Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: UNIPROT:P01189 (beta-endorphin)
 21,003 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have investigated the effect of the proteinase inhibitors 1,10-phenantroline (OP) and phenylmethylsulfonyl fluoride (PMSF) on steroidogenesis in rat adrenal cortex. Both PMSF and OP inhibited adrenocorticotropin (ACTH)- and 8-Br cAMP-induced stimulation of corticosterone synthesis. On the contrary, arachidonic acid-induced stimulation of corticosterone synthesis was only slightly inhibited by PMSF and unchanged by OP. Intra- and extracellular cAMP levels were determined by radioimmunoassay. While PMSF did not affect neither the intra- nor the extracellular cAMP levels, OP decreased the intra- and extracellular levels of unstimulated as well as ACTH-stimulated cells. The site of action of the proteinase inhibitors was also studied by recombination of mitochondria with the different subcellular fractions in vitro. Addition of PMSF abolished the stimulation achieved by in vitro activation of cytosol by cAMP and PKA. On the other hand, OP completely inhibited the activation of mitochondria. Our results provide evidence for the involvement of proteinases in ACTH-induced stimulation of steroidogenesis in adrenal cortex both prior to the release of arachidonic acid and at the level of cholesterol transport from the outer to the inner mitochondrial membrane.
 ...
PMID:Site of action of proteinases in the activation of steroidogenesis in rat adrenal gland. 859 3

Adrenocorticotropic hormone (ACTH) triggers well-defined responses in Y-1 cells. Among them is steroidogenesis stimulation. We have previously shown that phorbol 12-myristate 13-acetate (PMA), an activator of the calcium- and phospholipid-dependent protein kinase (PKC) is able to mimic all the responses triggered by ACTH in these cells, including steroidogenesis stimulation. Short (2 h) treatment with PMA leads to only 20-30% of the maximal steroidogenesis stimulation obtained with ACTH. However, the steroid secretion in the 2 h that follows the short-term (2 h) PMA treatment reaches the same levels as observed with ACTH, i.e., a 12- to 15-fold increase. We also show that this effect is restricted to cells treated with PMA for up to 4 h, while treatment for longer periods of time causes a reduction of the steroid biosynthesis rate, an effect that is not observed in cells treated with ACTH or N6,2'-0-dibutyryladenosine 3',5'-cyclic monophosphate (dcAMP). These results suggest that activation of PKC can elicit the first phase of ACTH steroidogenesis stimulation, but not the second one, which strictly depends on activation of cAMP-dependent protein kinase.
 ...
PMID:Patterns of long-term steroidogenesis stimulation by ACTH and phorbol ester. 873 27

The aim of this study was to determine the ability of corticotropin-releasing hormone (CRH), lysine vasopressin (LVP), oxytocin (OT), and angiotensin II (AII) to stimulate adrenocorticotropin (ACTH) secretion from porcine anterior pituitary (AP) cells in vitro and to evaluate the role of protein kinase C (PKC) in the interaction between CRH and LVP. In this study, porcine AP cells were enzymatically and mechanically dispersed, cultured (150,000 cells/well) for 4 d, and then challenged with doses of various neuropeptides for 3 hr. CRH (10(-7)-10(-10) M) was the most potent of the peptides tested in stimulating ACTH release from porcine AP cells. In fact, none of the other peptides consistently affected ACTH concentrations relative to basal levels. However, LVP potentiated CRH action, even though by itself, it failed to stimulate ACTH production. Neither OT or AII potentiated CRH-stimulated ACTH release from porcine AP cells. To determine whether the inter-action between CRH and LVP was regulated partially by the protein Kinase C (PKC) pathway, we challenged AP cells in a 30-min incubation with 10(-7) M staurosporine (ST), a treatment predicted to decrease PKC activity. Then, cells were washed and challenged with 10(-9) M LVP, 10(-9) M CRH, and 10(-9) M CRH + LVP. Treatment with ST decreased (P < 0.05) CRH + LVP-stimulated ACTH release. To further demonstrate an interaction between protein kinase A (PKA) and PKC transduction pathways in the observed synergism between CRH and LVP to enhance ACTH secretion, we also challenged AP cells with 10(-7) M phorbol 12, 13-myristate acetate (PMA) and 5 microM forskolin (FOR) for 3 hr. This treatment was predicted to enhance PKA and PKC activities, respectively, and thereby enhance ACTH concentrations. Challenging cells with FOR + PMA enhanced (P < 0.001) ACTH release above basal concentrations, but more important, it increased (P < 0.001) ACTH concentration above that elicited by either drug given alone. Taken together, our in vitro studies support the conclusion that CRH is the principal regulator of ACTH secretion in the pig. In contrast to the results in most other species evaluated, vasopressin alone did not affect ACTH release. However, LVP can enhance the effectiveness of CRH in releasing ACTH, and this enhancement appears to rely, at least in part, on the activation of the PKC signal transduction pathway.
 ...
PMID:Effects of corticotropin-releasing hormone, lysine vasopressin, oxytocin, and angiotensin II on adrenocorticotropin secretion from porcine anterior pituitary cells. 873 67

