PACAP-preferring receptors in human hyperplastic prostate.
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Adenylyl Cyclases↗
Biomedical subjects
J C Prieto
Publications and source records attributed to J C Prieto.
Modulation of cyclic AMP and inositol phosphate production in rat prostatic cultures by VIP/PACAP, ATP, and carbachol: role in prostatic proliferation.
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Adenosine Triphosphate↗
Characterization of adenylyl cyclase stimulated by VIP in rat and mouse peritoneal macrophage membranes.
Vasoactive intestinal peptide (VIP) stimulated adenylyl cyclase activity in rat and mouse peritoneal macrophage membranes. GTP potentiated the stimulatory effect of VIP so that it was routinely included at 10 microM GTP. Other agents like GTP, Gpp(NH)p, GTP-gamma-S, sodium fluoride, and forskolin, at a concentration of 0.1 mM, increased the basal activity of enzyme by 3.1, 5.7, 4.7, 3.6, and 7.8-fold, respectively. The stimulation of adenylyl cyclase by VIP was time, temperature, and membrane concentration dependent. Half-maximal enzyme activation (ED50) was very similar in rat and mouse peritoneal macrophage membranes (1.5 +/- 0.1 nM and 1.0 +/- 0.1 nM, respectively). However, VIP showed more efficacy in mouse macrophages membranes (about 3.1-fold basal values) than that in rat macrophage membranes (about 2.5-fold basal values). The relative potency of several peptides upon stimulation of adenylyl cyclase activity showed the following potency in both species: VIP = PACAP38 = PACAP27 > helodermin > PHI > secretin. On the other hand, a M(r)-45 kDa alpha s subunit of Gs protein was demonstrated by both ADP-ribosylation and immunoblot in mouse and rat peritoneal macrophage membranes. The present results, together other previous, strongly suggest that VIP play an important role in the regulation of macrophage function.
Adenosine Diphosphate Ribose↗
Different sites of action of arachidonic acid on steroidogenesis in rat Leydig cells.
The present study in purified rat Leydig cells shows that arachidonic acid may act as an intratesticular factor regulating LH-mediated testicular steroidogenesis. Arachidonic acid decreased, in a dose-dependent manner, the LH-stimulated cAMP and testosterone levels, over 2 h incubation. Incubation of Leydig cells with arachidonic acid did not modify 125I-hCG binding to the cells as compared to control, showing that the action of arachidonic acid is not related to a decrease of hCG binding to the cells. Forskolin-stimulated cAMP and testosterone production were inhibited by 51.65 and 70.9%, respectively, in the presence of arachidonic acid (100 microM), although the ED50 for the diterpene was not changed. When isobutyl-methyl-xanthine was added to the incubation medium, the same percentage of inhibition was found indicating that arachidonic acid inhibition of cAMP production is not due to stimulation of Leydig cell phosphodiesterase activity. Pretreatment of the cells with pertussis toxin, to inactivate Gi, was also without effect on arachidonic acid inhibition of LH-stimulated cAMP production, but pertussis toxin abolished the inhibitory effects of arachidonic acid when adenylate cyclase was stimulated with forskolin. However, arachidonic acid addition resulted in inhibition of LH- and forskolin-stimulated testosterone production, even if the cells were pretreated with pertussis toxin. It can be concluded that: (1) The inhibitory effect of arachidonic acid is neither due to a decrease of hCG binding to Leydig cells nor to a stimulation of cell phosphodiesterase activity; (2) arachidonic acid modulates cAMP production at two different levels, either by activation of Gi protein and by inhibition of Gs protein or adenylate cyclase; (3) the effect of arachidonic acid on steroidogenesis is also beyond cAMP formation.
1-Methyl-3-isobutylxanthine↗
Treatment of actinic prurigo in Chimila Indians.
