Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

---

Query: EC:4.6.1.1 (adenylate cyclase)
19,190 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

In order to investigate the expression and the regulation of steroidogenesis, human Leydig cells were fused with a functional mouse adrenal cell line (Y1). Six independent hybrid clones were analysed for hormone receptors and for cAMP and steroid response to ACTH, hCG, 8Br-cAMP or forskolin. All hybrids had lost hCG receptors and their ability to produce testosterone. With respect to the response of adenylate cyclase to ACTH and/or forskolin, hybrids could be classed into two groups. In the first group, the pattern of response was qualitatively similar to Y1 parental cells; The second group was far less responsive to ACTH than are Y1 cells, and when added together, forskolin and ACTH only had an additive effect. All hybrids responded to ACTH and 8Br-cAMP with an increased production of pregnenolone (P5). The amounts of P5 produced both under basal conditions and following 8Br-cAMP stimulation were significantly higher in three hybrids when compared to Y1 cells. However, the ability of two of these three hybrids to produce 20 alpha-dihydroprogesterone (20 alpha OHP4) was very low. The metabolism of [14C]P5 revealed that in one of these hybrids, there was a loss of 3 beta-hydroxysteroid dehydrogenase/isomerase whereas in the other case, there was a low 20 alpha-hydroxylase activity. The inhibition of cell growth by ACTH was related to the ability of the hormone to stimulate cAMP. Conversely, the inhibitory growth effects of 8Br-cAMP were not always inversely correlated with the ability of this nucleotide to stimulate P5 production. Since hybrids contained two mouse genomes and retained variable human chromosomes, these results suggest that extinction or enhancement of murine genes coding for some of the enzymes involved in steroidogenic response to ACTH was due to the regulation by human genes.
...
PMID:Inhibitory or stimulatory effect of human genes on the expression of adrenal function in human Leydig X Y1 cell hybrids. 298 67

The influence of follicular maturation on progesterone production by collagenase-dispersed hen granulosa cells was measured in short-term incubations. Granulosa cells of the largest follicle (F1) produced up to ten times more progesterone than cells from smaller follicles (F3-F5), not only in response to luteinizing hormone (LH), but also when stimulated by exogenous cyclic AMP or forskolin, both of which raise intracellular cyclic AMP levels by nonreceptor-mediated mechanisms. Moreover, when granulosa cell progesterone synthesis was stimulated by incorporating 25-hydroxy-cholesterol into the incubation medium, an identical pattern was obtained. This could be attributed to a corresponding increase in the specific activity of the mitochondrial cholesterol side-chain cleavage enzyme (20,22 desmolase). An increase in the apparent Vmax was observed without a change in the apparent Km values. Pregnenolone substrate at concentrations which raised progesterone production to levels similar to those observed in response to LH stimulation was utilized equally by granulosa cells of mature and developing follicles. However, at high pregnenolone concentrations, granulosa cells of mature follicles converted significantly more of the precursor to progesterone. Assay of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) showed that the enzyme has two Kms: a low Km present in cells of both mature and developing follicles, and a high Km found only in granulosa cells of more mature follicles. It is concluded that LH-promoted progesterone synthesis in granulosa cells of developing chicken follicles is restricted not so much by the availability of receptors and the competence of the adenylate cyclase/cyclic AMP system, but by the activity of key enzymes, principally the cholesterol-20,22 desmolase.
...
PMID:Influence of follicular maturation on luteinizing hormone-, cyclic 3',5'-adenosine monophosphate-, forskolin- and cholesterol-stimulated progesterone production in hen granulosa cells. 298 34