The effects of various neuropeptides on human plasma cells were studied. Of the various neuropeptides tested, vasoactive intestinal peptide (VIP) enhanced Ig production and growth in human plasma cell lines, IM-9 and AF-10, and in plasma cells generated in vivo (four out of four patients with plasma cell leukemia) and in vitro. In contrast, other neuropeptides (neuropeptide Y, somatostatin, substance P, peptide YY, neurokinin A, calcitonin gene-related peptide, chole-cystokinin octapeptide, and beta-endorphin) were ineffective. Moreover, VIP-induced enhancement was specifically blocked by VIP receptor antagonist. Among the various cytokines, IL-6, GH, and insulin-like growth factor I (IGF-I) also enhanced Ig production and thymidine uptake in plasma cells. However, VIP-induced enhancement was not mediated by IL-6, GH, or IGF-I because antibodies to these cytokines failed to block VIP-induced enhancement. Phorbol 12,13 dibutyrate enhanced Ig production and thymidine uptake in plasma cells, and the Phorbol 12,13 dibutyrate-induced enhancement was blocked by H7 (a protein kinase C inhibitor) but not by H8 (a protein kinase A inhibitor). Similarly, VIP-induced enhancement was blocked by H7 but not by H8. Collectively, VIP enhances plasma cell responses via mechanisms that may involve protein kinase C.
 ...
PMID:Vasoactive intestinal peptide enhances immunoglobulin production and growth in human plasma cells via mechanisms that may involve protein kinase C. 876 69

Besides acting as an important cofactor in the biosynthesis of catecholamine, ascorbic acid (AA) also modulates the activity of peptidylglycine-alpha-amidating monooxygenase for the post-translational modification of neuropeptides such as alpha-MSH and TRH. We report here a novel action of AA in modulating the secretion of immunoreactive beta-endorphin (ir-beta EP) and mRNA expression of proopiomelanocortin (POMC) following the activation of cAMP-dependent protein kinase A pathway in rat hypothalamic neurons. Primary cultures of hypothalamic neurons from neonatal rats as previously described were employed in the present studies. Six days after plating, cultures were replenished with serum-free media and incubated with vehicle or various doses of AA in the presence or absence of forskolin, 3-isobutyl-1-methylxanthine (IBMX), N6,2'-O-dibutyryladenosine 3'5'-(cyclic)monophosphate [(Bu)2cAMP]. Whereas the basal ir-beta EP release was 22.0 +/- 0.4 pg/well (mean +/- S.E.; n = 3), 10 microM of forskolin treatment increased ir-beta EP release approximately 4.2-fold. Co-incubation with AA enhanced forskolin induced ir-beta EP release and that this enhancing effect of AA was both time related and dose-dependent, with an ED50 of approximately 10 microM and an Emax of 100 microM. At the concentration of 10 microM, AA augmented ir-beta EP release approximately 6.1-fold that of cultures treated with forskolin alone. A similar potentiating effect of AA was also seen in cultures co-treated with IBMX or with (Bu)2cAMP. These enhancing effects of AA were similarly found in the abundance of total cAMP and of POMC mRNA of cultures which received identical treatments. However, it is important to point out that AA alone did not modulate ir-beta EP release or the abundance of POMC mRNA or total cAMP levels of the hypothalamic cultures when protein kinase A pathway was not activated. We thus conclude that AA augments cAMP-dependent protein kinase A pathway-induced production and release of beta EP from rat hypothalamic neurons in culture. Furthermore, this biological effect of AA is, at least in part, mediated through enhancing the responsiveness of the adenylyl cyclase-cAMP system.
 ...
PMID:Ascorbic acid augments the adenylyl cyclase-cAMP system mediated POMC mRNA expression and beta-endorphin secretion from hypothalamic neurons in culture. 882 63

