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I M Bird

Publications and source records attributed to I M Bird.

At least 73 records · Page 4Linked to original sources
PubMed1993

Angiotensin increases aldosterone synthase mRNA levels in human NCI-H295 cells.

Understanding the regulation of aldosterone secretion has been hampered by the lack of a cell culture system that remains chronically responsive to angiotensin stimulation. NCI-H295 cells, cultured from a human adrenocortical tumor, express the three major pathways of adrenal steroidogenesis and produce small amounts of aldosterone during basal culture. We have determined changes in aldosterone production and in aldosterone synthase (AS, P45011B2) mRNA levels in these cells in response to angiotensin II (AII) and forskolin. Culture of NCI-H295 cells with 10(-7) M AII or with 10(-5) M forskolin stimulated aldosterone production and increased AS mRNA levels, though the effect of AII was greater. When cells were cultured with increasing concentrations of AII from 10(-11) through 10(-8) M, a dose-dependent increase in AS mRNA levels paralleled increases in aldosterone production. In view of these findings, these human adrenocortical cells should be useful for exploring mechanisms regulating aldosterone production.

Adrenal Cortex Neoplasms↗
PubMed1993

Effect of angiotensin II on human luteinized granulosa cells.

OBJECTIVE: To determine if human luteinized granulosa cells (GCs) exhibit angiotensin II (AII) receptors and if AII affects GC steroidogenesis. DESIGN: Luteinized GCs were either treated immediately or maintained in culture for up to 6 days before use. The presence of functional AII receptors was evaluated using the following criteria: [1] receptor binding of 125I-labeled AII; [2] changes in inositol phosphate production; [3] effects on progesterone (P) production; and [4] effects on cyclic adenosine 3':5' monophosphate (cAMP) production. RESULTS: No specific binding of 125I-labeled AII was observed using human luteinized GCs in culture. Angiotensin II did not alter the formation of [3H] inositol phosphates, nor did it alter the release of P from freshly isolated cells or cells that had been in culture for 5 days. Angiotensin II was also unable to alter basal or gonadotropin-stimulated cAMP production. These data suggest that human luteinized GCs exhibit little or no type 1 or type 2 AII receptors. CONCLUSION: This observation is in contrast to reports in rat granulosa that exhibit AII receptors. The reason for a lack of AII receptor expression on human luteinized GCs is unknown but could be because of luteal phase down regulation with gonadotropin-releasing hormone agonist or the subsequent stimulation with gonadotropins in these women before follicular stimulation.

Angiotensin II↗
PubMed1993

Angiotensin-II stimulates an increase in cAMP and expression of 17 alpha-hydroxylase cytochrome P450 in fetal bovine adrenocortical cells.

While known to be a potent activator of phosphoinositidase C, angiotensin II (A-II) also causes a small but significant increase in cAMP production through the type 1 A-II (AT1) receptor in bovine adrenocortical cells (Mol Cell Endocrinol 81:33-41, 1991). We have carried out studies on primary cultures of fetal bovine adrenocortical cells to examine the effects of A-II on the expression of cytochrome P450 17 alpha-hydroxylase (P450c17), which is known to be regulated in a cAMP-dependent fashion. Prolonged treatment (48 h) of cells with A-II (10(-7) M) did not give rise to a detectable increase in P450c17 as measured by immunoblotting, although both A-II and the protein kinase C activator, 12-O-tetradecanoylphorbol 13-acetate (TPA) attenuated the large increase in P450c17 induced by ACTH (10(-8) M). A-II alone (10(-7) M) however, caused a time-dependent increase in cAMP secretion, reaching 8-fold within 3 h. Prolonged treatment of cells with A-II also resulted in a 3-fold increase in P450c17 mRNA within 12 h (10(-7) M), and a dose-dependent increase in 17 alpha-hydroxylase activity within 48 h (16.4-fold max at 10(-7) M). The stimulatory actions of A-II alone (10(-7) M) on cAMP levels, P450c17 mRNA, and 17 alpha-hydroxylase activity were much smaller than in response to ACTH (10(-8) M), but were largely reproduced by TPA (10(-7) M), suggesting a role for protein kinase C in mediating these responses to A-II. These findings indirectly support the hypothesis that A-II alone can stimulate an increase in cAMP in adrenocortical cells. Such a stimulation of cAMP may then result in increased expression of steroidogenic enzymes, as we have shown is the case for P450c17 expression. However, A-II in the presence of ACTH appears to attenuate the ACTH-stimulated expression of P450c17.

