterone, 11-deoxycortisol and 21-deoxycortisol. The steroid concentrations were 5–25 μmol/l. Cortisol secretion was assayed by radioimmunoassay. There was no detectable cortisol secretion ( < 50 nmol/l) from the ZR in the controls (no additive) or after dbcAMP stimulation. Adrenocorticotrophin-stimulated cortisol secretion was also low (range < 50–340 nmol/l). In contrast the ZF secreted 177–379 (control), 828–2052 (dbcAMP) and 2863–9735 (ACTH) nmol cortisol/l. There was no detectable (i.e. < 2 nmol/l) cAMP production by ZR or ZF either basally (no ACTH) or after ACTH stimulation (500 ng/l). Challenge of the ZR cells with each cortisol precursor steroid (5 μmol/l) increased (P < 0·05) cortisol secretion over that seen with the corresponding basal and ACTH-stimulated controls. Thus pregnenolone, 17-hydroxypregnenolone, 17-hydroxyprogesterone, 11-deoxycortisol and 21-deoxycortisol (converted directly to cortisol by 21-hydroxylase) gave rise to (mean ± s.d., n = 4) 406 ± 86, 680 ± 180, 1307 ± 111, 1141 ± 234 and 3160 ± 419 nmol cortisol/l respectively. The corresponding figures for the ZF prepared from the same adrenal cortex were 3919 ± 309, 4122 ± 97, 5766 ± 615, 5035 ± 260 and 6954 ± 755 nmol cortisol/l. With pregnenolone (25 μmol/l), cortisol secretion increased to 7847 ± 1424 (ZR) and 12880 ± 982 nmol/l (ZF), a ZR:ZF ratio of 0·6 compared with 0·1 for pregnenolone at 5 μmol/l. Androstenedione was secreted in the basal state by both ZF and ZR in similar quantities, i.e. 3·7 ± 0·3 and 3·7 ± 0·4 nmol/l (n = 9 and 14 respectively). Both cell types were ACTH sensitive, with androstenedione secretion increasing to 28 ± 4·8 (ZF) and 12·5 ± 0·9 (ZR) nmol/l. There was a direct correlation (r = 0·924, P < 0·05) between total adrenal weight and body weight, between the percentage ZR in the cortex and adrenal weight (r = 0·96, P < 0·05) and between the percentage ZR and body weight (r = 0·981, P< 0·05). In a mature animal (800–1000 g) the ZR may occupy > 66% of the adrenal cortex. The mean diameters of cells from the ZF and ZR were 21 and 25 μm respectively. J. Endocr. (1986) 109, 399–404

scholarly journals 24-Dehydrocholesterol Reductase/Seladin-1: A Key Protein Differentially Involved in Adrenocorticotropin Effects Observed in Human and Rat Adrenal Cortex

Endocrinology ◽  
2009 ◽  
Vol 150 (9) ◽  
pp. 4180-4190 ◽  
Author(s):  
Marie-Claude Battista ◽  
Claude Roberge ◽  
Antoine Martinez ◽  
Nicole Gallo-Payet
Keyword(s):  
Oxidative Stress ◽  
Endoplasmic Reticulum ◽  
Adrenal Cortex ◽  
Intracellular Localization ◽  
Zona Fasciculata ◽  
Cortisol Secretion ◽  
Ros Production ◽  
Cytoplasmic Localization ◽  
Acth Treatment ◽  
And Oxidative Stress

