Stimulation Of D2-like Receptors Research Articles

Conditioned Grooming Induced by the Dopamine D1-like Receptor Agonist SKF 38393 in Rats

Stimulation of dopamine D1-like receptors reliably increases grooming in rats and mice. The study examined whether the grooming response elicited by the prototypical D1-like agonist SKF 38393 (8 mg/kg SC) could be conditioned to the specific environment in which it occurred. Rats in one group (Paired) received SKF 38393 and rats in another group (Unpaired) received saline in observation boxes outside of their housing room; the rats were then scored for duration and frequency of grooming bouts over 25 min. The ordering of injections was reversed the next day in the rats, housing room. The procedure was repeated twice, with at least one intervening drug-free day, to give three conditioning trials. The D1-like agonist significantly increased grooming on each of the three conditioning trials, without obvious tolerance or sensitization, and the effect tended to persist for the duration of each trial. On the test trial for conditioned grooming, mean grooming duration was significantly greater in the Paired than the Unpaired group, suggesting that SKF 38393-induced grooming had been conditioned to the test environment. This is the first time that drug-elicited grooming has been conditioned to environmental cues.

Activation of type II adenylate cyclase by D2 and D4 but not D3 dopamine receptors.

The D2-like dopamine receptors couple to a variety of signal transduction pathways, including inhibition of adenylate cyclase, mitogenesis, and activation of potassium channels. Although these effects are mediated via pertussis toxin-sensitive G proteins, G(i/o), it is likely that some of these effects are influenced by the release of G protein betagamma subunits. Type II adenylate cyclase (ACII) is highly regulated by multiple biochemical stimuli, including protein kinase C, forskolin, G protein alpha subunits, and G protein betagamma subunits. The ability of betagamma subunits to activate this enzyme in the presence of activated alpha(s) has been particularly well characterized. Although stimulation by betagamma subunits has been described as conditional on the presence of activated alpha(s), betagamma subunits also potentiate ACII activity after activation of protein kinase C. We created stable cell lines expressing ACII and the D2L receptor, the D3 receptor, or the D4.4 receptor. Activation of D2L or D4.4 receptors, but not D3 receptors, potentiated beta-adrenergic receptor/Gs-stimulated activity of ACII, as measured by the intracellular accumulation of cAMP. Similarly, stimulation of D2L or D4.4 receptors potentiated phorbol ester-stimulated ACII activity in the absence of activated alpha(s), whereas stimulation of D3 receptors did not. The effect of D2-like receptor stimulation was blocked by pretreatment with pertussis toxin and by inhibition of protein kinase C. We propose that activation of both D2L and D4.4 dopamine receptors potentiated phorbol-12-myristate-13-acetate-stimulated ACII activity through the release of betagamma subunits from pertussis toxin-sensitive G proteins. In contrast, the lack of D3 receptor-mediated effects suggests that stimulation of D3 receptors does not result in an appreciable release of betagamma subunits.

Dopamine D1-like receptor stimulation inhibits hypertrophy induced by platelet-derived growth factor in cultured rat renal vascular smooth muscle cells.

Vascular smooth muscle cell (VSMC) hypertrophy is believed to play some roles in atherosclerosis. To elucidate the role of vascular D1-like receptors in VSMC hypertrophy, the effects of dopamine and specific D1-like receptor agonists SKF 38393 and YM 435 on platelet-derived growth factor (PDGF) BB-mediated VSMC hypertrophy was studied. We observed that cells stimulated by PDGF-BB 5 ng/mL showed increased VSMC hypertrophy. These effects were prevented by coincubation with dopamine, SKF 38393, and YM 435 1-10 mumol/L, and this prevention was reversed by Sch 23390 1 to 10 mumol/L, a specific D1-like receptor antagonist. These actions are mimicked by forskolin 1 to 10 mumol/L, a direct activator of adenylate cyclase and 8-bromo-cAMP 0.1 to 1 mmol/L, and are blocked by a specific protein kinase A (PKA) inhibitor N-[2-(P-bromcoinnamylamino)ethyl]-5-isoquinoline-sulfonamide (H89) but not blocked by its negative control. PDGF-BB (5 ng/mL)-mediated mitogen-activated protein kinase (MAPK) activity was significantly suppressed by coincubation with D1-like receptor agonists, which were reversed by PKA inhibitor H 89. These results suggest that vascular D1-like receptor agonists inhibit hypertrophy of VSMC, possibly through PKA activation and suppression of activated MAPK activity.