This study examines the activity of the adenylate cyclase system and that of some enzymes of the steroidogenic pathway of adrenal cells from 62-63 day old ovine fetuses. Synthetic corticotropin (ACTH1-24), cholera toxin and forskolin stimulated both cAMP and corticoid productions by freshly isolated adrenal cells. The cAMP response to ACTH1-24 was lower than that to forskolin. However, forskolin-induced steroidogenesis was significantly lower than the ACTH1-24-induced steroid output. Freshly isolated cells metabolized quickly [14C]-labeled pregnenolone mainly through the 17-deoxy pathway. The amounts of cortisol and of corticosterone formed, in the presence of exogenous pregnenolone, were roughly 15-fold higher than under maximal stimulation by ACTH1-24. When the cells were cultured for 6 days in the absence or presence of ACTH1-24 (10(-8) M) or forskolin (10(-5) M), a small development of the cAMP response to these factors was observed in the course of the experiment. However, the mechanism of this development appeared different, according to the conditions of culture. The amounts of corticosterone secreted on day 6 by ACTH1-24- or forskolin-treated cells were 2- to 4-fold higher than on day 1, whereas cortisol outputs were much lower on day 6 than on day 1. The response to ACTH1-24 of cells maintained in ACTH-free media decreased dramatically during the culture in terms of both cortisol and of corticosterone. On day 6 of the experiment, the metabolism of [14C]pregnenolone was lower than on day 1 under all 3 conditions of culture. Only the 3 beta-hydroxysteroid dehydrogenase/isomerase activity could be maintained by continuous treatment with forskolin. However, both ACTH1-24 and forskolin enhanced the production of pregnenolone from an endogenous substrate. In conclusion, these results present evidence that: 1) the adenylate cyclase system is not a bottleneck in the steroidogenic response to ACTH1-24 of freshly isolated adrenal cells from 62-63 day old ovine fetuses; 2) the main rate-limiting step for steroidogenesis by these cells is the availability of pregnenolone; 3) neither ACTH1-24 nor forskolin is able to maintain the activity of most enzymes involved in the metabolization of pregnenolone by cultured cells while increasing pregnenolone availability; 4) some inhibiting factors are involved in the loss of adrenal cells responsiveness to ACTH between days 50 and 100 of gestation, and they probably act mainly on the adenylate cyclase system.
...
PMID:Adrenal adenylate cyclase and steroidogenic activities of 63 day old ovine fetuses: in vitro effects of ACTH1-24 and forskolin. 312 Jul 97

Previously, we showed that 12-O-tetradecanoyl phorbol 13-acetate (TPA), an activator of protein kinase C, is a mitogen for fetal human definitive zone adrenocortical cells in culture. In the present experiments, TPA inhibited forskolin-stimulated cortisol, dehydroepiandrosterone (DHEA), and dehydroepiandrosterone sulfate (DHEAS) synthesis, and enhanced forskolin-stimulated progesterone and corticosterone synthesis. These changes in the pattern of steroidogenesis were shown to result from changes in enzyme activities after forskolin treatment. TPA increased forskolin-stimulated 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) 2-fold, while depressing forskolin-stimulated 17 c-hydroxylase to basal values DHEA sulfotransferase increased 3-fold on transfer of human adrenocortical cells from serum-containing to defined, serum-free medium; TPA inhibited this increase. Experiments in which TPA was added at various times during the time course of forskolin stimulation of 17 alpha-hydroxylase showed that TPA prevents the increase in the level of 17 alpha-hydroxylase, and does not have a direct inhibitory effect on 17 alpha-hydroxylase activity. TPA also inhibited stimulation of 17 alpha-hydroxylase by cAMP analogs, indicating that the inhibition of 17 alpha-hydroxylase by TPA is not due to an effect on adenylate cyclase. Previous experiments have shown that stimulation of intracellular cAMP is sufficient for androgen synthesis by the human adrenocortical cell, under defined, serum-free conditions, and that its high rate of androgen synthesis likely results from the relative levels of 3 beta-HSD, 17 alpha-hydroxylase, and DHEA sulfotransferase in the cell. Enzyme induction by cAMP results in increased production of both androgens and glucocorticoids, whereas activation of protein kinase C changes the balance of enzymes, resulting in increased non-17 alpha-hydroxylated steroid synthesis and decreased androgen and cortisol biosynthesis.
...
PMID:TPA inhibits the synthesis of androgens and cortisol and enhances the synthesis non-17 alpha-hydroxylated steroids in cultured human adrenocortical cells. 349 11