Adrenal Cortex↗
PubMed1993

Human NCI-H295 adrenocortical carcinoma cells: a model for angiotensin-II-responsive aldosterone secretion.

Excessive secretion of aldosterone from the adrenal results in the most common form of endocrine hypertension. An understanding of the regulatory processes involved in aldosterone synthesis and release is needed to define the biomolecular mechanisms controlling excessive production of aldosterone. However, in vitro studies regarding the regulatory mechanisms of human aldosterone production have been limited because of difficulties in obtaining tissue and the subsequent isolation of aldosterone-secreting glomerulosa cells. Herein we describe an adrenocortical carcinoma cell line, NCI-H295, which provides a suitable angiotensin-II (AII)-responsive model system to investigate the acute and chronic regulation of aldosterone synthesis. The cells were characterized with regard to the effects of AII on second messenger systems, aldosterone release, and levels of aldosterone synthase (P450c18) mRNA. In the presence of lithium, AII caused a rapid, but transient, increase in the production of inositol tris- and bisphosphates, whereas a prolonged gradual accumulation of inositol monophosphate occurred. Treatment with AII resulted in a 4.5-fold increase in total inositol phosphates in a concentration-dependent manner and an increase in intracellular cytoplasmic free Ca2+. Significant increases in aldosterone (3.5-fold) were detected within 1 h of AII addition. Aldosterone release occurred in a concentration-and time-dependent manner. The type 1 AII (AT1) receptor was shown to be responsible for activation of phosphoinositidase-C, increased intracellular free Ca2+, and aldosterone production, as determined by use of the AT1 receptor antagonist DuP753. In addition, AII treatment resulted in a time-dependent increase in levels of P450c18 mRNA, as detected by RNAse protection assay. In summary, NCI-H295 cells provide a valuable model system to define mechanisms regulating human aldosterone production.

Adrenal Cortex Neoplasms↗
PubMed1993

Regulation of human adrenal carcinoma cell (NCI-H295) production of C19 steroids.

The regulation of biosynthesis of the adrenal C19 steroids (the so-called adrenal androgens) remains unclear. Understanding adrenal production of C19 steroids is important when the benefits of these steroids are considered on processes and diseases associated with aging. In vitro studies defining the mechanisms that regulate the production of human adrenal C19 steroids have been limited because of the difficulties in obtaining adrenal tissue. A cell line that retains differentiated adrenal functions would greatly facilitate research in this area. Herein, we describe the use of the human adrenocortical tumor H295 cell line as a model to evaluate mechanisms controlling C19 and C21 steroid production. The cells were characterized with regard to ACTH, forskolin, and dibutyryl cAMP (dbcAMP) responsiveness, as measured by increased cAMP production, synthesis of steroids, and induction of 17 alpha-hydroxylase cytochrome P450 (P450c17). Forskolin and dbcAMP, which were more effective than ACTH, enhanced the production of cortisol, dehydroepiandrosterone (DHEA), DHEA sulfate (DHEAS), and androstenedione over a 48-h treatment period. Comparison of the relative amounts of measured steroid secreted under forskolin treatment indicated that the primary product was cortisol (70%), followed by androstenedione (14%), DHEA (9%), and DHEAS (7%). Cortisol was also demonstrated to be the major steroid product by examination of UV-detectable steroids after high performance liquid chromatographic separation. The increases in steroid production caused by ACTH, forskolin, and dbcAMP occurred in a concentration- and time-dependent manner. A key enzyme in the production of C19 steroids is P450c17. ACTH, forskolin, and dbcAMP increased the activity of 17 alpha-hydroxylase by approximately 2.5-, 10-, and 10-fold, respectively. These effects on enzyme activity occurred in a concentration-dependent manner and coincided with increased levels of P450c17 mRNA. In summary, H295 cells should provide a much-needed model to study mechanisms controlling the secretion of glucocorticoids and C19 steroids, because steroid production in these cells is hormonally controlled and associated with the induction of P450c17.

