The role of nutritional profile in the orexigenic neuropeptide secretion in nonalcoholic fatty liver disease obese adolescents

Background Little progress has been made to identify the central neuroendocrine pathway involved in the energy intake control in nonalcoholic fatty liver disease (NAFLD) patients. Objective To assess the influence of orexigenic neuropeptides in the nutritional aspects of NAFLD obese adolescents submitted to a long-term interdisciplinary approach. Methods Fifty adolescents aged 15-19 years, with body mass index at least 95th percentile, consisting of 25 patients without NAFLD and 25 with NAFLD. The NAFLD diagnosis was determined by ultrasonography. Blood samples were collected to analyze glycemia, hepatic transaminases, and lipid profile. Insulin resistance was estimated by Homeostasis Model Assessment Insulin Resistance Index. Neuropeptide Y (NPY) and agouti related protein concentrations were measured by enzyme-linked immunosorbent assay. Analyses of food intake were made by 3 days recordatory inquiry. Results At baseline conditions, the patients with NAFLD had significantly higher values of body mass, body mass index, visceral fat, triglycerides, VLDL-C, and hepatic transaminases. After the long-term intervention, they presented a significant reduction in these parameters. In both the groups, it was observed a significant decrease in energy intake, macronutrients and dietetic cholesterol. Only the patients with NAFLD presented a positive correlation between the saturated fatty acids intake and the orexigenic neuropeptides NPY and agouti related protein, and carbohydrate with NPY. Indeed, it was observed a positive correlation between energy intake, lipid (%) and saturated fatty acids with visceral fat accumulation. Conclusion Our findings showed an important influence of diet composition in the orexigenic system, being essential consider that the excessive saturated fatty acids intake could be a determinant factor to increase nonalcoholic fatty liver disease. Eur J Gastroenterol Hepatol 22:557-563 (C) 2010 Wolters Kluwer Health | Lippincott Williams & Wilkins.

Arterial supply of the penis in agoutis (Dasyprocta prymnolpha, Wagler, 1831)

The arterial vascularization of agoutis` penis (Dasyprocta prymnolopha) were analysed using ten male adults from `Nucleo de Estudos e Preservacao de Animais Silvestres da Universidade Federal do Piaui` (FUFPI/IBAMA n degrees 02/99). Among the total number of specimens, six animals had natural death and were members of the research collection of the Laboratory of Anatomy, and four were killed after anaesthesia. Stained bi-centrifugated-Cis-I-4 latex was injected in arterial vessels responsible for penis vascularization throughout the abdominal portion of aorta. The samples were fixed in 10% formaldehyde solution and arteries were dissected. The penile artery is originated as a branch of internal pudendal artery. At the level of ischiatic arch, the penile artery project two branches, the penile dorsal and the deep arteries; those arteries irrigates the penile dorsal surface and the corpus cavernosum penis. The penile dorsal arteries have an independent course up to the glans penis. Based on the conditions of this work a remarkable similarity regarding the distribution of vessels destined to the agouti penis when compared to other domestic, wild and lagomorph rodents as rabbits.

Infecções experimentais na Leishmaniose Tegumentar Americana

Rhésus foram infectados com culturas recentemente isoladas de L. brasiliensis. As inoculações foram feitas por via intradérmica nas arcadas superciliares e, excepcionalmente, na superfícia do nariz. O prazo da incubação foi de 1 a 2 mêses. A marcha da infecção varia conforme as lesões se limitam à pele ou se estendem às mucosas. No primeiro caso, a duração da infecção foi de 3 a 8 meses e terminou sempre pela cura espontânea. Quando as lesões se estendem às mucosas elas persistem e se agravam, enquanto as lesões cutâneas observadas no mesmo animal, regridem, tendendo para cicatrização e cura. Cães também foram injetados com resultados positivos. As inoculações foram praticadas por via intradérmica na superfície do nariz. O prazo de incubação foi de cêrca de 2 mêses e as lesões observadas se limitaram à pele e terminaram pela cura. Uma cotia (Dasyprocta agouti) foi inoculada não se mostrando infectada. Um rhésus, curado de uma infecção pela L. brasiliensis, inoculado de novo com outra amostra do mesmo parasito, não se mostrou infectado. A extensão das lesões às mucosas nas infecções experimentais do rhésus pela L. brasiliensis, fato ainda não veificado na L. trópica, constitue mais um argumento em favor da independência daquela espécie.

