Mitsuhiro Furuse

Inhibitory effect of ghrelin on food intake is mediated by the corticotropin-releasing factor system in neonatal chicks

It is known that, in rats, central and peripheral ghrelin increases food intake mainly through activation of neuropeptide Y (NPY) neurons. In contrast, intracerebroventricular (ICV) injection of ghrelin inhibits food intake in neonatal chicks. We examined the mechanism governing this inhibitory effect in chicks. The ICV injection of ghrelin or corticotropin-releasing factor (CRF), which also inhibits feeding and causes hyperactivity in chicks. Thus, we examined the interaction of ghrelin with CRF and the hypothalamo-pituitary-adrenal (HPA) axis. The ICV injection of ghrelin increased plasma corticosterone levels in a dose-dependent or a time-dependent manner. Co-injection of a CRF receptor antagonist, astressin, attenuated ghrelin-induced plasma corticosterone increase and anorexia. In addition, we also investigated the effect of ghrelin on NPY-induced food intake and on expression of hypothalamic NPY mRNA. Co-injection of ghrelin with NPY inhibited NPY-induced increase in food intake, and the ICV injection of ghrelin did not change NPY mRNA expression. These results indicate that central ghrelin does not interact with NPY as seen in rodents, but instead inhibits food intake by interacting with the endogenous CRF and its receptor.

Intracerebroventricular injection of orexins does not stimulate food intake in neonatal chicks

1. Recently, 2 novel neuropeptides were discovered, both derived from the same precursor by proteolytic processing, which bind and activate 2 closely related orphan G protein-coupled receptors, Named orexin-A and -B (Sakurai et al., 1998). Both stimulate food intake when administered centrally to rats. 2. Our aim was to elucidate whether central injection of mammalian orexin-A or -B stimulates food intake in the chick. 3. Under conditions of free access to food, orexin-A did not alter the food intake of chicks, but cumulative food intake was significantly suppressed by orexin-B. 4. The orexin-B was then administered to chicks deprived of food for 3 h to confirm its suppressive effect. No significant effect of orexin-B on food intake was detected. 5. Central injection of orexin-B did not modify food intake when appetite was stimulated by fasting. 6. Neither of these orexins appears to stimulate feeding in chicks.

Intracerebroventricular injection of ghrelin and growth hormone releasing factor inhibits food intake in neonatal chicks

Growth hormone releasing factor (GRF) is known to stimulate feeding of rats. Ghrelin, a novel growth hormone (GH)-releasing acylated peptide, was recently isolated from rat stomach. It also stimulates the release of GH from the anterior pituitary through the GH secretagogue receptor (GHS-R) and feeding in the rat. We have investigated the effects of ghrelin and GRF on food intake of the neonatal chick. In Experiment 1, 0, 1.25, 2.5 and 5 microg of ghrelin were administered intracerebroventricularly (i.c.v.) to ad libitum fed birds. In Experiment 2, the effect of (i.c.v.) injection of 0, 1.25, 2.5 and 5 microg of GRF was investigated. Both peptides strongly inhibited food intake of the chick during the 2-h post-injection period. In the third experiment, 0, 0.5, 1 and 2 microg of ghrelin was injected i.c.v. in chicks previously deprived of food for 3 h. Food intake was again inhibited by ghrelin in a dose-dependent manner. These results suggest that the mechanisms for feeding of the neonatal chick through GH release are different from mammals.

The central corticotropin-releasing factor and glucagon-like peptide-1 in food intake of the neonatal chick

Recently, we have reported that central administration of glucagon-like peptide-1 (GLP-1) strongly decreases food intake of chicks. The aim of this study was to elucidate whether suppressed food intake induced by the central injection of GLP-1 is mediated by activation of the hypothalamic-pituitary-adrenal axis. First, the effects of central administration of corticotropin-releasing factor (CRF) were investigated. Birds (2-day-old) were food-deprived for 3 h and then CRF or saline was injected intracerebroventricular (i.c.v.). CRF strongly inhibited food intake. Thereafter, effects of central CRF or GLP-1 on plasma corticosterone concentration were examined. CRF significantly stimulated corticosterone release, but GLP-1 did not alter plasma corticosterone concentration. These results suggest that CRF is a potent inhibitor of food intake in the chick, but the suppression of food intake induced by central GLP-1 may not be involved in the activation of hypothalamic-pituitary-adrenal axis.

Commensal microbiota modulate murine behaviors in a strictly contamination-free environment confirmed by culture-based methods

There is increasing evidence suggesting the existence of an interaction between commensal microbiota, the gut and the brain. The aim of this study was to examine the influence of commensal microbiota on the host behaviors in a contamination‐free environment, which was verified by culture‐based methods.

Gonadotropin-inhibiting hormone stimulates feeding behavior in chicks.

Neuropeptides containing a C-terminal Arg-Phe-NH2 motif (RFamide peptides) are suggested to be involved in the control of feeding behavior in both invertebrates and vertebrates. Gonadotropin-inhibitory hormone (GnIH) is the first identified avian RFamide peptide that inhibits gonadotropin release from the pituitary. The GnIH precursor encodes one GnIH and its related peptides (GnIH-RP-1 and -RP-2) that shared the same C-terminal motif, Leu-Pro-Xaa-Arg-Phe-NH2 (Xaa = Leu or Gln) (LPXRFamide). GnIH neurons are localized in the paraventricular nucleus, with their fibers visible in multiple brain locations including the median eminence and brainstem. In this study, we therefore investigated the action of GnIH and its related peptides on feeding behavior. Intracerebroventricular (ICV) injection of GnIH, GnIH-RP-1 and GnIH-RP-2 significantly stimulated food intake in chicks. The chicken pentapeptide LPLRFamide, a degraded C-terminus of GnIH and GnIH-RP-1, did not stimulate feeding thereby demonstrating the importance of the N-terminus of GnIH and its related peptides for the orexigenic effect. Anti-GnIH antiserum suppressed appetite induced by fasting, but did not modify feeding under ad libitum conditions. The present study suggests that GnIH and its related peptides act as endogenous orexigenic factors in the brain of chicks.