Adrenal Cortex Hormones↗
PubMed1992

Studies of hormone-sensitive and -insensitive pools of phosphoinositides in cultured bovine zona fasciculata/reticularis cells. Evidence that acetylcholine and angiotensin II stimulate the breakdown of a common pool of phosphoinositides.

The effects of acetylcholine (ACh) and manganese pre-incubation on angiotensin II (AII)-stimulated incorporation of [3H]inositol into phosphoinositide, phosphoinositol and free inositol fractions of adrenocortical cells isolated from the bovine zona fasciculata/reticularis (zfr) were investigated. In cells pre-labelled for 6 hr with [3H]inositol, ACh and AII stimulated the incorporation of cytosolic [3H]inositol into a common hormone-sensitive pool of phosphoinositides, which was distinct from the non-hormone-sensitive pool labelled in the presence of manganese. Regression analysis of the cortisol versus [3H]inositol headgroup responses for both AII (10(-11)-10(-7) M) and ACh (10(-9)-10(-3) M) showed that the gradients of these responses were not significantly different. These data provide strong evidence that in cultured bovine zfr cells, ACh and AII stimulate the breakdown and resynthesis of a common pool of phosphoinositides.

Acetylcholine↗
PubMed1992

Angiotensin II receptors on human fetal adrenal cells.

OBJECTIVE: Our objective was to determine if angiotensin II receptors are present on adrenal cells isolated from the human fetal zone and neocortex and to investigate if angiotensin II affects steroid production by these cells. STUDY DESIGN: Primary cultures of both fetal zone and neocortex cells were prepared from fetal adrenal glands. Experiments were conducted to examine the binding of radiolabeled angiotensin II, angiotensin II activation of phospholipase C, and angiotensin II effects on steroidogenesis. RESULTS: The majority of angiotensin II binding sites were of the type 1 subtype, as determined by displacement of radiolabeled angiotensin with specific receptor antagonists. Angiotensin II caused an increase in tritiated inositol phosphate accumulation in both neocortex and fetal zone cells. This increase could be blocked by type 1 angiotensin II receptor antagonists. Angiotensin II stimulated the production of cortisol, dehydroepiandrosterone, and dehydroepiandrosterone sulfate production during treatment for 2 days. The stimulation by angiotensin II, however, was substantially less than that seen in response to corticotropin treatment. CONCLUSIONS: The human fetal adrenal gland contains type 1 angiotensin II receptors early in gestation. The number of these receptors, albeit low, is sufficient to activate inositol phosphate production and steroidogenesis.

Adrenal Glands↗
PubMed1992

Identification and metabolism of phosphoinositol species formed on angiotensin II stimulation of zona fasciculata-reticularis cells from the bovine adrenal cortex.

The identity of phosphoinositol isomers accumulating on stimulation of primary cultures of bovine adrenocortical zona fasciculata/reticularis cells with angiotensin II (AII), in the presence of Li+, has been established by chromatographic separation on a MonoQ HR5/5 column. The metabolism of inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) in a broken cell preparation has also been studied in the absence or presence of added ATP. Our results show that Ins(1,4,5)P3 is formed within 5 s of stimulation of whole cells, but is rapidly converted to Ins(1,3,4)P3 through an Ins(1,3,4,5)P4 intermediate. All the phosphoinositol products accumulating on prolonged (15 min) stimulation of whole cells (Ins1P, Ins4P, Ins(1,3)P2, Ins(1,4)P2, Ins(1,3,4)P3, Ins(1,4,5)P3, Ins(1,3,4,5)P4) can be accounted for by the metabolism of Ins(1,4,5)P3 in broken cells, either through direct dephosphorylation in the absence of added ATP (Ins(1,4)P2, Ins4P) or through dephosphorylation of Ins(1,3,4,5)P4 formed in the presence of added ATP (Ins(1,3,4)P3, Ins(1,3)P2 and Ins1P). Our results provide further evidence to suggest that AII stimulates the rapid and sustained breakdown of phosphatidylinositol 4,5-diphosphate (PtdIns(4,5)P2) to form Ins(1,4,5)P3.

Angiotensin II↗
PubMed1992

Angiotensin-II acts via the type 1 receptor to inhibit 17 alpha-hydroxylase cytochrome P450 expression in ovine adrenocortical cells.