Abstract DHCR24 (24-dehydrocholesterol reductase), or seladin-1, is one of the most expressed genes in the adrenal gland. Because the rat and human adult adrenal cortex differ in their respective functional properties, the aim of the present study was to verify whether seladin-1 may be differentially involved in basal and ACTH-stimulated steroidogenesis and oxidative stress management. Seladin-1 expression was predominantly observed in both human and rat zona fasciculata, with a predominant cytoplasmic localization in human cells and a nucleo-cytoplasmic distribution in rat cells. In human fasciculata cells, localization of the protein was primarily associated with the endoplasmic reticulum. Although its expression was increased by ACTH, its intracellular localization was not altered by ACTH treatment (10 nm) or by the seladin-1 inhibitor U18666A (75 nm). Preincubation with U18666A did not modify the ACTH-induced increase in cortisol secretion but abolished the ACTH-induced increase in dehydroepiandrosterone secretion. In rat fasciculata cells, ACTH induced a massive redistribution of seladin-1 from the cytoplasm (cis-Golgi apparatus) to the nucleus, which was inhibited by preincubation with U18666A. Preincubation with U18666A also decreased ACTH-induced seladin-1 and 11β-hydroxylase protein expression as well as corticosterone production, increased ACTH-induced ROS production but decreased ACTH-induced expression of the detoxifying protein aldo-ketoreductase 1b7. Thus, protection against acutely elevated ACTH-induced oxidative stress in rat fasciculata cells is correlated with nuclear relocalization of seladin-1 and its effects on cellular detoxifying machinery. Altogether, these results indicate that seladin-1 expression and intracellular localization are correlated with both the intensity and nature of ACTH-induced steroidogenesis and resultant oxidative stress.

Adrenergic and Cholinergic Regulation of Cortisol Secretion from the Zona Fasciculata/Reticularis of Bovine Adrenal Cortex

1991 ◽  
Vol 17 (1-2) ◽  
pp. 237-265 ◽  
Author(s):  
S. W. Walker ◽  
E. R. T. Lightly ◽  
C. Clyne ◽  
B. C. Williams ◽  
I. M. Bird
Keyword(s):  
Adrenal Cortex ◽  
Zona Fasciculata ◽  
Cortisol Secretion ◽  
Cholinergic Regulation

Dose-dependent effects of angiotensin II, acetylcholine and vasopressin on the cytosolic concentration of Ca2+ in suspension primary cultures of zona fasciculata/reticularis cells from bovine adrenal cortex

1991 ◽  
Vol 6 (2) ◽  
pp. 197-203 ◽  
Author(s):  
S. W. Walker ◽  
M. W. J. Strachan ◽  
M. Nicol ◽  
B. C. Williams ◽  
I. M. Bird
Keyword(s):  
Angiotensin Ii ◽  
Suspension Culture ◽  
Adrenal Cortex ◽  
Primary Cultures ◽  
Intracellular Concentration ◽  
Zona Fasciculata ◽  
Adrenocortical Cells ◽  
Intracellular Pool ◽  
Intracellular Ca ◽  
Dose Dependent

ABSTRACT The effects of angiotensin II (AII), acetylcholine and vasopressin on the intracellular concentration of Ca2+ have been little studied in adrenocortical cells from the zona fasciculata/reticularis (ZFR). Primary cultures of bovine ZFR cells maintained in suspension culture for 72 h produce cortisol in response to AII (0·1 μm), acetylcholine (0·1 mm) and vasopressin (1 μm). This response is accompanied by a breakdown of membrane phosphoinositides from [3H]inositol-prelabelled cells. Using cells loaded with the Ca2+ indicator fura-2, the intracellular concentration of Ca2+ was measured in response to increasing doses of all three agonists and found to increase in a graded fashion in each case. The basal intracellular concentration of Ca2+ was 75±3 nm (mean±s.e.m., n=52), rising to a maximum 1·82±0·14-fold (n=6) for AII (0·1 μm), 1·35±0·05-fold (n=7) for acetylcholine (0·1 mm) and 1·27±0·10-fold (n=6) for vasopressin (1 μm). In the case of AII and acetylcholine, agonists were added sequentially in medium of normal extracellular Ca2+ concentration (1·2 mm) or in medium in which the Ca2+ concentration was buffered to approximate to the intracellular concentration of Ca2+ (75–100 nm). Evidence was thereby obtained that both AII and acetylcholine mobilize a common intracellular pool of Ca2+. Our findings suggest that these three agonists, all of which stimulate phospholipase C, increase intracellular Ca2+ through a mechanism which depends, at least in part, on the release of Ca2+ from a common intracellular pool.