Mifepristone (RU486), bovine corticotropin-releasing hormone (CRH), arginine vasopressin (VP), adrenocorticotropin (ACTH1-24), and protein kinase activators (forskolin, [FSK]; phorbol 12-myristate 13-acetate [PMA]) were used in vitro to investigate their direct effect on adrenocorticosteroidogenesis. Bovine adrenocortical fasciculata/reticularis cells (2 x 10(5) viable cells/well) were cultured for 3 d in medium supplemented with 10% fetal calf serum. After incubation for an additional 24 hr in serum-free medium, cells were treated with serum-free medium alone (Control) or various concentrations of ACTH, CRH, VP, FSK, PMA, RU486, and/or various concentrations for 1, 2, 4, or 24 hr. Medium content of cortisol and progesterone were determined by radioimmunoassays. ACTH, CRH, FSK, and PMA each stimulated (P < 0.05) secretion of cortisol in time- and dose-related manners. Although these agents stimulated (P < 0.05) secretion of progesterone in a dose-related manner, medium content of progesterone declined (P < 0.05) over time. The minimal effective doses of ACTH and CRH required to stimulate (P < 0.05) secretion of cortisol relative to the Control over a 4-hr culture period were 0.01 nM and 3 nM, respectively. Relative to observations at 1 hr posttreatment, 24-hr treatment with ACTH or CRH increased the medium content of cortisol by an additional 19.8- and 48-fold, respectively (whereas content of progesterone declined over that time period). VP-stimulated secretion of cortisol was time- (P < 0.05) but not dose-related. Specifically, by 24-hr posttreatment, the medium content of cortisol was increased (P < 0.05) 4.6-fold relative to the quantity of cortisol secreted by 1-hr postaddition of VP (0.01 to 1 microM). Co-treatment with RU486 (1 microM) decreased (p < 0.05) FSK-, ACTH- and CRH-stimulated secretion of cortisol by 77, 27, and 56%, respectively. Similarly, the stimulatory effects of ACTH and CRH on progesterone secretion were reduced (P < 0.05) by 40 and 22%, respectively, by co-addition of RU486. The inhibitory action of RU486 on production of cortisol was no longer apparent by 24 hr after treatment. These observations indicate that RU486 can act as a steroid agonist and as well as an antagonist. These data characterize time- and dose-related direct actions of ACTH, CRH, and RU486 on adrenocorticosteroidogenesis. This information will assist efforts to clarify complex intra-adrenal interactions of neurohormones, growth factors, and endogenous steroids.
 ...
PMID:Mifepristone modulation of ACTH and CRH regulation of bovine adrenocorticosteroidogenesis in vitro. 883 27

Bovine adrenal zona fasciculata (AZF) cells express a noninactivating K+ current (IAC) that is inhibited by adrenocorticotropic hormone (ACTH) at picomolar concentrations. Inhibition of IAC may be a critical step in depolarization-dependent Ca2+ entry leading to cortisol secretion. In whole-cell patch clamp recordings from AZF cells, we have characterized properties of IAC and the signalling pathway by which ACTH inhibits this current. IAC was identified as a voltage-gated, outwardly rectifying, K(+)-selective current whose inhibition by ACTH required activation of a pertussis toxin-insensitive GTP binding protein. IAC was selectively inhibited by the cAMP analogue 8-(4-chlorophenylthio)-adenosine 3':5'-cyclic monophosphate (8-pcpt-cAMP) with an IC50 of 160 microM. The adenylate cyclase activator forskolin (2.5 microM) also reduced IAC by 92 +/- 4.7%. Inhibition of IAC by ACTH, 8-pcpt-cAMP and forskolin was not prevented by the cAMP-dependent protein kinase inhibitors H-89 (5 microM), cAMP-dependent protein kinase inhibitor peptide (PKI[5-24]) (2 microM), (Rp)-cAMPS (500 microM), or by the nonspecific protein kinase inhibitor staurosporine (100 nM) applied externally or intracellularly through the patch pipette. At the same concentrations, these kinase inhibitors abolished 8-pcpt-cAMP-stimulated A-kinase activity in AZF cell extracts. In intact AZF cells, 8-pcpt-cAMP activated A-kinase with an EC50 of 77 nM, a concentration 2,000-fold lower than that inhibiting IAC half maximally. The active catalytic subunit of A-kinase applied intracellularly through the recording pipette failed to alter functional expression of IAC. The inhibition of IAC by ACTH and 8-pcpt-cAMP was eliminated by substituting the nonhydrolyzable ATP analogue AMP-PNP for ATP in the pipette solution. Penfluridol, an antagonist of T-type Ca2+ channels inhibited 8-pcpt-cAMP-induced cortisol secretion with an IC50 of 0.33 microM, a concentration that effectively blocks Ca2+ channel in these cells. These results demonstrate that IAC is a K(+)-selective current whose gating is controlled by an unusual combination of metabolic factors and membrane voltage. IAC may be the first example of an ionic current that is inhibited by cAMP through an A-kinase-independent mechanism. The A-kinase-independent inhibition of IAC by ACTH and cAMP through a mechanism requiring ATP hydrolysis appears to be a unique form of channel modulation. These findings suggest a model for cortisol secretion wherein cAMP combines with two separate effectors to activate parallel steroidogenic signalling pathways. These include the traditional A-kinase-dependent signalling cascade and a novel pathway wherein cAMP binding to IAC K+ channels leads to membrane depolarization and Ca2+ entry. The simultaneous activation of A-kinase- and Ca(2+)-dependent pathways produces the full steroidogenic response.
 ...
PMID:Adrenocorticotropic hormone and cAMP inhibit noninactivating K+ current in adrenocortical cells by an A-kinase-independent mechanism requiring ATP hydrolysis. 889 75