In this study we have investigated the effect of angiotensin-II (A-II) on cortisol production and 17 alpha-hydroxylase cytochrome P450 (P450(17 alpha)) expression in primary cultures of ovine adrenocortical cells and the A-II receptor subtypes that mediate these responses. While A-II alone had no stimulatory effect on cortisol secretion, it inhibited the cortisol response to ACTH (10(-8) M) in a dose-dependent manner (Ki, less than 0.1 nM; maximum inhibition, 60-80%). While prolonged treatment with ACTH (10(-8) M) increased the expression of P450(17 alpha), cotreatment with A-II (10(-8) M) also inhibited ACTH-stimulated expression, as determined by changes in mRNA, immunoreactive P450(17 alpha), and 17 alpha-hydroxylase activity. A study of the effects of the AT1 and AT2 receptor antagonists, DuP 753 and PD 123319, on binding of [125I]A-II to ovine adrenocortical cells showed that the A-II receptor population was predominantly of the AT1 subtype. The effects of A-II on inhibition of cortisol secretion in response to ACTH and the activation of phosphoinositidase-C in response to A-II alone were both fully antagonized by DuP 753, but not by PD 123319. Furthermore, the inhibitory effects of A-II on expression of P450(17 alpha), as measured at the levels of mRNA, immunoreactive protein, and enzyme activity, were reversed by DuP 753 (10(-5) M), but not PD 123319 (10(-5) M). We conclude that A-II has a potentially important role in the control of cortisol secretion and long term maintenance of P450(17 alpha) expression in the ovine adrenal cortex, and that the effects of A-II on both cortisol secretion and P450(17 alpha) expression are mediated through the AT1 receptor, which is coupled to phosphoinositidase-C.

Adrenal Cortex↗
PubMed1992

Interleukin-1 beta stimulates growth of adrenocortical cells in primary culture.

Using tritiated-thymidine incorporation as a measure of cell growth, interleukin-1 beta stimulated the growth of bovine zona fasciculata/reticularis adrenocortical cells after 72 h in primary culture. Within the range of 10-1000 pg/ml, interleukin-1 beta produced over 40% of angiotensin II-stimulated [3H]thymidine incorporation (P less than 0.005 compared with basal for 10 pg/ml and 1000 pg/ml; P less than 0.05 for 100 pg/ml; two-tailed unpaired Student's t-test). Interleukin-1 beta did not directly stimulate cortisol secretion. By stimulating adrenocortical growth, the increase in interleukin-1 during fever provides a potential mechanism for chronically raising glucocorticoid output. This study is the first demonstration of a long-term effect involving interleukin-1 beta on the adrenal cortex.

Animals↗
PubMed1992

Evidence for two distinct hormone-sensitive [3H]phosphoinositide pools in bovine adrenocortical zona fasciculata/reticularis cells stimulated with angiotensin II.

Bovine adrenocortical cells from the zona fasciculata/reticularis were isolated and their phosphoinositides labelled to a steady state with [3H]inositol in primary culture. Experiments performed on these cells in the presence of Li+ have shown that, over a period of 60 min, angiotensin II (AII; 10(-7) M) stimulated a linear increase in [3H]inositol phosphates that was sustained through the utilization of two hormone-sensitive subpools of prelabelled lipid (30% and 45% respectively), and a rapid resynthesis of [3H]phosphoinositide into one of these pools using cytosolic [3H]inositol. The 30% pool was used immediately on stimulation, and was sustained at a steady-state size of 10-15% during the first 30 min of stimulation through rapid resynthesis using cytosolic [3H]inositol. Only after 30 min, when the cytosolic [3H]inositol was depleted and resynthesis could no longer occur, did the additional 45% pool start to supply further substrate to the phospholipase C, thereby further sustaining the generation of [3H]inositol phosphates. Once this pool was depleted however (by approximately 60 min), [3H]inositol phosphate generation finally ceased. These findings establish the differential use of two metabolically distinct hormone-sensitive pools of phosphoinositide following AII stimulation in bovine adrenocortical cells, events which are dependent upon the availability of cytosolic inositol for phosphoinositide resynthesis.

Angiotensin II↗
PubMed1992

Further characterization of the steroidogenic responsiveness of purified zona fasciculata/reticularis cells from bovine adrenal cortex before and after primary culture: changing responsiveness to phosphoinositidase C agonists.

