Differential alterations in responsiveness in particulate and soluble guanylate cyclases in pregnant guinea pig myometrium.

OBJECTIVE: The mechanism underlying myometrial quiescence during pregnancy is unknown. Our group has previously shown that during pregnancy myometrial cyclic guanosine monophosphate content rises to several hundred times the nonpregnant levels, only to abruptly decline days before the onset of labor. Cyclic guanosine monophosphate plays an integral role in the relaxation of smooth muscle. The aim of this investigation was therefore to determine the effects of pregnancy on both soluble and particulate guanylate cyclase enzymatic activities and messenger ribonucleic acid expressions. STUDY DESIGN: Myometrium was obtained from randomly cycling adult nonpregnant guinea pigs and near-term (50-60 days' gestation) pregnant guinea pigs of similar chronologic age. Subcellular fractions were prepared by differential ultracentrifugation. Guanylate cyclase activity was determined by the conversion of guanosine triphosphate to cyclic guanosine monophosphate under basal or stimulated conditions in either the soluble guanylate cyclase or particulate guanylate cyclase fraction. A nitric oxide donor, S-nitroso- N-penacillamine, was used to activate soluble guanylate cyclase (n = 10 animals in each group). Several natriuretic peptides (atrial natriuretic peptide, brain natriuretic peptide, and C-type natriuretic peptide) and uroguanylin were used to stimulate the different particulate guanylate cyclase isoforms guanylate cyclase A, guanylate cyclase B, and guanylate cyclase C, respectively, in pregnant (n = 8) and nonpregnant (n = 6) animals. Cyclic guanosine monophosphate content was measured by radioimmunoassay, and enzymatic activity was expressed as picomoles of cyclic guanosine monophosphate per milligram of protein per minute. Total guanylate cyclase represented the sum of soluble guanylate cyclase and particulate guanylate cyclase activities for a tissue. To investigate whether the observed changes in guanylate cyclase activity were paralleled by changes in receptor expression, messenger ribonucleic acid levels of the genes for guanylate cyclase A and guanylate cyclase B isoforms were quantified by ribonuclease protection assay (n = 5 animals in each group). RESULTS: Under basal conditions particulate guanylate cyclase represented 78% (nonpregnant state) to 88% (during pregnancy) of the total guanylate cyclase activity in the guinea pig myometrium. Pregnancy further reduced myometrial soluble guanylate cyclase (both basal and stimulated by nitric oxide) relative to the nonpregnant state. Pregnancy selectively increased atrial natriuretic peptide-stimulated particulate guanylate cyclase activity (attributed to guanylate cyclase A), although it did not change basal myometrial particulate guanylate cyclase activity in general. Guanylate cyclase B (particulate guanylate cyclase stimulated by C-type natriuretic peptide) and guanylate cyclase C (particulate guanylate cyclase stimulated by uroguanylin) activities were unaltered by pregnancy. The selective increase in responsiveness of particulate guanylate cyclase to atrial natriuretic peptide during pregnancy was not paralleled by an increased in level of messenger ribonucleic acid for the gene for guanylate cyclase A. CONCLUSION: Pregnancy reduced the in vitro responsiveness of the myometrial soluble guanylate cyclase to nitric oxide while increasing the responsiveness of the particulate isoform to atrial natriuretic peptide and brain natriuretic peptide through a mechanism independent of any change in receptor expression.

Effects of trepelennamine on brain monoamine turnover in morphine dependent and abstinent mice.