Experimental heteroxenous cycle of Lagochilascaris minor Leiper, 1909 (Nematoda: Ascarididae) in white mice and in cats

Reports of natural infections of sylvatic carnivores by adult worms of species similar to Lagochilascaris minor in the Neotropical region led to attempts to estabilish experimental cycles in laboratory mice and in cats. Also, larval development was seen in the skeletal muscle of an agouti (Dasyprocta leporina) infected per os with incubated eggs of the parasite obtained from a human case. In cats, adult worms develop and fertile eggs are expelled in the feces: in mice, larval stages of the parasite develop, and are encapsulate in the skeletal muscle, and in the adipose and subcutaneous connective tissue. From our observations, we conclude that the larva infective for the mouse is the early 3rd stage, while for the final host the infective form is the later 3rd stage. A single moult was seen in the mouse, giving rise to a small population of 4th stage larvae, long after the initial infection.

Wild Rodents as Experimental Intermediate Hosts of Lagochilascaris minor Leiper, 1909

A total of 25 specimens of Cavia porcellus (guinea pig), 5 Dasyprocta agouti (agouti), and 22 Calomys callosus (vesper mice) were inoculated with infective eggs of Lagochilascaris minor. The inoculum was prepared with embryonated eggs and orally administered to each individual animal through an esophagus probe. In parallel, 100 specimens of Felis catus domesticus were individually fed with 55-70 nodules containing 3rd-stage larvae encysted in tissues of infected rodents. Animals were examined and necropsied at different time intervals. The migration and encystment of L3 larva was observed in viscera, skeletal muscle, adipose and subcutaneous tissues from all rodents. Adult worms localized at abscesses in the cervical region, rhino, and oropharynx were recovered from domestic cats inoculated with infected rodent tissues. Through this study we can conclude that: (1) wild rodents act as intermediate hosts, characterizing this ascarid heteroxenic cycle; (2) in natural conditions rodents could possibly act as either intermediate hosts or paratenic hosts of Lagochilascaris minor; (3) despite the occurrence of an auto-infecting cycle, in prime-infection of felines (definite hosts) the cycle is only completed when intermediate hosts are provided; and (4) in the wild, rodents could serve as a source of infection for humans as they are frequently used as food in regions with the highest incidence of human lagochilascariasis.

Pleiotropy in the melanocortin system, coloration and behavioural syndromes.

In vertebrates, melanin-based coloration is often associated with variation in physiological and behavioural traits. We propose that this association stems from pleiotropic effects of the genes regulating the synthesis of brown to black eumelanin. The most important regulators are the melanocortin 1 receptor and its ligands, the melanocortin agonists and the agouti-signalling protein antagonist. On the basis of the physiological and behavioural functions of the melanocortins, we predict five categories of traits correlated with melanin-based coloration. A review of the literature indeed reveals that, as predicted, darker wild vertebrates are more aggressive, sexually active and resistant to stress than lighter individuals. Pleiotropic effects of the melanocortins might thus account for the widespread covariance between melanin-based coloration and other phenotypic traits in vertebrates.

Neuroendocrine characterization and anorexigenic effects of telmisartan in diet- and glitazone-induced weight gain.

Telmisartan is an angiotensin II receptor blocker with peroxisome proliferator-activated receptor-gamma agonistic properties. Telmisartan prevents weight gain and decreases food intake in models of obesity and in glitazone-treated rodents. This study further investigates the influence of telmisartan and pioglitazone and their association on weight gain and body composition by examining their influence on neuroendocrine mediators involved in food intake. Male C57/Black 6 mice were fed a high-fat diet, weight matched, and randomized in 4 treatment groups: vehicle, pioglitazone, telmisartan, and pioglitazone-telmisartan. Weight gain, food and water intake, body composition, plasma leptin levels, and the hypothalamic expression of neuroendocrine mediators were analyzed. Additional studies were performed with irbesartan and in angiotensin II 1(A) receptor-knockout mice. Telmisartan abolished weight and fat gain in vehicle- and pioglitazone-treated mice while decreasing food intake, the hypothalamic expression of the agouti-related protein, and plasma leptin levels. Modifications in neuropeptide Y and proopiomelanocortin were not consistent with changes in food intake. The effects on weight gain and expression of the agouti-related protein were intermediate with irbesartan. The effects of telmisartan on weight gain were even more pronounced in angiotensin II 1(A) receptor-knockout mice. This study confirms the anorexigenic effects of telmisartan in mice fed a high-fat diet and suggests for the first time a functional role of telmisartan on hypothalamic orexigenic agouti-related protein regulation. These anorexigenic properties abolish both weight gain and body composition modifications in fat-fed and glitazone-treated mice. The anorexigenic properties are independent from the angiotensin II 1(A) receptor.