Respiratory Metabolism During Pregnancy in the Guinea Pig

1957 ◽  
Vol 190 (3) ◽  
pp. 425-428 ◽  
Author(s):  
Richard M. Hoar ◽  
William C. Young
Keyword(s):  
Heart Rate ◽  
Oxygen Consumption ◽  
Guinea Pig ◽  
Metabolic Rate ◽  
Adrenal Cortex ◽  
Guinea Pigs ◽  
Sharp Decrease ◽  
Zona Fasciculata ◽  
Respiratory Metabolism ◽  
Morphological Differences

Oxygen consumption and heart rate during pregnancy were measured in untreated, thyroxin-injected and thyroidectomized guinea pigs given I131. From impregnation until parturition, oxygen consumption increased 7.9% in untreated females. The increase continued until 5 days postpartum when a sharp decrease occurred. The increase is not accounted for by growth of the fetal mass. Comparable increases occurred in thyroxin-injected (16.2%) and thyroidectomized (11.9%) females, although the levels throughout were higher and lower, respectively, than in intact females. Heart rate did not increase. On the contrary, statistically significant decreases occurred in the untreated and thyroxin-injected females. Although the mechanism associated with the increased metabolic rate is not known, the possibility of thyroid participation would seem to be excluded. Involvement of the adrenal cortex is suggested by morphological differences in the cells of the zona fasciculata in pregnant and nonpregnant females and by evidence cited from other studies.

scholarly journals Coregulatory protein–orphan nuclear receptor interactions in the human adrenal cortex

2005 ◽  
Vol 186 (1) ◽  
pp. 33-42 ◽  
Author(s):  
Sinead N Kelly ◽  
T Joseph McKenna ◽  
Leonie S Young
Keyword(s):  
Transcriptional Regulation ◽  
Adrenal Cortex ◽  
Nuclear Receptor ◽  
Tumour Cell Line ◽  
Zona Fasciculata ◽  
Orphan Nuclear Receptor ◽  
Response Element ◽  
Coregulatory Proteins ◽  
Coregulatory Protein ◽  
In Cells

The capacity of the adrenal to produce steroids is controlled in part through the transcriptional regulation of steroid enzymes. The orphan nuclear receptor steroidogenic factor 1 (SF-1) is central to the transcriptional regulation of all steroid hydroxylase enzymes, whereas nur77 can preferentially regulate steroid enzyme genes relevant to cortisol production. We hypothesised that, in the presence of secretagogues, SF-1 and nur77 may differentially interact with coregulatory proteins in the human adrenal cortex. Both coregulatory proteins, steroid receptor coactivator (SRC-1) and silencing mediator for retinoid and thyroid hormones (SMRT), were found to be expressed in the zona fasciculata and reticularis in the human adrenal cortex, but were largely absent from the zona glomerulosa. Both coregulatory proteins were colocalised with SF-1 and nur77. In the H295R adrenal tumour cell line, SF-1 and nur77 transcripts were increased in cells in the presence of forskolin, whereas nur77 mRNA was also induced with angiotensin II (AII). The coactivator SRC-1 mRNA was increased in the presence of both forskolin and AII. Forskolin induced recruitment of SRC-1 to the SF-1 response element and induced SRC-1–SF-1 interactions, whereas AII increased recruitment of SRC-1 to the nur77 response element and induced SRC-1–nur77 interactions. The corepressor SMRT interacted with SF-1 in the presence of AII and with nur77 in cells treated with forskolin. Orphan nuclear receptor–coregulatory protein interactions may have consequences for the regulation of key steroidogenic enzymes in the human adrenal cortex.

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