Lilian Puebla

Dopamine enhances somatostatin receptor-mediated inhibition of adenylate cyclase in rat striatum and hippocampus

Although there is evidence that suggests that dopamine (DA) has stimulatory effects on somatostatinergic transmission, it is unknown to date if DA increases the activity of the somatostatin (SS) receptor‐effector system in the rat brain. In this study, we evaluated the effects of the administration of DA and the DA D1‐like (D1, D5) receptor antagonist SCH 23390 and the D2‐like (D2, D3, D4) receptor antagonist spiperone on the SS receptor‐adenylate cyclase (AC) system in the Sprague‐Dawley rat striatum and hippocampus. An intracerebroventricular injection of DA (0.5 μg/rat) increased the number of SS receptors and decreased their apparent affinity in the striatum and hippocampus 15 hr after its administration. The simultaneous administration of the DA receptor antagonists SCH 23390 (0.25 mg/kg, ip) and spiperone (0.1 mg/kg, ip) before DA injection partially prevented the DA‐induced increase in SS binding. The administration of SCH 23390 plus spiperone alone produced a significant decrease in the number of SS receptors in both brain areas studied at 15 hr after injection, an effect that disappeared at 24 hr. The increased number of SS receptors in the DA‐treated rats was associated with an increased capacity of SS to inhibit basal and forskolin (FK)‐stimulated (AC) activity in the striatum and hippocampus at 15 hr after injection. This effect had disappeared at 24 hr. By contrast, basal and FK‐stimulated enzyme activities were unaltered after DA injection. No significant changes in the levels of the αI (αi1 + αi2) subunits were found in DA‐treated rats as compared with control rats. In addition, the immunodetection of the αi1 or αi2 subunits showed no significant changes in their levels in DA‐treated rats when compared with controls. DA injection also induced an increase in SS‐like immunoreactive content in the rat striatum but not hippocampus at 15 hr after administration and returned to control values at 24 hr. These results provide direct evidence of a functional linkage between the dopaminergic and somatostatinergic systems at the molecular level. J. Neurosci. Res. 48:238–248, 1997.

Modulation of somatostatin receptors, somatostatin content and Gi proteins by substance P in the rat frontoparietal cortex and hippocampus

Substance P (SP) and somatostatin (SRIF) are widely spread throughout the CNS where they play a role as neurotransmitters and/or neuromodulators. A colocalization of both neuropeptides has been demonstrated in several rat brain areas and SP receptors have been detected in rat cortical and hippocampal somatostatinergic cells. The present study was thus undertaken to determine whether SP could modulate SRIF signaling pathways in the rat frontoparietal cortex and hippocampus. A single intraperitoneal injection of SP (50, 250 or 500 µg/kg) induced an increase in the density of SRIF receptors in membranes from the rat frontoparietal cortex at 24 h of its administration, with no change in the hippocampus. The functionality of the SRIF receptors was next investigated. Western blot analysis of Gi proteins demonstrated a significant decrease in Giα1 levels in frontoparietal cortical membranes from rats treated acutely (24 h) with 250 µg/kg of SP, which correlated with a decrease in functional Gi activity, as assessed by use of the non‐hydrolyzable GTP analog 5′‐guanylylimidodiphosphate. SRIF‐mediated inhibition of basal or forskolin‐stimulated adenylyl cyclase activity was also significantly lower in the frontoparietal cortex of the SP‐treated group, with no alterations in the catalytic subunit of the enzyme. SRIF‐like immunoreactivity content was increased in the frontoparietal cortex after acute (24 h) SP administration (250 or 500 µg/kg) as well as in the hippocampus in response to 7 days of SP (250 µg/kg) administration. All these SP‐mediated effects were prevented by pretreatment with the NK1 receptor antagonist RP‐67580. Although the physiologic significance of these results are unknown, the increase in SRIF receptor density together with the desensitization of the SRIF inhibitory signaling pathway might be a mechanism to potentiate the stimulatory pathway of SRIF, inducing a preferential coupling of the receptors to PLC.