Rôle et mécanisme d'action de la metformine et du telmisartan sur la régulation de la prise alimentaire

Dans ce travail de thèse, nous avons étudié les mécanismes d'action de deux médicaments connus pour diminuer la prise alimentaire et pondérale : la metformine et le telmisartan. Nous avons dans un premier temps étudié les effets de la metformine, un antidiabétique oral connu pour avoir des effets anorexigènes. Les mécanismes hypothalamiques potentiellement impliqués dans la modulation de la prise alimentaire par la metformine ont été étudiés dans trois groupes de rats : un groupe de rats obèses (DIO), un groupe de rats résistants à l'obésité (DR) ainsi qu'un groupe contrôle. A la fin de la période de prise pondérale de six mois, les rats DIO avaient des taux d'ARNm de NPY hypothalamique plus élevés que leurs congénères résistants et contrôles. Chez les DIO ainsi que chez les DR un traitement par metformine induit une baisse significative de la prise alimentaire accompagnée par une baisse du poids. Nous avons pu d'autre part constater que la perte de poids obtenue par un traitement de metformine était corrélée aux taux circulants de leptine avant le traitement. Cet effet s'accompagne d'une augmentation de l'expression du récepteur ObRb au niveau hypothalamique. Dans un second temps, nous avons étudié les effets du telmisartan, un inhibiteur du récepteur à l'angiotensine II ayant une activité agoniste partielle PPARγ. L'influence du telmisartan associé à la pioglitazone sur la prise alimentaire et pondérale a été examinée en étudiant leur effet sur les neuropeptides hypothalamiques responsables du contrôle de la prise alimentaire. Quatre groupes de souris soumises à un régime riche en graisse ont été formés : un groupe placebo, un groupe pioglitazone, un groupe telmisartan et un groupe pioglitazone-telmisartan. Le telmisartan a aboli la prise pondérale induite par une diète riche en graisse ou par un traitement de pioglitazone. Cette diminution était corrélée à une baisse de la prise alimentaire et de l'expression hypothalamique d'AgRP. Cette étude confirme donc les effets anorexigènes du telmisartan et démontre pour la première fois le rôle fonctionnel du telmisartan sur l'expression hypothalamique d'AgRP. English Abstract : In this work, we investigated the effect of two drugs known to have interessants effects on food intake and body weight. First we investigated the hypothalamic mechanisms potentially implicated in the modulation of feeding by the glucose-lowering drug metformin in three different groups of animals: diet-induced obese (DIO) and diet-resistant (DR) male rats as well as lean controls (CT). At the end of the high fat diet period, despite higher leptin levels, DIO rats had higher levels of hypothalamic NPY expression than DR or CT, suggesting a central leptin resistance. In DIO but also in DR rats, metformin treatment induced significant reductions of food intake accompanied by decreases in body weight. Interestingly, the weight loss achieved by metformin was correlated with pre-treatment plasma leptin levels. This effect was paralleled by a stimulation of the expression of the leptin receptor gene (ObRb) in the arcuate nucleus. Next we investigated the antihypertensive drug Telmisartan, an angiotensin II receptor blocker with PPARγ agonistic properties. The influence of telmisartan, of pioglitazone and of their association on weight gain and food intake was assessed by studying their effects on neuro-endocrine mediators involved in food intake. Mice were fed a high fat diet, weightmatched and randomized in four treatment groups: vehicle, pioglitazone, telmisartan and pioglitazone-telmisartan. Telmisartan treatment was found to abolish weight and fat gain in either vehicle or pioglitazone treated mice. This effect was accompanied by a decrease in food intake. The hypothalamic expression of the agouti-related protein and plasma leptin levels show also a decrease under metformin treatment. This study confirms the anorexigenic effects of telmisartan in mice fed a high fat diet, and suggests for the first time a functional role of telmisartan on hypothalamic orexigenic agouti-related protein regulation.