Previous studies have reported that the histamine H1 receptor blocker tripelennamine potentiates morphine withdrawal. In this paper, the in vivo effects produced by tripelennamine on the turnover of serotonin (5-HT), dopamine (DA) and noradrenaline (NA) in the whole brain, excluding the cerebellum, were studied in control, morphine-dependent (by SC implantation of a 75 mg morphine pellet) and morphine-dependent male CD1 mice just before naloxone-precipitated withdrawal. Tripelennamine (1-10 mg/kg) was administered SC 45 min. before the animals were killed. Serotonin, 5-hydroxyindole-3-acetic acid (5-HIAA), dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA) and noradrenaline were measured by high performance liquid chromatography coupled with electrochemical detection (HPLC-ECD) and 3-methoxy-4-hydroxyphenylethyleneglycol (MHPG) was measured by HPLC coupled with fluorimetric detection. Ratios 5-HIAA/ 5-HT, DOPAC + HVA/DA and MHPG/NA were taken as an index of serotonin, dopamine and noradrenaline turnovers, respectively. Tripelennamine (1 and 10 mg/kg) significantly reduced serotonin turnover in control and morphine-dependent mice, and potentiated the serotonin turnover reduction when it was administered 30 min before naloxone injection. The dopamine turnover was diminished by tripelennamine (1 and 10 mg/kg) in the morphine-dependent group. Tripelennamine (10 mg/kg) reduced noradrenaline turnover during abstinence. These results suggest that the potentiation of opiate abstinence by tripelennamine could be related to its antiserotonergic profile.

Morphine-induced changes in cerebral and cerebellar nitric oxide synthase activity.

The effect of acute and chronic morphine treatment on nitric oxide (NO) synthase activity (determined by the rate of conversion of [14C]arginine into [14C]citrulline) on mouse brain was studied. Acute morphine treatment induced an increased in Ca2+ -dependent NO synthase in cerebellum. This effect was blocked by coadministration with naloxone. Chronic morphine treatment (by s.c. pellet) also produced an increase in cerebellar NO synthase, with a maximum on the second day of implantation. No significant changes were found in frontal cortex and forebrain during acute or chronic morphine treatment. The relationship between opiate effects and the L-arginine: NO pathway is discussed.

Differential effects of chronic treatment with either dopamine D1 or D2 receptor agonists on the acute neuroendocrine actions of the highly potent synthetic cannabinoid HU-210 in male rats.

Acute exposure to delta 9-tetrahydrocannabinol (THC), the main psychoactive constituent of marijuana, produces a well-characterized set of neuroendocrine effects. The recent description of both brain cannabinoid receptors (CB-1) and anandamide, their proposed endogenous ligand, has renewed the interest in cannabinoid actions in the brain. However, the neurobiological mechanisms underlying the neuroendocrine effects of natural cannabinoids are not yet fully understood because of several mechanisms involved in their actions. In this work we have studied the role of hypothalamic dopaminergic receptors in the mediation of the acute neuroendocrine effects of (-)-delta 8-tetrahydrocannabinol-dimethyl-heptyl (HU-210, 20 micrograms/kg), a highly potent agonist of CB-1. The use of this low dose of HU-210 precludes the multiple unspecific effects which appear with an equipotent dose of THC. Rats were exposed during 21 days to either the dopamine (DA) D2 receptor agonist quinpirole (1 mg/kg, daily), the DA D1 receptor agonist SKF 38393 (8 mg/kg, twice a day) or vehicle (twice a day). Twenty-four hours after the last injection, a single dose of HU-210 (20 micrograms/kg) was administered intraperitoneally, and the animals were sacrificed 90 min later. Acute exposure to HU-210 produced both a decrease in plasma prolactin and a rise of plasma corticosterone levels. HU-210 treatment also resulted in both an increase in the L-3,4-dihydroxyphenylacetic acid/DA ratio and a decrease in noradrenaline contents, measured in the medial basal hypothalamus. These neuroendocrine actions were prevented by chronic exposure to quinpirole, but not after chronic SKF 38393 treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Changes induced by sodium cromoglycate in brain catecholamine turnover in morphine dependent and abstinent mice.