BACKGROUND: Actinic prurigo has a high prevalence in women of child-bearing age. Its treatment has been, among others, with thalidomide. To avoid the deleterious effects of this drug on the embryo, therapeutic alternatives have been sought. Among these, tetracycline and vitamin E have been investigated as to their influence on the symptoms of actinic prurigo. Both these drugs affect superoxide radicals that are thought to be involved in the pathogenesis of actinic prurigo. MATERIALS AND METHODS: Patients (Chimila Indians with a high prevalence of actinic prurigo) received either (a) tetracycline, 500 mg three times daily, for 6 months, or (b) vitamin E, 100 IU daily, for 6 months. The patients were seen once monthly. There were eight patients in each group. RESULTS: Both drugs used were effective. Pruritus was remarkably improved by either treatment. None of the side effects were severe enough to lead to interruption of treatment, but the observation period posttreatment was relatively short, 4 months for tetracycline and 2 months for vitamin E. The improvement occurred in spite of the continuation of extensive exposure to the sun. CONCLUSIONS: Tetracycline and vitamin E are efficacious in relieving the pruritus of actinic prurigo. Preliminary trials of a combination treatment with these two drugs is a new avenue which has shown in preliminary trials to yield synergistic effects which might allow the dosage of tetracycline to be reduced.
Anti-Bacterial Agents↗
Characterization of vasoactive intestinal peptide/pituitary adenylate cyclase-activating peptide receptors in human benign hyperplastic prostate.
Vasoactive intestinal peptide (VIP) is an important member of the group of neuropeptides that appears to be involved in the regulation of prostatic growth and function. Here we studied VIP receptors in membranes from human benign hyperplastic prostate. Accordingly to observations in rat prostatic membranes, [125I]VIP binding to human prostatic membranes suggested two classes of binding sites with high Kd = 0.22 nM) and low (Kd = 37.7 nM) affinities. VIP bound in human and rat membrane preparations to a common VIP/pituitary adenylate cyclase-activating peptide (PACAP) receptor, as VIP, PACAP-27, and PACAP-38 were equipotent for competition of [125I]VIP binding. A PACAP-preferring receptor appears to be expressed in human prostate, since [125I]PACAP binding was displaced with more potency by PACAP than by VIP, and a messenger RNA corresponding to type I PACAP receptor was found. Cross-linking experiments suggested a VIP receptor of about 71 kDa in human and 52 kDa in rat prostates. The binding of [125I]VIP to membranes and the labeling of the bands observed after electrophoresis were competitively inhibited by GTP, suggesting the coupling of VIP receptors to a G protein. Moreover, after solubilization and cross-linking, we observed a 120-kDa band that corresponded to the VIP receptor-alpha s association. VIP stimulated adenylyl cyclase activity in a dose-dependent manner, but the potency and/or the efficacy of VIP were lower in all human preparations studied than in rat prostatic membranes. In conclusion, this study clearly demonstrates the expression of VIP/PACAP common receptors associated with alpha s protein in human prostate and suggests that these neuropeptides could play an important and complex role in the physiology and pathophysiology of this human gland.
Adenylyl Cyclases↗
[Acute myocardial infarction in Chile: differences between men and women in its evolution and prognosis. Multicenter Study Group on Infarction] (GEMI)].
BACKGROUND: Gender may be a prognostic factor for the evolution of acute myocardial infarction and women may have higher mortality and complication rates. AIM: To study if there are differences in the evolution of acute myocardial infarction between men and women. PATIENT AND METHODS: We have recorded information on risk factors, clinical evolution, treatment and complications of 2,052 patients hospitalized for acute myocardial infarction in 36 Chilean hospitals. The odds ratio for female sex and mortality was calculated using a logistic regression analysis adjusted for risk factors, treatment, invasive procedures and complications. RESULTS: Twenty six percent of analyzed patients were female. Mortality rates among females and males were 11.8 and 20.2% respectively (p < 0.01). Women had higher frequency of smoking, diabetes, obesity and hypertension. Blood lipid levels were similar in both sexes. Compared to men, a lesser proportion of women were treated with thrombolytic agents (25 and 35% respectively), intravenous heparin (54 and 61% respectively), beta blockers (31 and 42% respectively) and intravenous nitrates (53 and 61% respectively). Also, women were subjected to less invasive procedures. The odds ratio for mortality and sex was 1.72 (confidence interval from 1.13 to 2.62). CONCLUSIONS: Female sex is an independent risk factor for acute myocardial infarction mortality.
