Possible involvement of the dopamine D2 receptors of ventromedial hypothalamus in the control of free- and scheduled-feeding and plasma ghrelin level in rat.

OBJECTIVES: We investigated effect of the ventromedial hypothalamus (VMH) dopamine D2 receptor inhibition on food intake and plasma ghrelin following chronic free or scheduled meal with different caloric intakes. METHODS: Male Wistar rats (220-250 g) were fed diets containing free (control) or three scheduled diets of standard, restricted and high-fat for 1 month. The animals stereotaxically received an intra VMH single dose of sulpiride (0.005 µg)/or saline (0.5 µL) before meal time. Thirty minutes later, food intake and circulating ghrelin were measured. RESULTS: Sulpiride significantly reduced food intake and ghrelin concentration in freely fed and scheduled-standard diet (p<0.05), while increased food intake, with ghrelin level on fasted level in scheduled-restricted group (p<0.01) compared to control. Food intake and ghrelin concentration between scheduled-high fat and freely fed or scheduled-standard diets did not show significant changes. CONCLUSIONS: The VMH D2 receptors are possibly involved in controlling scheduled eating behavior, depending on energy balance context.

[The effect of the dopamine D2 receptor agonist cabergoline on the content of catecholamines and expression of BDNF mRNA in the rat midbrain and hypothalamus]. / Vliianie agonista dofaminovykh D2-retseptorov kabergolina na soderzhanie katekholaminov i ékspressiiu mRNK BDNF v srednem mozge i gipotalamuse v éksperimente.

AIM: To study an effect of cabergoline on dopamine and noradrenaline concentration and BDNF mRNA level in the rat midbrain and hypothalamus. MATERIAL AND METHODS: Twenty adult male Wistar rats were used in a single treatment paradigm: animals of the treatment group (n=10) received cabergoline (i.p., 0.5 mg/kg) and the control group (n=10) received an equivalent volume of the solvent. Quantitative analysis for the dopamine (DA) and noradrenaline (NA) was carried out using high-performance liquid chromatography (HPLC) coupled with electrochemical detection. BDNF mRNA levels were studied using quantitative RT-PCR. RESULTS AND CONCLUSION: Cabergoline significantly increases NA concentration in the midbrain 24 hours after injection: 639.2±64.5 ng/g in the treatment group versus 398.0±66.0 ng/g in the control group (p<0.05), while mean content of DA is not significantly changed (211.4±16.3 ng/g vs 169.7±54.6 ng/g, respectively). Cabergoline does not affect hypothalamic DA and NA levels. The drug increases BDNF mRNA levels by 2-times in the midbrain, but not in the hypothalamus, 24 hours after injection.

Guibourtia tessmannii-induced fictive ejaculation in spinal male rat: involvement of D1, D2-like receptors.

CONTEXT: Guibourtia tessmannii (Caesalpiniaceae) is a plant traditionally used as aphrodisiac. We previously reported the pro-ejaculatory effects of the aqueous and methanol extracts of G. tesmannii in spinal male rat. However, the mechanism underlying such effects has not been elucidated. OBJECTIVE: This study characterizes the dopaminergic sub-type receptors involved in G. tesmannii-induced ejaculation in male Wistar rat. MATERIALS AND METHODS: Urethane-anesthetized spinal male rats were intravenously treated with saline solution (1 mL/kg, control); dopamine (0.1 µmol/kg, reference); aqueous or methanol extracts of G. tesmannii (20 mg/kg) in the absence or presence of haloperidol (0.26 µmol/kg), a nonspecific dopaminergic receptor antagonist, Sch23390 (0.26 µmol/kg), a specific D1-like receptor antagonist or, sulpiride (0.26 µmol/kg), a specific D2-like receptor antagonist. Electromyography of the bulbospongiosus muscles and intraseminal pressure were recorded after urethral, penile and drug stimulations. RESULTS: Urethral and penile stimulations, intravenous injection of dopamine or, aqueous and methanol extracts of G. tesmannii always triggered the expression of rhythmic contraction of the bulbospongiosus muscles with an average mean of 3.33 ± 0.43; 7.83 ± 0.85; 9.80 ± 0.86; 0.83 ± 0.54 and 2.67 ± 0.95 contractions, respectively. The intraseminal pressure was more expressed after urethral and penile stimulations (15.66 ± 1.58 and 13.60 ± 2.40 mmHg, respectively). In rats pretreated with haloperidol, Sch23390 or sulpiride, no ejaculation was recorded after intravenous injection of G. tesmannii extracts or dopamine. DISCUSSION AND CONCLUSION: Guibourtia tesmannii-induced ejaculation requires the integrity of D1 and D2-like receptors. These findings further justify the ethno-medicinal claims of G. tesmannii as an aphrodisiac.

Use of radiolabeled antagonist assays for assessing agonism at D2 and D3 dopamine receptors: comparison with functional GTPγS assays.

BACKGROUND: Cell-based drug screening assays are essential tools for drug discovery and development targeting G protein-coupled receptors, which include dopamine D3 receptors. D3 is notorious for its poor coupling to G protein in most heterologous cell lines, and therefore D3 agonist-stimulated binding of [(35)S]GTPγS to G protein cannot be observed in many "non-functional" D3 expressing cell lines. NEW METHOD: The present work explores the use of an alternate method for assessing agonist activity, consisting of measuring the difference in agonist competition between [(3)H]spiperone bound to low-affinity states of the receptor and that with radioligand bound to high-affinity states (GTP shift assay). COMPARISON WITH EXISTING METHOD: The current study describes the determination of GTP shifts in [(3)H]spiperone binding assays for the assessment of agonists' potencies (at D2 and D3) and efficacies (at D3). Compared with GTPγ(35)S binding assays, the new method removes the cumbersome need of functional D3 cell lines and limited project duration due to short half-life of isotope (35)S. CONCLUSION: The new method allows the estimation of potency (D2 and D3) and efficacy (D3) at the level of receptor and G protein activation in a simple fashion from shifts in monophasic-inhibition curves. Moreover, it does not require [(35)S]GTPγS binding assays with functional D3 cells. This method will have wide applicability for D3-selective agonist screening. It may also be useful for other GPCRs circumventing the need for functional assays and offering the ability to detect agonist activity regardless of the particular signaling pathway.