When bovine adrenocortical cells from the zona fasciculata/reticularis (zfr) are maintained in primary culture, cortisol secretion in response to acute stimulation with ACTH and adrenaline (which activate adenylate cyclase) is seen to increase steadily over the first 48 h, while secretion in response to angiotensin II and acetylcholine (which activate phosphoinositidase C) shows an initial decline in the first 24 h and a recovery to maximum after 48 h. We have investigated whether these discrepant changes in cortisol secretory response to the different agonists are due to changes in formation of the associated second messengers (cAMP or inositol phosphates), or altered coupling of these second messenger signals to steroid secretion. Increases in steroid secretion in response to ACTH and adrenaline were paralleled by increased cAMP. Steroid secretion in response to exogenous 8-bromoadenosine 3':5'-cyclic monophosphate also increased steadily during this 48-h period. Thus increased responsiveness was due to both increased second messenger formation and increased coupling to the steroid secretory response. The decreased steroid secretory response to angiotensin and acetylcholine after 24 h, and subsequent recovery after 48 h in culture, were accompanied by an increased formation of phosphoinositols after 24 h and a further increase by 48 h. However, the steroid secretory response to a combination of calcium ionophore and the protein kinase C activator, phorbol 12-myristate 13-acetate, was reduced after 24 h and recovered by 48 h of culture. Fura-2-loaded cells also showed an increase in intracellular [Ca2+] after 24 h in culture.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Adrenergic and cholinergic regulation of cortisol secretion from the zona fasciculata/reticularis of bovine adrenal cortex.

Inner zone cells, isolated from bovine adrenal cortex, secrete cortisol in response to both adrenergic and cholinergic agonists. The response to adrenaline (and other catecholamines) appears during culture, is evident by 24 h and reaches a maximum by 48-72 h, but is absent in freshly isolated cells. Pre-incubation of cultured cells with adrenaline leads to homologous desensitisation; the possibility that this may explain the absent response in freshly isolated cells is discussed. Cells show a dose-dependent cyclic AMP response but no increased membrane phosphoinositide turnover. In agreement, cortisol secretion is blocked by beta-receptor, but not alpha-receptor, antagonists. Schild analysis established that the response occurs through binding to a beta 1-receptor subtype, consistent with adrenergic innervation as opposed to an effect of circulating catecholamines. In contrast, cortisol secretion to AcCh was present in both freshly isolated cells and those in culture, reaching a maximum by 48-72 h in culture. The response was specifically blocked by muscarinic, but not nicotinic, antagonists. No effect on cyclic AMP formation was observed, but dose-dependent stimulation of phosphoinositide turnover occurred. HPLC analysis of the time-course of appearance of 3H-inositol labelled head groups (from cells pre-labelled with 3H-inositol) confirmed that AcCh activates a phosphoinositidase C. Intracellular Ca2+ oscillations were also measured from fura-2 loaded single cells in response to AcCh. Together with other pharmacological studies, these observations establish that AcCh acts through a M3 muscarinic receptor subtype in these cells. The possible significance of these findings in vivo is discussed.

Acetylcholine↗
PubMed1991

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.

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 cultured for 72 h produce cortisol in response to AII (0.1 microM), acetylcholine (0.1 mM) and vasopressin (1 microM). 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 microM), 1.35 +/- 0.05-fold (n = 7) for acetylcholine (0.1 mM) and 1.27 +/- 0.10-fold (n = 6) for vasopressin (1 microM). 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.

Acetylcholine↗
PubMed1990

Acetylcholine stimulates cortisol secretion through the M3 muscarinic receptor linked to a polyphosphoinositide-specific phospholipase C in bovine adrenal fasciculata/reticularis cells.