Exogenous histamine increases the somatostatin receptor/effector system in the rat frontoparietal cortex

The present study examined the effects of histamine on somatostatin-like immunoreactivity levels, binding of 125I-[Tyr11]somatostatin to its specific receptors, somatostatin inhibition of basal and forskolin-stimulated adenylyl cyclase activity and inhibitory guanine-nucleotide binding protein (Gi) function in the rat frontoparietal cortex. An intracerebroventricular (i.c.v.) dose of 10 micrograms or 1 microgram of histamine induced an increase in the number of specific 125I-[Tyr11]somatostatin receptors (590 +/- 22 vs 358 +/- 12 fmol/mg protein, P < 0.001 and 455 +/- 20 vs. 342 +/- 21 fmol/mg protein, P < 0.01, respectively) together with a decrease in their apparent affinity (0.76 +/- 0.04 vs 0.39 +/- 0.02 nM, P < 0.001 and 0.60 +/- 0.03 vs 0.39 +/- 0.05 nM, P < 0.01, respectively) in rat frontoparietal cortex membranes. This increase in tracer binding was not due to a direct effect of histamine on the somatostatin receptors since no change in binding was produced when histamine was added directly to the incubation medium. No significant differences were seen for either the basal or forskolin-stimulated adenylyl cyclase activity in frontoparietal cortex membranes of histamine-treated rats as compared with the control group. In rats treated with 10 micrograms of histamine, however, somatostatin caused a significantly greater inhibition of basal and forskolin-stimulated adenylyl cyclase activity as compared to the control group (33 +/- 4% vs 19 +/- 1% inhibition, P < 0.05 and 31 +/- 1% vs 21 +/- 3% inhibition, P < 0.05, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of nitric oxide on the somatostatinergic system in the rat exocrine pancreas

Nitric oxide (NO) and somatostatin (SS) are two important mediators of the exocrine and endocrine pancreas, exerting opposite effects on this organ. There is strong evidence suggesting an interaction between pancreatic NO and SS. The aim of this study was to determine whether L-arginine (L-Arg), the substrate for NO synthase (NOS), and Nomega-nitro-L-arginine methyl ester (L-NAME), a NOS inhibitor, regulate pancreatic somatostatin-like immunoreactivity (SSLI) content and the SS mechanism of action in pancreatic acinar cell membranes. L-Arg (150 mg/kg, intraperitoneally (i.p.)), L-NAME (50 mg/kg, i.p.) or L-NAME plus L-Arg were injected twice daily at 8 h intervals for 8 days. L-Arg decreased pancreatic SSLI content as well as the number of SS receptors in pancreatic acinar cell membranes whereas L-NAME increased both parameters. The stable SS analogue SMS 201-995 induced a significantly lower inhibition of forskolin-stimulated adenylyl cyclase activity in pancreatic acinar cell membranes from L-Arg-treated rats whereas an increased inhibition was observed in pancreatic acinar membranes from L-NAME-treated rats. These results indicate that the NO system may contribute to the regulation of the pancreatic somatostatinergic system.

Effect of dimaprit and cimetidine on the somatostatinergic system in the rat frontoparietal cortex