The effects of sodium cromoglycate (CRO) were studied in relation to the metabolism of brain catecholamines: dopamine (DA) and noradrenaline (NA), and their metabolites 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA) and 4-hydroxy-3-methoxyphenylethyleneglycol (MHPG). CRO was injected SC in control mice, morphine-tolerant mice (tolerance was induced by SC implantation of a 75 mg morphine pellet; CRO was administered on day 4 of addiction) and 30 min before abstinence (withdrawal was induced by SC injection of naloxone (1 mg/kg) on day 4 of addiction). Brain catecholamines and their metabolites were measured using high performance liquid chromatography coupled with electrochemical detection (HPLC-ECD), for DA, NA, DOPAC and HVA, and coupled with fluorescence detection for MHPG. The ratios of DOPAC + HVA/DA and MHPG/NA were kept as an index of DA and NA turnovers, respectively. CRO administered 30 min before naloxone-precipitated withdrawal diminished significantly NA levels in frontal cortex. CRO increased DA turnover in striatum and frontal cortex in naive animals and significantly diminished DA levels in frontal cortex and DOPAC levels in frontal cortex and midbrain in morphine-dependent mice. These findings are discussed in relation to the protective effects of CRO on opiate withdrawal and the effects of CRO on locomotor activity.

Changes induced by sodium cromoglycate on brain serotonin turnover in morphine dependent and abstinent mice.

This study was designed to explain the action of sodium cromoglycate (CRO) on the brain serotonergic system in control, morphine tolerant (by SC implantation of a 75 mg morphine pellet), and also in morphine dependent mice just before naloxone-precipitated withdrawal. After SC injections of CRO in control mice, morphine tolerant mice (day 4 of addiction), and 1 h before abstinence (withdrawal was induced by SC injection of 1 mg/kg naloxone on day 4 of addiction), animals were decapitated and various brain areas were rapidly removed. 5HT (Serotonin) and 5HIAA (5-hydroxyindole-3-acetic acid) were measured by high performance liquid chromatography coupled with electrochemical detection (HPLC-ECD). The ratio 5HIAA/5HT provided one index by which the turnover of the indoleamine was measured. CRO increased the turnover of 5HT in most of the brain areas studied in both control and morphine dependent mice. Furthermore, previous administration of CRO prior to naloxone challenge induced a significant increase in the 5HIAA/5HT ratio in the hypothalamus and striatum. These results are discussed as the reason for the preventive effect of CRO on jumping behaviour in morphine abstinent mice.

Role of sodium cromoglycate on analgesia, locomotor activity and opiate withdrawal in mice.

The role of sodium cromoglycate (CRO) on analgesia, locomotor activity and morphine withdrawal in mice was studied in morphine-dependent and drug-naive mice. CRO (0.5, 1, 5, 10, 30, 50 and 100 mg/kg, SC) induces analgesia (hot plate), an effect blocked by previous administration of the opiate antagonist naloxone (1 mg/kg). Furthermore, CRO (30 mg/kg) potentiates morphine analgesia. In morphine-tolerant mice, moderate doses of CRO (0.5, 1, 5, 10 and 30 mg/kg) do not induce analgesia, which suggested the development of cross tolerance between CRO and morphine, whereas coadministration of CRO and morphine in morphine tolerant animals restored the sensitivity to morphine. Administration of CRO (10 and 30 mg/kg) induces an increase in spontaneous locomotor activity, and previous administration of naloxone (1 mg/kg) blocks this effect, whereas CRO (10 mg/kg) blocks morphine (10 mg/kg) and amphetamine (3 mg/kg)-induced hyperactivity. CRO (10, 50 and 100 mg/kg) induces a significant and dose-dependent reduction in the number of jumps ("jumping up") during naloxone (1 mg/kg)-induced withdrawal in morphine-dependent mice. Finally, CRO (100 mg/kg) reduces the "wet dog shake" phenomenom during naloxone-induced withdrawal in morphine-dependent mice. These results suggest a possible stabilizing effect of CRO on the membranes of neurones that mediate analgesia, locomotor activity and opiate abstinence. Changes and inhibition of DA, NA and 5-HT release may also explain these effects.