Adult↗
[Prognostic value of noninvasive makers of coronary reperfusion in patients with acute myocardial infarction treated with thrombolysis].
BACKGROUND: The immediate prognosis of patients with acute myocardial infarction treated with thrombolysis primarily depends on obtaining a satisfactory coronary reperfusion. AIM: To assess the prognostic power of four markers of coronary artery patency in patients with acute myocardial infarction treated with Streptokinase 1.5 million U within the first six hours of symptoms. PATIENTS AND METHODS: In 807 consecutive patients from the Chilean National Registry of Acute Myocardial Infarction we analyzed the resolution of chest pain and ST segment elevation over 50% within the first 90 min, abrupt CK rise within 8 h and T wave inversion in infarct related EKG leads within the first 24 h after thrombolysis. RESULTS: Global in-hospital mortality was 12.1%. Mortality of patients with the presence of 3 or 4 markers of coronary artery patency was 5.1%, in those with resolution of ST elevation and abrupt CK rise was 6.25% and in those with T wave inversion it was 3.9% (p < 0.001). Multivariate analysis, adjusted by age, gender, risk factors, Killip class and infarct location showed that early T wave inversion was the better predictor of a low in-hospital mortality and that its combination with other markers of coronary artery patency did not increase its prognostic power. Early CK rise and the presence of 3 out of 4 reperfusion criteria were also independent predictors of a low mortality. CONCLUSIONS: Non invasive markers of coronary artery patency are associated with a lower in-hospital mortality and may serve as surrogate end points in clinical trials.
Female↗
Protein kinase C regulation of the adenylyl cyclase system in rat prostatic epithelium.
In the context of the crosstalk between transmembrane signalling pathways, we studied the loci within the stimulatory receptor/Gs protein/adenylyl cyclase system at which protein kinase C (PKC) exerts regulatory effects in rat prostatic epithelial cells. The treatment of cells with the PKC activator phorbol 12-myristate 13-acetate (PMA) resulted in an impairment of the stimulation of adenylyl cyclase activity in terms of both potency, as seen with both vasoactive intestinal peptide (VIP) and pituitary adenylyl cyclase-activating peptide (PACAP-27), and efficacy, as seen with the beta-adrenergic agonist isoproterenol. This inhibitory effect of PMA could be prevented by cell incubation with pertussis toxin but not with cholera toxin, pointing to a Gi- but not Gs-dependent mechanism. This hypothesis was reinforced by ADP-ribosylation experiments that showed a low extent of alpha i with pertussis toxin but no change of alpha s with cholera toxin, as well as by the observation of the loss of the ability of low Gpp[NH]p doses to inhibit forskolin-stimulated adenylyl cyclase activity (a measure of Gi function) after cell treatment with PMA. However, the phorbol ester did not modify the adenylyl cyclase catalytic subunit, as shown by experiments on direct stimulation of the enzyme by forskolin. Whatever the exact mechanisms, the results support a crosstalk between the PKC and the adenylyl cyclase systems in rat prostatic epithelial cells in terms of an impairment of adenylyl cyclase stimulation, due presumably to phosphorylation of both membrane receptors (coupled to Gs) and Gi protein, but not of Gs protein or the adenylyl cyclase itself.
Adenosine Diphosphate↗
Protein kinase C isozymes in prostatic epithelial cells from normal, diabetic and insulin-treated diabetic rats.