A recent study carried out by this laboratory demonstrated that exogenous histamine increases the somatostatin (SS) receptor/effector system in the rat frontoparietal cortex (Puebla and Arilla, 1995). In the present study we examined the participation of the H2-histaminergic system in this modulation by use of the H2-receptor agonist and antagonist dimaprit and cimetidine, respectively. Dimaprit administration [20 μg/rat, intracerebroventricularly (ICV)] to rats 2 hours before decapitation increased the number of SS receptors in the frontoparietal cortex without changing the affinity constant. Pretreatment with cimetidine (20 μg/rat, ICV) prevented the dimaprit-induced changes in SS binding in the frontoparietal cortex, whereas cimetidine alone (20 μg/rat, ICV) had no observable effect on this parameter. The in vitro addition of dimaprit or cimetidine to frontoparietal cortex membranes from untreated rats did not markedly affect the SS binding characteristics. Somatostatin caused a significantly higher inhibition of basal and forskolin (FK)-stimulated adenylyl cyclase (AC) activity in frontoparietal cortex membranes from dimaprit-treated rats than in controls, an effect that was prevented by pretreatment with cimetidine. No significant differences, however, were detected for the basal or FK-stimulated AC enzyme activity in the control, dimaprit-, and/or cimetidine-treated groups, which suggests no impairment of the AC catalytic subunit. In addition, the functional activity of the guanine nucleotide-binding inhibitory protein Gi, as measured by the capacity of the stable GTP analogue 5′-guanylylimidodiphosphate [Gpp(NH)p] to inhibit FK-stimulated AC activity, was not altered by dimaprit. Thus, the increased SS-mediated inhibition of AC activity observed in the dimaprit-treated rats may be caused by the increase in the number of SS receptors. Neither dimaprit nor cimetidine affected somatostatinlike immunoreactivity (SSLI) content. The present results, together with the fact that SS and histamine have been shown to influence locomotor activity and nociception in a similar manner, suggest that some of the neurotransmitter effects of SS may be modulated by histamine via H2-histaminergic receptors.

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)

Involvement of presynaptic histamine H3 receptors in the modulation of somatostatin binding and its effects on adenylyl cyclase activity in the rat frontoparietal cortex

Abstract: Thioperamide (2 mg/kg, i.p.), a histamine H3‐receptor antagonist, increased the number of somatostatin (SS) receptors, with no change in the affinity constant, in the rat frontoparietal cortex. This effect was prevented by treatment with (R)‐α‐methylhistamine (3.2 mg/kg, i.p.), a histamine H3‐receptor agonist. Thioperamide also induced an increase in SS binding in rats pretreated with mepyramine, a histamine H1‐receptor antagonist, or cimetidine, a histamine H2‐receptor antagonist. Pretreatment with mepyramine plus cimetidine administered simultaneously antagonized the thioperamide effect on SS binding. The increase in the number of SS receptors was accompanied by a greater SS‐mediated inhibition of basal and forskolin‐stimulated adenylyl cyclase (AC) activity in frontoparietal cortical membranes in the thioperamide group. Furthermore, the functional activity of the guanine nucleotide‐binding inhibitory protein (Gi protein) was not altered by thioperamide or (R)‐α‐methylhistamine administration in frontoparietal cortical membranes. In rats treated with mepyramine plus thioperamide or cimetidine plus thioperamide, the increase in the number of SS receptors was also accompanied by an increased SS inhibition of AC activity. Thioperamide induced a significant increase in SS‐like immunoreactivity content in the frontoparietal cortex. Altogether, these results suggest that frontoparietal cortical histamine may play, at least in part, a role in the regulation of the somatostatinergic system.

Young and Especially Senescent Endothelial Microvesicles Produce NADPH: The Fuel for Their Antioxidant Machinery

In a previous study, we demonstrated that endothelial microvesicles (eMVs) have a well-developed enzymatic team involved in reactive oxygen species detoxification. In the present paper, we demonstrate that eMVs can synthesize the reducing power (NAD(P)H) that nourishes this enzymatic team, especially those eMVs derived from senescent human umbilical vein endothelial cells. Moreover, we have demonstrated that the molecules that nourish the enzymatic machinery involved in NAD(P)H synthesis are blood plasma metabolites: lactate, pyruvate, glucose, glycerol, and branched-chain amino acids. Drastic biochemical changes are observed in senescent eMVs to optimize the synthesis of reducing power. Mitochondrial activity is diminished and the glycolytic pathway is modified to increase the activity of the pentose phosphate pathway. Different dehydrogenases involved in NADPH synthesis are also increased. Functional experiments have demonstrated that eMVs can synthesize NADPH. In addition, the existence of NADPH in eMVs was confirmed by mass spectrometry. Multiphoton confocal microscopy images corroborate the synthesis of reducing power in eMVs. In conclusion, our present and previous results demonstrate that eMVs can act as autonomous reactive oxygen species scavengers: they use blood metabolites to synthesize the NADPH that fuels their antioxidant machinery. Moreover, senescent eMVs have a stronger reactive oxygen species scavenging capacity than young eMVs.