Zona fasciculata/reticularis (ZFR) cells, isolated from the bovine adrenal cortex, secreted cortisol in response to acetylcholine (AcCh). The response was present in freshly isolated cells and in cells maintained in primary culture, reaching a maximum after 48-72 h and thereafter declining. Cells maintained in primary culture for 72 h secreted cortisol with an ED50 at 1.2 x 10(-6) M. The potent inhibition of AcCh-stimulated secretion by atropine, and the relative ineffectiveness of nicotine or nicotinic antagonists, were consistent with a predominantly muscarinic response to AcCh in these cells. A selective M1-receptor agonist, McN-A-343, had no effect on cortisol secretion whereas the M3 antagonist, hexahydro-sila-difenidol, produced a dose-dependent inhibition of AcCh-stimulated cortisol secretion. These findings are consistent with AcCh mediating its effects on cortisol secretion through an M3 receptor. While AcCh had no effect on cAMP formation, a dose-dependent increase in [3H]phosphoinositols (identified using high-performance liquid chromatography (HPLC)) occurred in a manner that was not dependent on an influx of extracellular Ca2+. Detailed HPLC analysis of the formation of 3H-labelled phosphoinositols and glycerophosphoinositols from pre-labelled cells over the period 0-15 min showed that the earliest significant rise was in Ins(1,4,5)P3 at 5 s, followed by later rises in InsP1, InsP2 and Ins(1,3,4)P3. Additional studies using cells loaded with fura-2 indicator revealed a 1.6-fold increase in [Ca2+]i from a mean resting value of 75 nM in response to 10(-4) M AcCh. Furthermore, the rise in Ca2+ was not abolished by lowering extracellular Ca2+ to resting cytosolic levels, suggesting the mobilisation of an intracellular pool. These observations indicate that AcCh promotes rapid activation of a Ca2(+)-independent and polyphosphoinositide-specific phospholipase C, and that the Ins(1,4,5)P3 formed releases Ca2+ from an intracellular pool. The stimulation by AcCh of this signal transduction mechanism is consistent with our conclusion, based on the effects of the selective muscarinic agonist and antagonist on cortisol secretion, that the AcCh receptor is of the M3 subtype. We conclude that AcCh, acting through an M3 receptor coupled to phospholipase C, regulates cortisol secretion at the cellular level in bovine adrenal ZFR cells.

Acetylcholine↗
PubMed1990

Subclassification of beta-adrenoceptors responsible for steroidogenesis in primary cultures of bovine adrenocortical zona fasciculata/reticularis cells.

1. Forty eight hour primary cultures of purified bovine adrenocortical zona fasciculata/reticularis cells secreted hydrocortisone in response to stimulation with beta-adrenoceptor agonists. The observed order of potency was isoprenaline greater than noradrenaline greater than dobutamine greater than salbutamol greater than BRL37344. 2. Salbutamol acted as a partial agonist on these cells hence suggesting the presence of a beta 1-adrenoceptor. 3. Schild analysis of the hydrocortisone response to isoprenaline showed that the selective beta 1-antagonist practolol and the selective beta 2-antagonist ICI118,551 gave pA2 values of 6.85 and 7.17, respectively. These values were in close agreement with corresponding pA2 values previously obtained for the beta 1-adrenoceptor. 4. We conclude that beta 1-adrenoceptors are responsible for mediating catecholamine-stimulated hydrocortisone secretion from primary cultures of bovine zona fasciculata/reticularis cells.

Adrenergic beta-Agonists↗
PubMed1990

Vasopressin stimulates cortisol secretion and phosphoinositide catabolism in cultured bovine adrenal fasciculata/reticularis cells.

Cells isolated from the zona fasciculata/reticularis (ZFR) of the bovine adrenal cortex and maintained in culture were found to secrete cortisol in response to vasopressin stimulation. The increased cortisol secretion was dose dependent, with a threshold response at 1 nM and a maximal response (1.68-fold over basal) at 0.1 microM. In cells cultured in the presence of [3H]inositol (to prelabel the membrane phosphoinositide pool), stimulation with vasopressin in the presence of LiCl (10 mM) resulted in a similar dose-dependent increase in labelling of the phosphoinositol fraction, with a maximal response (1.45-fold over basal) at 10 nM. The increased labelling of the phosphoinositol fraction was independent of extracellular Ca2+ as it was not abolished in medium with [Ca2+] buffered to intracellular resting levels. This suggests that vasopressin stimulation results in the activation of a phosphoinositidase C. It is probable that cortisol secretion by bovine ZFR cells in response to vasopressin is dependent upon activation of this Ca2(+)-independent phosphoinositidase C. However, the small magnitude of the cortisol secretory response makes it unlikely that vasopressin is a primary regulator of cortisol secretion in vivo.

Acetylcholine↗