Observations on Eimeria species of Dasyprocta leporina (Linnaeus, 1758)(Rodentia: Dasyproctidae) from the state of Pará, North Brazil

Redescriptions are given of the mature oocysts of Eimeria aguti Carini 1935, E. cotiae Carini, 1935 and E. paraensis Carini, 1935, in the faeces of five specimens of the rodent Dasyprocta leporina (Rodentia: Dasyproctidae) from the state of Pará, North Brazil. New information is provided on the sporulation time of these parasites and the prepatent period in experimentally infected D. leporina. Some endogenous stages of E. cotiae are described in the epithelial cells of the ileum, and the absence of any oocysts in the gall-bladder contents of the infected animals indicates that the intestine is also the site of development of E. aguti and E. paraensis. Difficulties in separating E. cotiae and E. paraensis on morphology of the oocysts are discussed. The oocysts of both parasites share many structural features and have a wide size range. It is concluded that although it is at present best to maintain these names, the possibility exists that they were separately given to oocysts of smaller dimensions (E. cotiae) and larger dimensions (E. paraensis) of a single parasite. Location of an endogenous site of development for E. paraensis that is distinctly separate from that of E. cotiae might establish more definitely the separate specific status of the two parasites.

Eumelanin- and pheomelanin-based colour advertise resistance to oxidative stress in opposite ways.

The control mechanisms and information content of melanin-based colourations are still debated among evolutionary biologists. Recent hypotheses contend that molecules involved in melanogenesis alter other physiological processes, thereby generating covariation between melanin-based colouration and other phenotypic attributes. Interestingly, several molecules such as agouti and glutathione that trigger the production of reddish-brown pheomelanin have an inhibitory effect on the production of black/grey eumelanin, whereas other hormones, such as melanocortins, have the opposite effect. We therefore propose the hypothesis that phenotypic traits positively correlated with the degree of eumelanin-based colouration may be negatively correlated with the degree of pheomelanin-based colouration, or vice versa. Given the role played by the melanocortin system and glutathione on melanogenesis and resistance to oxidative stress, we examined the prediction that resistance to oxidative stress is positively correlated with the degree of black colouration but negatively with the degree of reddish colouration. Using the barn owl (Tyto alba) as a model organism, we swapped eggs between randomly chosen nests to allocate genotypes randomly among environments and then we measured resistance to oxidative stress using the KRL assay in nestlings raised by foster parents. As predicted, the degree of black and reddish pigmentations was positively and negatively correlated, respectively, with resistance to oxidative stress. Our results reveal that eumelanin- and pheomelanin-based colourations can be redundant signals of resistance to oxidative stress.

Evidence for hypothalamic ketone body sensing: impact on food intake and peripheral metabolic responses in mice.

Monocarboxylates have been implicated in the control of energy homeostasis. Among them, the putative role of ketone bodies produced notably during high-fat diet (HFD) has not been thoroughly explored. In this study, we aimed to determine the impact of a specific rise in cerebral ketone bodies on food intake and energy homeostasis regulation. A carotid infusion of ketone bodies was performed on mice to stimulate sensitive brain areas for 6 or 12 h. At each time point, food intake and different markers of energy homeostasis were analyzed to reveal the consequences of cerebral increase in ketone body level detection. First, an increase in food intake appeared over a 12-h period of brain ketone body perfusion. This stimulated food intake was associated with an increased expression of the hypothalamic neuropeptides NPY and AgRP as well as phosphorylated AMPK and is due to ketone bodies sensed by the brain, as blood ketone body levels did not change at that time. In parallel, gluconeogenesis and insulin sensitivity were transiently altered. Indeed, a dysregulation of glucose production and insulin secretion was observed after 6 h of ketone body perfusion, which reversed to normal at 12 h of perfusion. Altogether, these results suggest that an increase in brain ketone body concentration leads to hyperphagia and a transient perturbation of peripheral metabolic homeostasis.