Chicken ghrelin and growth hormone-releasing peptide-2 inhibit food intake of neonatal chicks.

Ghrelin is an endogenous ligand for the growth hormone secretagogue (GHS) receptor. Ghrelin stimulates feeding in rats, however, intracerebroventricular (i.c.v.) injection of rat ghrelin inhibits feeding of neonatal chicks. In the present study, the effect of i.c.v. injection of different ghrelins including chicken and bullfrog ghrelin, and synthetic GH-releasing peptide (GHRP) on feeding of neonatal chicks was investigated. Chicken ghrelin strongly suppressed feeding. To compare the inhibitory effect, chicken and rat ghrelin were examined. The suppressive effect of feeding by chicken and rat ghrelin was almost identical. Bullfrog ghrelin contains a change in the acylated amino acid from Ser to Thr, strongly suppressed feeding. The i.c.v. injection of GHRP-2 (KP-102), a synthetic GHS, also inhibited feeding. These results indicate that the chicken GHS receptor is affected by several forms of GHS, and that food intake of neonatal chicks is inhibited by GHS receptor agonists.

High concentrations of HGF inhibit skeletal muscle satellite cell proliferation in vitro by inducing expression of myostatin: a possible mechanism for reestablishing satellite cell quiescence in vivo

Skeletal muscle regeneration and work-induced hypertrophy rely on molecular events responsible for activation and quiescence of resident myogenic stem cells, satellite cells. Recent studies demonstrated that hepatocyte growth factor (HGF) triggers activation and entry into the cell cycle in response to mechanical perturbation, and that subsequent expression of myostatin may signal a return to cell quiescence. However, mechanisms responsible for coordinating expression of myostatin after an appropriate time lag following activation and proliferation are not clear. Here we address the possible role of HGF in quiescence through its concentration-dependent negative-feedback mechanism following satellite cell activation and proliferation. When activated/proliferating satellite cell cultures were treated for 24 h beginning 48-h postplating with 10-500 ng/ml HGF, the percentage of bromodeoxyuridine-incorporating cells decreased down to a baseline level comparable to 24-h control cultures in a HGF dose-dependent manner. The high level HGF treatment did not impair the cell viability and differentiation levels, and cells could be reactivated by lowering HGF concentrations to 2.5 ng/ml, a concentration that has been shown to optimally stimulate activation of satellite cells in culture. Coaddition of antimyostatin neutralizing antibody could prevent deactivation and abolish upregulation of cyclin-dependent kinase (Cdk) inhibitor p21. Myostatin mRNA expression was upregulated with high concentrations of HGF, as demonstrated by RT-PCR, and enhanced myostatin protein expression and secretion were revealed by Western blots of the cell lysates and conditioned media. These results indicate that HGF could induce satellite cell quiescence by stimulating myostatin expression. The HGF concentration required (over 10-50 ng/ml), however, is much higher than that for activation, which is initiated by rapid release of HGF from its extracellular association. Considering that HGF is produced by satellite cells and spleen and liver cells in response to muscle damage, local concentrations of HGF bathing satellite cells may reach a threshold sufficient to induce myostatin expression. This time lag may delay action of the quiescence signaling program in proliferating satellite cells during initial phases of muscle regeneration followed by induction of quiescence in a subset of cells during later phases.

Intracerebroventricular injection of agouti-related protein attenuates the anorexigenic effect of alpha-melanocyte stimulating hormone in neonatal chicks

It is well known that alpha-melanocyte stimulating hormone (alpha-MSH) inhibits feeding via melanocortin receptor-4 (MC4R) in the mammalian brain. The anorexigenic effect of alpha-MSH is attenuated by agouti-related protein (AGRP), an antagonist for MC4R. Present studies were carried out to clarify whether human AGRP (86-132) antagonizes the anorexigenic effect of alpha-MSH in broiler chicks. Intracerebroventricular injection of AGRP attenuated the anorexigenic effect of alpha-MSH. Furthermore, AGRP stimulated food intake of layer-type chicks under an ad libitum feeding condition but not broiler chicks, suggesting that the orexigenic effect of AGRP is different between two breeds. These also imply that the extent of the anorexigenic effect of endogenous alpha-MSH is different among two breeds. This may be a part of the difference in food intake between two breeds.

Inhibition of Food Intake by CRF in Chickens

The effect of intracerebroventricular (i.c.v.) injection of corticotrophin releasing factor (CRF) on food and water intake and on body temperature in chickens was determined. Both broiler and Leghorn type chickens were utilized in this experiment. A stainless steel guide cannula was surgically implanted into the right lateral ventricle of each bird. The i.c.v. injection of CRF significantly decreased food intake in both fed and overnight-fasted broilers and Leghorns. Water intake was decreased by CRF in Leghorns but not broilers. When CRF was injected into Leghorns given access to water, but not food, water intake was not affected. Body temperature was not affected by the i.c.v. injection of CRF. These results suggest that CRF acts within the central nervous system of chickens to decrease food intake while having no affect on water intake or body temperature.