1. Immunoblot experiments in rat prostatic epithelium using a non-selective antibody against protein kinase C (PKC) allowed to detect three PKC subspecies of 87.5, 55.5 and 34.6 kDa that showed higher, similar and lower immunoreactivity in the membrane than in the cytosolic compartment, respectively. 2. Specific monoclonal antisera revealed that the PKC-gamma isozyme is not expressed in the rat prostatic epithelium, whereas the PKC-beta isozyme was noted only in the cytosolic fraction showing an apparent molecular weight of 75.5 kDa. 3. Induction of diabetes by streptozotocin led to modifications in the expression of PKC isozymes so that the immunoreactivities of the 87.5- and 55.5-kDa PKC forms decreased in both cytosolic and membrane subcellular fractions to different extents. 4. The most important decrease was that of the 55.5-kDa PKC form in cytosol that returned to control values by insulin therapy, whereas PKC-beta suffered also some decrease in diabetes and increased again with insulin treatment.
Animals↗
Alpha-2 adrenoceptors modulate the somatostatinergic system and G protein levels in the rat hippocampus.
alpha 2-Adrenoceptor agonists and somatostatin (SS) exert opposite effects on the spike discharge of pyramidal and granule cells in the rat hippocampus. We studied whether clonidine, an alpha 2-adrenoceptor agonist, and yohimbine, an alpha 2-adrenoceptor antagonist, can modulate somatostatin-like immunoreactivity (SSLI) levels, binding of 125I-Tyr11-somatostatin (125I-Tyr11-SS) to its specific receptors, SS-inhibited adenylyl cyclase (AC) activity, and the guanine-nucleotide binding regulatory proteins Gi and G(o) in the rat hippocampus. Clonidine (1 mg/kg, intraperitoneally (IP) or yohimbine (5 mg/kg, IP) injected at both 10 and 16 hours before decapitation did not affect SSLI content in the hippocampus. Clonidine administration decreased the number of specific SS receptors and increased the apparent affinity in hippocampal membranes. This change in SS binding was not the result of a direct effect of clonidine on these receptors because no effect in binding was produced by high concentrations of clonidine (10(-5) M) when added in vitro. Pretreatment with yohimbine prevented the clonidine-induced in SS binding. Yohimbine alone produced a significant increase in the number of 125I-Tyr11-SS receptors and a decrease in its apparent affinity. Clonidine decreased the ADP-ribosylation of a 41- and a 39-kDa G-protein by pertussis toxin (PTX), whereas yohimbine had no effect on the PTX-catalyzed ADP-ribosylation. No significant differences were seen for the basal or for the forskolin (FK)-stimulated AC enzyme activities in the control, clonidine- and/or yohimbine-treated groups. Somatostatin caused a significantly lower inhibition in AC activity in hippocampal membranes of clonidine-treated rats, whereas yohimbine led to an opposite effect.(ABSTRACT TRUNCATED AT 250 WORDS)
Adenosine Diphosphate↗
Receptors for pituitary adenylate cyclase-activating peptide in human liver.
The presence as well as the pharmacological, molecular, and functional properties of pituitary adenylate cyclase-activating peptide (PACAP) receptors have been analyzed in human liver membranes compared in parallel with vasoactive intestinal peptide (VIP) receptors. [125I]PACAP-27 bound to two classes of receptors with high [dissociation constant (Kd) = 0.47 nmol/L] and low (Kd = 8.0 nmol/L) affinities that represented about 34% and 66% of total binding sites, respectively. The pharmacological profile of [125I]VIP and [125I]PACAP-27 binding to membranes supported the coexistence with VIP receptors of specific receptors for PACAP with a mol wt equal to 67.7K. When [125I]PACAP-27 was used, the order of potency of various related peptides for competition of tracer binding was PACAP-27 greater than PACAP-38 greater than VIP. Both PACAP-27 and VIP stimulated adenylate cyclase activity with similar efficacy, although PACAP-27 showed a potency (half-maximal efficient concentration or EC50 = 0.5 nmol/L) greater than that of VIP (EC50 = 4.1 nmol/L). When the two peptides were present simultaneously in the incubation medium, no additive effect on the stimulation of adenylate cyclase activity was observed, which suggests a unique receptor coupled to this enzyme.
Adenylyl Cyclases↗
The beta 2 receptor plays a role in the hypotensive effect of L-dopa.