The steroidogenic responsiveness to ACTH of the fetal adrenal gland of several species increases strikingly during the last weeks of gestation. Using the ovine fetus model it has been shown that this spontaneous development involves modifications at different steps in the mechanism by which ACTH stimulates the cells: one located at the cell membrane, another beyond cAMP formation, namely, in the steroidogenic pathway. The first phenomenon is related to at least three processes: an increased number of ACTH receptors, an increased activity of adenylate cyclase, and an enhanced coupling of these entities. The enhanced steroidogenic capacity is related mainly to an increased ability to produce pregnenolone and to an enhanced activity in both 3 beta-hydroxysteroid dehydrogenase and 17 alpha-hydroxylase. Infusion of 115-120-day-old fetuses with ACTH for only 5 days induced development of most of these biochemical steps, indicating that ACTH is an important regulating hormone for the maturation of fetal adrenocortical function. However, when ovine fetal adrenal cells were cultured in the absence of ACTH, there was a spontaneous development of the hormone-sensitive adenylate cyclase system and of the steroidogenic pathway. Addition of ACTH to the culture medium accelerated and further enhanced these maturation processes. These results suggest that the in vivo maturation of ovine fetal adrenal is blocked by the presence in the fetal circulation of some inhibiting factors which are likely to be of extrapituitary origin. Therefore, the 'maturation' of ovine fetal adrenocortical function appears to be regulated in a coordinate fashion by stimulatory, ACTH and unknown inhibitory factors.
...
PMID:Ontogeny of the ACTH receptor, adenylate cyclase and steroidogenesis in adrenal. 609 87

The nature of the luteolysin in humans is unknown. Hydrogen peroxide (H2O2), notably released by activated leukocytes, is generated in the rat corpus luteum at luteolysis and evokes luteolytic-like effects in rat luteal cells. We, therefore, evaluated the actions of H2O2 in human luteinized granulosa cells. After 2 days of preculture with low levels of hCG, human granulosa luteal cells were placed in suspension culture for 1 h in the presence of isobutylmethylxanthine (100 microM). A 60-min challenge with hCG evoked dose-dependent stimulation of cAMP and progesterone production. H2O2 dose-dependently inhibited progesterone production (ED50, 50-100 microM) in the absence or presence of hCG and blocked hCG-stimulated cAMP accumulation. Inhibition of progesterone synthesis by H2O2 was near maximal within 5 min, whereas inhibition of cAMP accumulation was not evident until 60 min. Cell viability was unaffected by H2O2, and inhibition of cAMP was reversible, but inhibition of steroidogenesis was long-lasting. Progesterone production stimulated by 8-bromo-cAMP, 22-hydroxycholesterol, and pregnenolone was inhibited by H2O2 as was androstenedione-dependent estradiol production. These findings indicate that H2O2 blocked progesterone synthesis by inhibition of cholesterol side-chain cleavage cytochrome P450, 3 beta-hydroxysteroid dehydrogenase, aromatase, and/or 17 beta-hydroxysteroid dehydrogenase. While H2O2 blocked stimulation of cAMP accumulation in response to hCG and cholera toxin, this same response produced by forskolin or aluminum fluoride was unaffected by H2O2. Thus, H2O2 appears to uncouple LH (hCG) receptors by interruption of G-protein-dependent activation of adenylate cyclase. In summary, H2O2 evokes effects in isolated human granulosa luteal cells that are associated with luteal regression, which raises the interesting possibility that H2O2 may serve a role as a mediator of this process like that in the rat.
...
PMID:Hydrogen peroxide evokes antisteroidogenic and antigonadotropic actions in human granulosa luteal cells. 767 98