The mechanisms of corticotropin-releasing hormone (CRH) induced excitation of ACTH-secreting adenoma cells were investigated using the perforated whole-cell clamp technique and intracellular Ca2+ concentration ([Ca2+]i) measurement. CRH depolarized ACTH-secreting adenoma cells by activating a nonselective cation current that showed slight inward rectification. This channel did not seem to be a member of the Ca(2+)-activated cation currents because it was activated even when the [Ca2+]i was chelated below 50 nM. The activation of the current was induced by protein kinase A-mediated pathways. By [Ca2+]i measurement, CRH increased [Ca2+]i of these cells dependently on voltage-gated Ca2+ current. This CRH-induced [Ca2+]i increase was abolished in Na(+)-free extracellular solution, but was not abolished by the addition of 5 microM tetrodotoxin to the extracellular solution. CRH-induced ACTH secretion from the cultured adenoma cells was also abolished in Na(+)-free extracellular solution, but not in tetrodotoxin-containing extracellular solution. These data indicate that a Na+ current (maybe the nonselective cation current) other than voltage-gated Na+ current plays an important role in CRH-induced [Ca2+]i increase and ACTH secretion. CRH also activated a nonselective cation current in nonadenoma human corticotrophs, suggesting that the activation of a nonselective cation current is a physiological mechanism of CRH-induced excitation in human corticotrophs.
 ...
PMID:Corticotropin-releasing hormone excites adrenocorticotropin-secreting human pituitary adenoma cells by activating a nonselective cation current. 890 22

Incubation of rat adrenal glomerulosa cells with low concentrations (up to 50 nM) of the protein kinase (PKC) inhibitor staurosporine (ST) inhibited aldosterone (ALDO) and cyclic AMP (cAMP) production stimulated by adrenocorticotropic hormone (ACTH) and cholera toxin. Only higher concentrations (1.6 microM) of staurosporine inhibited dibutyryl-cAMP- and forskolin-induced stimulation of aldosterone production. cAMP levels were increased only with low concentrations of the PKC inhibitor. This latter increase was avoided by treatment with a maximal concentration of isobutylmethylxanthine (MIX). Our results suggest that: (1) second messengers other than cAMP are involved in ACTH action; (2) staurosporine inhibits different kinases involved in ACTH action in a dose-dependent manner; (3) the protein kinase inhibited by high concentrations of staurosporine appears to be the cAMP-dependent kinase, PKA; and (4) the protein kinase inhibited by low concentrations of staurosporine remains to be identified. This latter species is suggested as being involved in mediating ACTH-induced activation of Gs.
 ...
PMID:Effects of staurosporine on ACTH-mediated stimulation of aldosterone production. 891 88

Intracellular Ca2+ oscillations play an important role in the induction of alpha-MSH release from pituitary melanotrope cells of Xenopus laevis. Oscillatory, secretory and adenylyl cyclase activities are all inhibited by dopamine, neuropeptide Y (NPY) and baclofen (a GABAB receptor agonist) and stimulated by sauvagine. In this study, we test the hypothesis that these neural messengers regulate the Ca2+ oscillations via a cAMP/protein kinase A (PKA)-dependent mechanism. To this end, video-imaging microscopy was applied to single Xenopus melanotropes loaded with the Ca2+ indicator Fura-2. The cAMP-dependent PKA inhibitor H89 blocked Ca2+ oscillations as well as the stimulatory actions of 8-Br-cAMP and sauvagine. Treatment of cells inhibited by baclofen with either 8-Br-cAMP or sauvagine led to a reappearance of Ca2+ oscillations. A similar result was found for cells inhibited by NPY. Neither 8-Br-cAMP nor sauvagine induced Ca2+ oscillations in cells inhibited by dopamine. Depolarizing dopamine-inhibited cells with high potassium also failed to induce oscillations, but combining 8-Br-cAMP with membrane depolarization induced oscillations. It is concluded that sauvagine, baclofen and NPY work primarily through a cAMP/PKA-pathway while dopamine inhibits Ca2+ oscillations in a dual fashion, namely via both a cAMP-dependent and a cAMP-independent mechanism, the latter probably involving membrane hyperpolarization.
 ...
PMID:Calcium oscillations in melanotrope cells of Xenopus laevis are differentially regulated by cAMP-dependent and cAMP-independent mechanisms. 893 52


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>