Levodopa is a dopamine precursor exerting its cardiovascular effects by conversion to dopamine and by other mechanisms not elucidated yet. Since beta receptors are involved in levodopa Central Nervous System actions, we tried to find out whether adrenergic receptors are involved in the hypotensive effect of levodopa at peripheral level. Experiments were performed on urethane anesthetized (1 ml/100 g i.p.) and heparinized (300 U.I./kg i.v.) rats. Heart rate and mean carotid blood pressure were registered through a polygraph. Pulmonary ventilation was facilitated by a tracheal cannula. Drugs were infused into the femoral vein through a polyethylene catheter. Two series of the following drugs were assayed: a) Dopamine (3.12 to 25.0 micrograms/100 g i.v.) and levodopa (12.5 to 100 micrograms/100 g i.v.) as control groups. b) The aforementioned drugs and dosages were assayed 20 minutes after intravenous propranolol (50 micrograms/100 g), atenolol (100 micrograms/100 g) and ICI 118.55 (25 micrograms/100 g). Dopamine produced a dose-related hypotensive effect (p < 0.02) which was not modified by propranolol, atenolol or ICI 118.55. Levodopa produced a dose-related blood pressure fall which was not changed by atenolol pre-treatment. Contrariwise, propranolol and ICI 118.55 evoked a blood pressure raise with levodopa. Since propranolol and ICI 118.55 have in common the beta 2 receptor blockade, it is suggested that such receptor has a key role in the hypotensive response induced by levodopa.
Animals↗
Characterization of vasoactive intestinal peptide receptors in human liver.
The stoichiometric, pharmacological and molecular properties of vasoactive intestinal peptide (VIP) receptors have been analyzed in human liver membranes and compared in parallel with those in rat liver membranes. The binding of [125I]VIP was rapid, saturable and specific. The stoichiometric data indicated the presence of two classes of binding sites in both human and rat liver membranes with Kd values of 0.22 (human) and 0.20 (rat) nM for the high-affinity site, and 27.3 (human) and 3.6 (rat) nM for the low-affinity site. Tracer binding was displaced by structurally related peptides with an order of potency: VIP = PACAP-27 > helodermin > secretin in human liver, and VIP = PACAP-27 = helodermin > secretin in rat liver. GTP inhibited [125I]VIP binding in a dose-dependent manner suggesting the involvement of a G protein in the signal transduction pathway. Cross-linking experiments revealed an apparent molecular mass for the VIP-receptor complex that was 67,500 +/- 2700 and 50,500 +/- 900 in human and rat preparations, respectively. VIP receptors were functional, since VIP stimulated adenylyl cyclase activity in a dose dependent manner with similar efficacy but different potency in human (ED50 = 1.2 nM) and rat (ED50 = 5.8 nM) liver membranes.
Adenylyl Cyclases↗
G proteins in rat liver proliferation during cholestasis.
Liver proliferation appears to be dually regulated, in part by cyclic AMP levels. Here we studied the alterations in the stimulatory action of cholera toxin and other agents on the adenylyl cyclase system, as well as the status of Gs and Gi protein subunits during the liver proliferation that follows bile duct ligation in rats. The stimulatory effects of glucagon and vasoactive intestinal peptide (which act through membrane receptors) or guanosine 5'-[beta gamma-imido]triphosphate (which interacts with G proteins) and forskolin (which directly activates the adenylyl cyclase catalytic subunit) on liver adenylyl cyclase activity were blunted in cholestasis. The results indicated an impairment in the stimulatory interaction between the alpha s subunit of Gs protein and the adenylyl cyclase catalytic subunit. Indeed, we observed an important decrease in the stimulation of adenylyl cyclase activity by cholera toxin in cholestasis that was accompanied by a reduced extent of [32P]ADP ribosylation of alpha s protein catalyzed by cholera toxin, as revealed by the poor labeling of the 42,000 Da band in liver membranes from cholestatic rats. However, there was no change in the amount of alpha s or beta proteins as measured with immunoblotting techniques. Experiments on [32P]ADP ribosylation of alpha i subunits of Gi proteins indicated an impairment in liver membranes from cholestatic rats, whereas Western blotting for the detection of alpha i subunits showed decreased alpha i3 and increased alpha i2 levels in this condition. Further efforts are needed to better understand the molecular mechanisms underlying the relationship between the observed divergent expression of Gs and Gi proteins and liver cell proliferation in the cholestatic liver.