We have developed and characterized a primary cell culture system to study the regulation of 17 alpha-hydroxylase cytochrome P450 (P45017 alpha) gene expression in bovine thecal cells. Conditions have been established for the dispersal and growth of thecal cells isolated from bovine follicles, which maintain the expression of P45017 alpha for up to 8 days. Bovine theca interna cells were grown to subconfluence and transferred into medium containing forskolin, a stimulator of adenylate cyclase. Levels of P45017 alpha transcripts reached a maximum value after 48 h of stimulation with forskolin. Added progesterone was converted to 17 alpha-hydroxyprogesterone at a rate of 214 pmol/mg protein.h in cells treated with forskolin for 72 h, whereas in control cells, the rate was 9.2 pmol/mg protein.h after 72 h. This was reflected in a 10-fold increase in endogenous androstenedione production by forskolin-stimulated cells. Studies employing various growth factors suggest that transforming growth factor-beta, but not basic fibroblast growth factor, is a potent inhibitor of forskolin-induced 17 alpha-hydroxylase activity and androstenedione production in these cells. We have also characterized this cell culture system with respect to expression of other steroidogenic enzymes. Cholesterol side-chain cleavage cytochrome P450 and 3 beta-hydroxysteroid dehydrogenase transcripts as well as endogenous progesterone accumulation were increased in response to forskolin stimulation. On the other hand, aromatase cytochrome P450 expression was undetectable. The ability to maintain bovine thecal cells, which retain 17 alpha-hydroxylase activity, in culture will provide a model system to study the regulation of expression of the P45017 alpha gene in the bovine ovary.
...
PMID:Maintenance and regulation of 17 alpha-hydroxylase expression by bovine thecal cells in primary culture. 767 80

Seminiferous tubules prepared from adult rats cultured for 48 h in serum-free conditions produce multiple biological factors that modulate Leydig cell steroidogenic function in vitro. Using gel filtration chromatography, it was shown that seminiferous tubular culture medium (STCM) contained at least three inhibitory activities designated AI, AII, and AIII that inhibited testosterone production by purified Leydig cells. The factor that induced AIII activity, designated Leydig cell inhibitor (LCI), was further purified to apparent homogeneity by sequential HPLC using gel permeation, C8-, C18-, C2/C18-reversed-phase, and microbore anion exchange columns. When this batch of purified factor was resolved by SDS-PAGE under reducing conditions, only a single silver stained band with an apparent M(r) of 21,000 was detected. Protein sequence analysis using about 100 pmol of purified LCI revealed that its N-terminus was blocked. Incubation of this highly purified factor with Percoll gradient purified Leydig cells induced a dose-dependent inhibition of hCG-stimulated testosterone production. LCI inhibited the basal testosterone production and hCG-stimulated cAMP production by Leydig cell dose-dependently. It also inhibited the forskolin- and cholera toxin-stimulated testosterone and cAMP production but had no apparent effect on the binding of 125I-labeled hCG to LH receptors. These data suggest that this LCI exerts its inhibitory action at steps beyond the LH receptors but prior to the cAMP formation by affecting the adenylate cyclase activity directly or indirectly through inhibition of the stimulatory G-protein (Gs-protein); however, it is also possible that it decreases the coupling of the receptors to the Gs-protein. LCI also inhibited the conversion of exogenously added 22R-hydroxycholesterol, pregnenolone, progesterone, and 17 alpha-hydroxyprogesterone to testosterone. However, it had no effect on the conversion of dehydroepiandrostenedione and androstenedione to testosterone. These data strongly suggest that LCI affects the steroidogenic enzymes metabolizing cholesterol to testosterone, the cytochrome P-450 side-chain cleavage (P-450SCC), and cytochrome P-450 17 alpha-hydroxylase/17,20-lyase (P-450C17). However, it has no effect on the 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) and 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) enzyme activities. Based on the results of the present study, it is apparent that this LCI is distinct from other known potent Leydig cells inhibitors such as interleukin-1 (IL-1) and transforming growth factor-beta (TGF-beta). The LCI appears to involve in the paracrine regulation of Leydig cell function.
...
PMID:Rat seminiferous tubular culture medium contains a biological factor that inhibits Leydig cell steroidogenesis: its purification and mechanism of action. 798 48