Role of D1 and alpha1 receptors in the enhanced locomotor response to dopamine D2-like receptor stimulation induced by repeated electroconvulsive shock.

We previously reported that, in rats chronically treated with the antidepressant drug imipramine, the enhanced locomotor response to the D2-like receptor agonist quinpirole became less sensitive to the inhibitory effect of the D1 receptor antagonist SCH 23390 and more sensitive to the inhibitory effect of the alpha1 receptor antagonist prazosin. In this study, we show that in electroconvulsive shock-treated rats these antagonists behave in the opposite manner to that observed in imipramine-treated rats, with SCH 23390 being highly effective and prazosin ineffective in antagonizing the locomotor response to quinpirole. The possibility that these differences may reflect some of the clinical characteristics of these antidepressant treatments is discussed.

The effects of the addition of low-dose dopamine on renal and cardiac function in septic critically-ill patients: A dose finding study

Dopamine acts as an inodilator with diuretic properties at the 2μg/kg/min dose. The diuretic properties are mainly due to tubular D1-like receptor stimulation since the creatinine clearance was not improved by dopamine. Higher doses of dopamine (4 and 6 μg/kg/min) further increase diuresis and sodium excretion, but this is accompanied by an increase of heart rate. Whether the enhancement of diuresis in septic patients, who in general need ongoing filling, is desirable remains to be answered.

Induction of c-fos and zif/268 gene expression in rat striatal neurons, following stimulation of D1-like dopamine receptors, involves protein kinase A and protein kinase C.

Changes in the level of dopaminergic activity in the rat striatum lead to the induction of a number of immediate-early genes, including c-fos and zif/268. These immediate-early genes are thought in turn to alter the rate of transcription of downstream genes. There is evidence that the dopaminergic activation of the c-fos and zif/268 genes in the striatum in vivo is linked to stimulation of D1-like dopamine receptors. We have used primary cultures of embryonic rat striatal neurons to identify the intracellular pathways involved in this response. Dopamine (10 nM-5 microM) caused a marked increase in the levels of c-fos mRNA and zif/268 mRNA in cultured striatal neurons, an effect that was reproduced by the D1-like dopamine receptor agonist SKF38393 (10 nM-5 microM). These actions were attenuated by the D1-like antagonist SCH23390 (1 microM) but not by the D2-like antagonist eticlopride (1 microM). The D2-like agonist quinpirole did not increase zif/268 mRNA above basal levels at concentrations up to 5 microM, but caused a slight increase in the levels of c-fos mRNA. The stimulation of c-fos mRNA levels caused by 1 microM SKF38393 was reduced by 45% following pretreatment with the selective protein kinase A inhibitor KT5720, and by 87% following pretreatment with the selective protein kinase C inhibitor calphostin C. The stimulation of zif/268 mRNA levels caused by 1 microM SKF38393 was reduced by 90% following pretreatment with KT5720, but was not significantly affected by pretreatment with calphostin C. In addition, the actions of SKF38393 to stimulate the expression of both immediate-early genes were attenuated by coadministration of quinpirole. These results suggest that SKF38393 acts on striatal neurons to stimulate c-fos expression predominantly through protein kinase C, but also partially through protein kinase A. Conversely, SKF38393 induces zif/268 expression through protein kinase A. The ability of quinpirole to antagonize the actions of SKF38393 on cultured neurons is consistent with the presence of both D1-like receptors on the same neuronal population.

Anatomical localization of SKF-38393-induced behaviors in rats using the irreversible monoamine receptor antagonist EEDQ.