Adenosine Diphosphate↗
Role of lindane in membranes. Effects on membrane fluidity and activity of membrane-bound proteins.
The influence of lindane (gamma-hexachlorocyclohexane) on fluidity of plasma membranes from rat renal cortical tubules has been investigated. Preincubation with lindane increased membrane fluidity. This effect was accompanied by (i) a decrease in the transport of glucose with regard to the controls and (ii) an inhibition of the beta-adrenergic stimulatory activity upon cyclic AMP accumulation. However, a significant decrease of the membrane fluidity was found when rats were injected with lindane for 12 days. The injection of lindane exerted the opposite effect on the membrane proteins, the glucose transporter and the beta-adrenergic receptor, enhancing the glucose uptake and increasing the isoproterenol-stimulated cycle AMP accumulation. A possible explanation of the difference could involve a resistance to membrane disordering by lindane through a regulatory mechanism that would balance the activity of many lindane-sensitive proteins in insecticide-injected rats.
Animals↗
Effect of sulpiride on somatostatin receptors, somatostatin-like immunoreactivity and modulation of adenylyl cyclase activity in the rat brain.
The present study investigates the effects of the administration of an intracerebroventricular (i.c.v.) dose of 500 micrograms/rat of the neuroleptic (-) sulpiride on somatostatin-like immunoreactivity (SSLI) levels, 125I-Tyr11-SS binding to its specific receptors, SS-modulated adenylyl cyclase (AC) activity and the pertussis toxin (PTX) substrates measured by toxin-catalysed ADP ribosylation of the alpha-subunits from G-proteins. (-) Sulpiride significantly decreased the SSLI levels in the frontoparietal cortex at 30 min but was without effect on the SSLI concentration in the striatum. This decrease had disappeared within 24 hr. The administration of (-) sulpiride produced a significant increase in the number of 125I-Tyr11-SS receptors and a significant reduction in their affinity at 30 min after injection in the striatum without affecting the frontoparietal cortex. The effects of the (-) sulpiride injection had disappeared after 24 hr. This change in SS binding was not due to a direct effect of (-) sulpiride on these receptors since no effect on binding was produced by high concentrations of (-) sulpiride (10(-5) M) when added in vitro. No significant differences were seen in either brain region for the basal or the forskolin (FK)-stimulated AC enzyme activities in the control and (-) sulpiride groups. In the (-) sulpiride group, the capacity of SS to inhibit FK-stimulated AC in the frontoparietal cortex was significantly higher than in the control group with no significant difference in the striatum.(ABSTRACT TRUNCATED AT 250 WORDS)
Adenosine Diphosphate Ribose↗
Neuropeptide Y inhibits vasoactive intestinal peptide-stimulated adenylyl cyclase in rat ventral prostate.
Neuropeptide Y (NPY), a peptide present in the prostate gland, was found to inhibit vasoactive intestinal peptide (VIP)-stimulated cyclic AMP accumulation in isolated rat prostatic epithelial cells as well as VIP-stimulated adenylyl cyclase activity in rat prostatic membranes. The inhibitory effect of NPY was selective for the VIP receptor/effector system since it was also observed when using pituitary adenylyl cyclase activating peptide (PACAP-27) which presumably recognizes VIP receptors in this gland, but not when using unrelated substances such as isoproterenol or forskolin. NPY did not modify either the general lipid membrane microviscosity or the VIP-receptor binding. The inhibitory effect of VIP was blocked by pretreatment of the prostatic membranes with pertussis toxin. These results suggest the presence of NPY receptors in rat ventral prostate coupled in an inhibitory manner to adenylyl cyclase through a guanine nucleotide regulatory Gi protein.
Adenylyl Cyclase Inhibitors↗