Segmentos anátomo-cirúrgicos arteriais do rim de cutia (Dasyprocta prymnolopha)

Foram estudados 20 pares de rins decutias (Dasyprocta prymnolopha Wagler, 1831), com o objetivo de descrever os segmentos anátomo-cirúrgicos arteriais. As artérias renais foram injetadas com solução de Vinilite corada, e os rins foram submetidos à corrosão ácida para a obtenção dos moldes vasculares. Observou-se que as artérias renais da cutia, sempre únicas, dividiram-se em artéria setorial ventral e artéria setorial dorsal, caracterizando dois setores renais separados por plano avascular. As artérias setoriais penetraram no hilo renal (100% dos casos). Estes vasos deram origem aos ramos segmentares responsáveis pela irrigação de territórios independentes em cada setor, os segmentos arteriais renais. No rim direito foram observados 3 (60%), 4 (35%) e 5 segmentos (5%) no setor arterial ventral e 3 (30%), 4 (45%), 5 (20%) e 6 (5%) segmentos no setor dorsal e, à esquerda, 2(10%), 3 (55%) e 4 (35%) segmentos no setor ventral e 3 (25%), 4 (50%) e 5 (25%) no dorsal. Com base na distribuição arterial nos rins de cutia, observaram-se setores e segmentos arteriais independentes, sendo possível, desta forma, a realização de setoriectomia e segmentectomia nesta espécie.

Estradiol-induced hypophagia is associated with the differential mRNA expression of hypothalamic neuropeptides

Estradiol participates in the control of energy homeostasis, as demonstrated by an increase in food intake and in body weight gain after ovariectomy in rats. In the present study, female Wistar rats (200-230 g, N = 5-15 per group), with free access to chow, were individually housed in metabolic cages. We investigated food intake, body weight, plasma leptin levels, measured by specific radioimmunoassay, and the hypothalamic mRNA expression of orexigenic and anorexigenic neuropeptides, determined by real-time PCR, in ovariectomized rats with (OVX+E) and without (OVX) estradiol cypionate treatment (10 µg/kg body weight, sc, for 8 days). Hormonal and mRNA expression were determined at pre-feeding and 4 h after food intake. OVX+E rats showed lower food intake, less body weight gain and lower plasma leptin levels. In the OVX+E group, we also observed a reduction of neuropeptide Y (NPY), agouti-related protein (AgRP) and cocaine- and amphetamine-regulated transcript (CART) mRNA expression in the arcuate nucleus and a decrease in orexin A in the lateral hypothalamic area (LHA). There was an increase in leptin receptor (LepRb), melanocortin-4 receptor (MC4-R), CART, and mainly corticotropin-releasing hormone (CRH) mRNA in the paraventricular nucleus and LepRb and CART mRNA in the LHA. These data show that hypophagia induced by estradiol treatment is associated with reduced hypothalamic expression of orexigenic peptides such as NPY, AgRP and orexin A, and increased expression of the anorexigenic mediators MC4-R, LepRb and CRH. In conclusion, estradiol decreases food intake, and this effect seems to be mediated by peripheral factors such as leptin and the differential mRNA expression of neuropeptides in the hypothalamus.

