Amphetamine-regulated Transcript Expression Research Articles

Regulation of Pituitary Cocaine- and Amphetamine-Regulated Transcript Expression and Secretion by Hypothalamic Gonadotropin-Releasing Hormone in Chickens.

There is increasing evidence that cocaine- and amphetamine-regulated transcript (CART) peptide is abundantly expressed in the anterior pituitary of birds and mammals, suggesting that CART peptide may be a novel pituitary hormone and its expression and secretion is likely controlled by the hypothalamic factor(s). To substantiate this hypothesis, using chicken as an animal model, we examined the effects of gonadotropin-releasing hormone (GnRH) on pituitary CART secretion and expression and investigated whether GnRH could modulate plasma CART levels. The results showed that: (1) chicken GnRH (GnRH1 and GnRH2) could potently stimulate CART peptide secretion in intact pituitaries incubated in vitro, as detected by Western blot; (2) GnRH could also stimulate CART mRNA expression in cultured pituitary cells, as revealed by quantitative real-time polymerase chain reaction (qPCR) assay; (3) GnRH actions on pituitary CART expression and secretion are likely mediated by GnRH receptor coupled to the intracellular Ca2+, MEK/ERK, and cAMP/PKA signaling pathways; and (4) plasma CART levels are high in chickens at various developmental stages (1.2–3.5 ng/ml) and show an increasing trend towards sexual maturity, as detected by enzyme-linked immunosorbent assay (ELISA). Moreover, plasma CART levels could be significantly induced by intraperitoneal administration of GnRH in chicks. Taken together, our data provide the first collective evidence that CART peptide is a novel pituitary hormone and its expression and secretion are tightly controlled by hypothalamic GnRH, thus likely being an active player in the hypothalamic-pituitary-gonadal (HPG) axis.

Passiflora edulis peel intake improves insulin sensitivity, increasing incretins and hypothalamic satietogenic neuropeptide in rats on a high-fat diet

ObjectiveThis study aimed to investigate the effect of Passiflora edulis peel flour (PEPF) intake on hypothalamic neuropeptides messenger RNA expression, insulin sensitivity, and other metabolic parameters in Sprague-Dawley rats fed a high-fat (HF) diet. MethodsSprague-Dawley rats were divided in 3 groups: a control group, fed on a normal fat diet; a HF group, fed on a high-fat diet (35% fat [w/w]); and a high-fat Passiflora flour (HFPF) group, fed on a HF diet containing PEPF. The rats from the HFPF group as well as the HF group were kept on an HF diet for the first 4 wk to induce metabolic conditions related to obesity. Then the HFPF group was switched to a HF diet containing PEPF for additional 6 wk. Other groups were kept on normal-fat and HF diet without addition of PEPF during the whole period of experiment. The glucose tolerance and insulin sensitivity were evaluated through the glucose tolerance test (GTT) and the insulin tolerance test (ITT). Gut hormones and adipokines were measured through an immunoassay. The hypothalamic neuropeptides expression was assessed by real-time polymerase chain reaction. ResultsThe PEPF intake increased the hypothalamic cocaine- and amphetamine-regulated transcript expression (CART) (P < 0.05), counteracted cumulative body weight gain (P < 0.001), decreased adiposity (P < 0.05) and leptin level (P < 0.01), whereas increased adiponectin (P < 0.01), glucose-dependent insulinotropic polypeptide (P < 0.01), and glucagon-like peptide-1 (GLP-1) (P < 0.001) improved the insulin sensitivity in diet-induced obesity rats by increasing the kITT (glucose disappearance rate) (P < 0.01), which was calculated during the ITT. Other gut hormones (peptide tyrosine tyrosine, pancreatic polypeptide, and amylin) and interleukins (IL) (IL-6, tumor necrosis factor-α, IL-1β, and monocyte chemoattractant protein-1) were not changed by the PEPF intake. ConclusionOur findings provide a further understanding of how the PEPF works as a dietary component to improve glucose homeostasis and demonstrate a molecular mechanism that may increase satiety by PEPF in diet-induced obesity.