This study was designed to localize the population of dopamine D1-like receptors involved in grooming and oral movements elicited by systemic administration of the D1-selective agonist SKF-38393. Receptors in specific dopamine terminal regions were inactivated by intracranial injection of the nonselective irreversible antagonist N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ). The effect of these injections on behaviors induced by systemic administration of SKF-38393 (10 mg/kg) was measured 48 hours later. The specific populations of D1-like receptors inactivated by the EEDQ injections were identified as a loss of 3H-SCH-23390 binding in a given region using quantitative autoradiography. EEDQ (1.5 micrograms/microliters/side) injected into the nucleus accumbens (NAc) did not alter SKF-38393-induced behaviors. Similarly, injection of EEDQ into the medial caudate-putamen (CPu) failed to alter these behaviors. In contrast, EEDQ (0.15-1.5 micrograms/microliters/side) injected into the lateral CPu decreased both SKF-38393-induced grooming and oral movements, with complete blockade of grooming observed at the highest dose. To determine whether this effect of EEDQ was due to inactivation of D1-like receptors, separate groups of animals were pretreated with SCH-23390 (3 mg/kg, S.C.) 15 min prior to injection with EEDQ. Pretreatment with SCH-23390 prevented the disruption of SKF-38393-induced behaviors, as well as the loss of 3H-SCH-23390-labeled binding sites observed after injection of EEDQ into the lateral CPu. EEDQ injections that produced disruption of SKF-38393-induced behaviors were associated with a greater loss of binding in the lateral CPu relative to other regions examined including the NAc, medial CPu, and globus pallidus. Furthermore, EEDQ injections that produced the greatest loss of 3H-SCH-23390 binding in the latter three regions did not disrupt SKF-38393-induced behavior. These results demonstrate that stimulation of D1-like receptors in the lateral CPu is necessary for behaviors induced by systemic administration of SKF-38393. The results also demonstrate the utility of this "receptor lesion" technique to localize receptor-mediated behaviors.

Dopamine and glutamate agonists stimulate neuron-specific expression of Fos-like protein in the striatum.

1. The monoamine dopamine and the amino acid glutamate are major neurotransmitters in the basal ganglia implicated in the normal functions of the striatum and in extrapyramidal disease states. To study the effects of these neurotransmitters on gene transcription in striatal neurons, we treated rats with dopamine (monoamine) agonists and with glutamate agonists and monitored the induction of Fos-like protein in striatal neurons. We administered the indirect monoamine agonists cocaine and amphetamine intraperitoneally and gave the glutamate agonist quinolinic acid by direct intrastriatal injection. We identified the phenotypes of the responsive neurons by immunohistochemistry and by enzyme histochemistry in double staining protocols. 2. Both the indirect monoamine agonists and the glutamate receptor agonist stimulated rapid nuclear expression of Fos-like protein in specific classes of striatal neurons. The induction by cocaine and amphetamine was blocked by pretreatment with the dopamine D1-like receptor antagonist SCH23390, and the induction by quinolinic acid was blocked by pretreatment with MK-801, a noncompetitive antagonist of the N-methyl-D-aspartate (NMDA) glutamate receptor. 3. The monoamine and glutamate agonists both induced Fos-like protein exclusively in striatal neurons that constitutively expressed the protein phosphatase inhibitor DARPP-32 (dopamine and cAMP-regulated phosphoprotein). 4. The dopamine agonists failed to induce detectable Fos-like protein in striatal neurons expressing enkephalin, even though many such neurons expressed DARPP-32. By contrast, many enkephalinergic neurons did express Fos-like protein in response to glutamatergic stimulation. 5. Glutamate agonist stimulation, but not dopamine agonist stimulation, induced Fos-like protein in a subpopulation of striatal interneurons, namely, a group of neurons exhibiting NADPH-diaphorase activity. 6. These findings suggest that stimulation of dopamine D1-like receptors (or related monoamine receptors) and glutamate NMDA receptors activates neuron-specific programs of immediate-early gene expression in the striatum. Our findings further suggest that monoamine and glutamate may act cooperatively at the transcriptional level on a functionally defined subset of striatal neurons.