Caractérisation et régulation du métabolisme des acides gras dans l’hypothalamus

Un déséquilibre de la balance énergétique constitue la principale cause du développement des pathologies métaboliques telles que l’obésité et le diabète de type 2. Au sein du cerveau, l’hypothalamus joue un rôle primordial dans le contrôle de la prise alimentaire et du métabolisme périphérique via le système nerveux autonome. Ce contrôle, repose sur l’existence de différentes populations neuronales au sein de l’hypothalamus médio-basal (MBH), neurones à neuropeptide Y (NPY)/Agouti-related peptide (AgRP), et neurones a proopiomelanocortine (POMC), dont l’activité est directement modulée par les variations des taux circulants des nutriments tels que le glucose et les acides gras (FA). Alors que les mécanismes de détection et le métabolisme intracellulaire du glucose ont été largement étudiés, l’implication du métabolisme intracellulaire des FA dans leurs effets centraux, est très peu comprise. De plus, on ignore si le glucose, module le métabolisme intracellulaire des acides gras à longue chaine (LCFA) dans le MBH. Le but de notre première étude est, de déterminer l'impact du glucose sur le métabolisme des LCFA, le rôle de l’AMP-activated protein kinase (AMPK), kinase détectrice du statut énergétique cellulaire, et d'établir s’il y a des changements dans le métabolisme des LCFA en fonction de leur structure, du type cellulaire et de la région cérébrale. Nos résultats montrent que le glucose inhibe l'oxydation du palmitate via l’AMPK dans les neurones et les astrocytes primaires hypothalamiques, in vitro, ainsi que dans les explants du MBH, ex vivo, mais pas dans les astrocytes et les explants corticaux. De plus, le glucose augmente l'estérification du palmitate et non de l’oléate dans les neurones et les explants du MBH, mais pas dans les astrocytes hypothalamiques. Ces résultats décrivent le devenir métabolique de différents LCFA dans le MBH, ainsi que, la régulation AMPK - dépendante de leur métabolisme par le glucose dans les astrocytes et les neurones, et démontrent pour la première fois que le métabolisme du glucose et des LCFA est couplé spécifiquement dans les noyaux du MBH, dont le rôle est critique pour le contrôle de l'équilibre énergétique. Le deuxième volet de cette thèse s’est intéressé à déterminer les mécanismes intracellulaires impliqués dans le rôle de la protéine de liaison ACBP dans le métabolisme central des FA. Nous avons démontré que le métabolisme de l’oléate et non celui du palmitate est dépendant de la protéine ACBP, dans les astrocytes hypothalamiques ainsi que dans les explants du MBH. Ainsi, nos résultats démontrent qu’ACBP, protéine identifiée originellement au niveau central, comme un modulateur allostérique des récepteurs GABA, agit comme un régulateur du métabolisme intracellulaire des FA. Ces résultats ouvrent de nouvelles pistes de recherche liées à la régulation du métabolisme des acides gras au niveau central, ainsi que, la nouvelle fonction de la protéine ACBP dans la régulation du métabolisme des FA au niveau du système nerveux central. Ceci aiderait à identifier des cibles moléculaires pouvant contribuer au développement de nouvelles approches thérapeutiques de pathologies telles que l’obésité et le diabète de type 2.

Effect of feed restriction and supplemental dietary fat on gut peptide and hypothalamic neuropeptide mRNA concentrations in growing wethers

The objectives of the present study were 1) to evaluate the effects of supplemental fat and ME intake on plasma concentrations of glucagon-like peptide-1 (GLP-1), cholecystokinin (CCK), glucose-dependent insulinotropic polypeptide, ghrelin, and oxyntomodulin; and 2) to determine the association of these peptides with DMI and the hypothalamic concentration of mRNA for the following neuropeptides: neuropeptide Y (NPY), agouti-related peptide (AgRP), and proopiomelanocortin (POMC). In a completely randomized block design with a 2 x 2 factorial arrangement of treatments, 32 pens with 2 wethers each were restricted-fed (2.45 Mcal/lamb per day) or offered diets ad libitum (n = 16) with or without 6% supplemental fat (n = 16) for a period of 30 d. Dry matter intake was measured daily. On d 8, 15, 22, and 29, BW was measured before feeding, and 6 h after feeding, blood samples were collected for plasma measurement of insulin, GLP-1, CCK, ghrelin, glucose-dependent insulinotropic polypeptide, oxyntomodulin, glucose, and NEFA concentrations. On d 29, blood was collected 30 min before feeding for the same hormone and metabolite analyses. At the end of the experiment, wethers were slaughtered and the hypothalami were collected to measure concentrations of NPY, AgRP, and POMC mRNA. Offering feed ad libitum (resulting in greater ME intake) increased plasma insulin and NEFA concentrations (P = 0.02 and 0.02, respectively) and decreased hypothalamic mRNA expression of NPY and AgRP (P = 0.07 and 0.02, respectively) compared with the restricted-fed wethers. There was a trend for the addition of dietary fat to decrease DMI (P = 0.12). Addition of dietary fat decreased insulin and glucose concentrations (P < 0.05 and 0.01, respectively) and tended to increase hypothalamic mRNA concentrations for NPY and AgRP (P = 0.07 and 0.11, respectively). Plasma GLP-1 and CCK concentrations increased in wethers offered feed ad libitum compared with restricted-fed wethers, but the response was greater when wethers were offered feed ad libitum and had supplemental fat in the diet (fat x intake interaction, P = 0.04). The prefeeding plasma ghrelin concentration was greater in restricted-fed wethers compared with those offered feed ad libitum, but the concentrations were similar 6 h after feeding (intake x time interaction, P < 0.01). Supplemental dietary fat did not affect (P = 0.22) plasma ghrelin concentration. We conclude that insulin, ghrelin, CCK, and GLP-1 may regulate DMI in sheep by regulating the hypothalamic gene expression of NPY, AgRP, and POMC.