Time course effects of adrenalectomy and food intake on cocaine- and amphetamine-regulated transcript expression in the hypothalamus

Cocaine- and amphetamine-regulated transcript (CART) has been implicated in the feeding behavior and the regulation of hypothalamic–pituitary–adrenal axis activity. In this study we investigated the expression of CART mRNA in the hypothalamus at several intervals after adrenalectomy or sham surgery in basal conditions or after a fasting–refeeding regimen. Male Wistar rats, with free access to food and drinking, were subjected to bilateral adrenalectomy (ADX) or sham surgery. Plasma corticosterone, ACTH, and leptin levels, epididymal and perirenal fat content, and CART expression were determined 1, 3, 7 and 14 days after surgery. Another set of rats was subjected to a 48-h fasting period followed by refeeding during 4 h on the 7th day after ADX or sham surgery. On the day of the experiment, rats were anesthetized and perfused and the brain was processed for CART mRNA in situ hybridization. We observed that long-term but not short-term adrenalectomy decreased leptin plasma levels and CART expression in the arcuate and paraventricular nuclei. Furthermore, we showed that CART expression was reduced by fasting and it was increased after refeeding in the sham group, however, CART expression was not changed by fasting or refeeding after ADX. In conclusion, the present data indicate that following long-term ADX, under freely feeding conditions, there is a decrease of CART expression in the hypothalamus that is associated with a decrease of leptin secretion. CART expression induced by feeding seems to be modulated by glucocorticoid.

Clozapine and dopamine D3 receptor antisense reduce cocaine- and amphetamine-regulated transcript expression in the rat nucleus accumbens shell.

Cocaine- and amphetamine-regulated transcript (CART) is a neuropeptide highly expressed in the hypothalamus and nucleus accumbens. Hypothalamic CART has been associated with food intake and body weight control, but in the nucleus accumbens the role of CART remains elusive. New generations of antipsychotic drugs show a good efficacy over psychotic symptoms but they induce an important weight gain. The mechanism underlying this unexpected side effect is still unknown. The present results show, for the first time, that acute and chronic treatment with the atypical neuroleptic clozapine, but not the conventional neuroleptic haloperidol, reduced the expression of CART mRNA in the shell of the nucleus accumbens. CART mRNA is colocalized with both dopamine D(2) and D(3) receptor transcripts in the nucleus accumbens shell. However, a dopamine D(3) receptor, but not D(2), antisense oligonucleotide administration reduced CART expression in this brain area. These results suggest that the dopamine D(3) receptor, but not the D(2), is involved in the control of CART expression in the nucleus accumbens and that it may participate in the modulation of CART mRNA levels by clozapine. The modulation of CART, an anorexigenic neuropeptide, in the dopamine mesolimbic pathway may potentially play a role in dysregulated food intake induced by some antipsychotic drugs.

Thyroid status regulates CART but not AgRP mRNA levels in the rat hypothalamus.

We examined the effects of thyroid status on cocaine- and amphetamine-regulated transcript and agouti-related peptide expression in the rat hypothalamus. Hypo- and hyperthyroidism were induced in adult male rats, and the mRNA content of cocaine- and amphetamine-regulated transcript and agouti-related peptide was determined using in situ hybridization. Hyperthyroidism induces a reduction in cocaine- and amphetamine-regulated transcript mRNA levels in the paraventricular nucleus, without any change in the arcuate and dorsomedial nuclei and in the lateral hypothalamic area. On the other hand, hypothyroidism had not effect on cocaine- and amphetamine-regulated transcript expression in any of these nuclei. Agouti-related peptide expression in the arcuate nucleus was not affected by the thyroid status. These data indicate that the increments in food intake in hyperthyroidism could be mediated, at least in some extent, by a decreased expression, at the paraventricular nucleus of the hypothalamus, of the anorexigenic cocaine- and amphetamine-regulated transcript peptides.

Hyperleptinemia in A(y)/a mice upregulates arcuate cocaine- and amphetamine-regulated transcript expression.

The effects of leptin on cocaine- and amphetamine-regulated transcript (CART) and agouti-related protein (AGRP) expression in the hypothalamic arcuate nucleus of obese A(y)/a mice were investigated. CART mRNA expression was upregulated by 41% and AGRP mRNA downregulated by 78% in hyperleptinemic A(y)/a mice relative to levels in lean a/a mice. The mRNA expression of these neuropeptides in either young nonobese A(y)/a mice or rats treated with SHU-9119, a synthetic melanocortin-4 receptor (MC4R) antagonist, did not differ significantly from that in the corresponding controls. After a 72-h fast, which decreased the concentration of serum leptin, CART and AGRP mRNA expression decreased and increased, respectively, in A(y)/a mice. The expression levels of these neuropeptides in leptin-deficient A(y)/a ob/ob double mutants were comparable to those in a/a ob/ob mice. Leptin thus modulates both CART and AGRP mRNA expression in obese A(y)/a mice, whereas leptin signals are blocked at the MCR4R level. Taken together, the present findings indicate that differential expression of these neuropeptides in A(y)/a and ob/ob mice results in dissimilar progression toward obesity.