Intracellular effector systems which utilize PKA and PKC can be pharmacologically activated by forskolin and phorbol 12-myristate 13-acetate (PMA) and appear to be important for regulation of steroidogenesis by cells of the corpus luteum. In this study the effect of pharmacologic activation of PKA (forskolin) or PKC (PMA) on the activity of adenylate cyclase, cholesterol esterase, P450 cholesterol side chain cleavage (P450scc) and 3 beta-hydroxysteroid dehydrogenase/delta 5, delta 4 isomerase (3 beta HSD) was determined. Basal adenylate cyclase activity (as measured by intracellular and secreted cAMP) was extremely low in both large and small luteal cells. Forskolin stimulated adenylate cyclase activity in both large and small luteal cells but progesterone production was increased only in small cells. PMA inhibited progesterone production by large and forskolin-stimulated small cells without altering adenylate cyclase activity. Basal cholesterol esterase activity was greater in small than in large cells and was stimulated by forskolin only in small cells. PMA did not significantly alter cholesterol esterase activity in either cell type. Activity of P450scc or 3 beta HSD was measured by conversion of hydroxylated cholesterol derivatives (P450scc) or pregnenolone (3 beta HSD) to progesterone. Although basal progesterone production was 47 times greater in large than small cells, there was only 5.1 (P450scc) and 6.4 (3 beta HSD) times greater enzyme activity in large than in small luteal cells. Activation of PKA and/or PKC did not alter the activity of P450scc or 3 beta HSD in either cell type.(ABSTRACT TRUNCATED AT 250 WORDS)
...
PMID:Steroidogenic enzyme activity after acute activation of protein kinase (PK) A and PKC in ovine small and large luteal cells. 814 91

The appropriate expression of 3 beta-hydroxysteroid dehydrogenase/delta 5-->4-isomerase (3 beta-HSD) is vital for mammalian reproduction, fetal growth and life maintenance. Several isoforms of 3 beta-HSD, the products of separate genes, have been identified in various species including man. Current investigations are targeted toward defining the processes that regulate the levels of specific isoforms in various steroidogenic tissues of man. High levels of expression of 3 beta-HSD were observed in placental tissues. It has been generally considered that the multinucleated syncytiotrophoblastic cells are the principal sites of 3 beta-HSD expression and, moreover, that 3 beta-HSD expression is intimately associated with cyclic AMP-promoted formation of syncytia. Herein we report the presence of 3 beta-HSD immunoreactive and mRNA species in uninucleate cytotrophoblasts in the chorion laeve, similar to that in syncytia but not cytotrophoblast placenta. In vitro, 3 beta-HSD levels in chorion laeve cytotrophoblasts were not increased with time nor after treatment with adenylate cyclase activators, whereas villous cytotrophoblasts spontaneously demonstrated progressive, increased 3 beta-HSD expression. Moreover, 3 beta-HSD synthesis appeared to precede morphologic syncytial formation. Thus high steroidogenic enzyme expression in placenta is not necessarily closely linked to formation of syncytia. Both Western immunoblot and enzymic activity analyses also indicated that the 3 beta-HSD expressed in these cytotrophoblastic populations was the 3 beta-HSD type I gene product (M(r), 45K) and not 3 beta-HSD type II (M(r), 44K) expressed in fetal testis. In cultures of fetal zone and definitive zone cell of human fetal adrenal, 3 beta-HSD expression was not detected until ACTH was added. ACTH, likely acting in a cyclic AMP-dependent process, induced 3 beta-HSD type II activity and mRNA expression. The higher level of 3 beta-HSD mRNA in definitive zone compared with fetal zone cells was associated with parallel increases in cortisol secretion relative to dehydroepiandrosterone sulfate formation.
...
PMID:Regulation of expression of the 3 beta-hydroxysteroid dehydrogenases of human placenta and fetal adrenal. 827 30


1

---