Sami Dridi

Mode of leptin action in chicken hypothalamus

While there have been many studies in various species examining the mode of central leptin action on food intake, there is however a paucity of data in birds. We have, therefore, addressed this issue in broiler chickens because this strain was selected for high growth rate, hence high food intake. Continuous infusion of recombinant chicken leptin (8 microg/kg/h) during 6 h at a constant rate of 3 ml/h resulted in a significant reduction (49-57%) of food intake in 3-week-old broiler chickens (P < 0.05). The effect of leptin within the central nervous system (CNS) was mediated via selective hypothalamic neuropeptides. Leptin significantly decreased the expression of its receptor (Ob-R), neuropeptide Y (NPY), orexin (ORX), and orexin receptor (ORXR) (P < 0.05), but not that of agouti-related protein (AgRP) (anabolic/orexigenic effectors) in chicken hypothalamus. However, the catabolic/anorexigenic neuropeptides namely proopiomelanocortin (POMC) and corticotropin-releasing hormone (CRH) mRNA levels remained unchanged after leptin treatment. Despite the absence of leptin effect on AgRP (the antagonist of melanocortin receptor MCR) and POMC (the precursor of alpha-melanocyte stimulating hormone which is a potent agonist for MCR), leptin significantly decreased the expression of MCR-4/5 gene in chicken hypothalamus (P < 0.05) suggesting that leptin acts directly (as ligand) or indirectly (via other ligands) on MCRs to regulate food intake in birds. Additionally, leptin down-regulated the expression of fatty acid synthase (FAS) gene in chicken hypothalamus, indicating an additional pathway of leptin action on food intake such as described for FAS inhibitors. These findings provide new insight into the mechanism of leptin control of food intake in chickens.

Ghrelin modulates fatty acid synthase and related transcription factor mRNA levels in a tissue-specific manner in neonatal broiler chicks.

The endogenous ligand for the growth hormone (GH) secretagogue receptor ghrelin is a peptide secreted by the stomach of mammals and stimulates food intake and enhances adiposity. In avian species, ghrelin is mainly produced by the proventriculus but reduces food intake whereas its effect on lipogenesis in different tissues is unknown. We therefore investigated the effects of a single intravenous injection of 2.8 microg (1 nmol per chick) recombinant chicken ghrelin in neonatal broiler chicks. Besides food intake and plasma corticosterone levels, mRNA levels of the key lipogenic enzyme fatty acid synthase (FAS) and its related transcription factors sterol regulatory element binding protein-1 (SREBP-1) and peroxisome proliferator-activated receptor-gamma (PPARgamma) were determined in diencephalon, liver and quadriceps femoris muscle before, and 15, 30, and 60 min after injection. Chicken ghrelin administration induced a significant short-term (<30 min) reduction in food intake and markedly elevated plasma corticosterone levels. In diencephalon, FAS, SREBP-1 and PPARgamma mRNA levels were significantly increased within 15 min after ghrelin injection. These observations suggest that central fatty acid metabolism is involved in the anorectic effects of ghrelin. In contrast, hepatic mRNA levels of FAS and both transcription factors were significantly reduced within 30 min after ghrelin injection. In muscle, FAS and transcription factor gene expression was very low and not affected by ghrelin. Overall, our results indicate that ghrelin has opposite effects on FAS and transcription factor mRNA amounts with increased levels in diencephalon (central anorectic effect) and decreased levels in liver (peripheral anti-lipogenic effect) in chickens.

Visfatin gene expression in chickens is sex and tissue dependent

The present study investigated the expression of visfatin mRNA in various tissues of male and female broiler chickens. We also studied the effect of leptin, cerulenin, and food deprivation, known effectors of energy balance and insulin action, on visfatin gene expression in chickens. Using reverse transcription polymerase chain reaction (RT-PCR) and Northern blot analysis, we detected chicken visfatin mRNA transcript in the kidney, hypothalamus, gizzard, liver, pancreas, proventriculus, breast and leg muscle, ovary, testis, lung, intestine, adipose tissue, and heart. Expression of the visfatin gene in various tissues of male and female chickens was determined by real-time quantitative PCR and found to be tissue and sex dependent. In both sexes, compared to other tissues, the visfatin gene is highly expressed in the muscle. Females exhibited greater (P<0.001) abundance of visfatin mRNA in adipose tissue compared to males, whereas compared to females, males showed greater (P<0.05) visfatin mRNA abundance in the kidney. Also, the regulation of visfatin gene expression by leptin, cerulenin, and food deprivation is tissue specific. Leptin decreased (P<0.05) visfatin mRNA abundance in the liver and hypothalamus, but not in muscle. In contrast, cerulenin increased (P<0.01) visfatin gene expression in the liver and in muscle, but not in the hypothalamus. Interestingly, visfatin mRNA levels increased (P<0.05) in the liver after 24-h food deprivation, but not in muscle or in the hypothalamus of genetically selected fat and lean line chickens. Our results showed that the visfatin gene is ubiquitously expressed in chickens with greater abundance in muscle, and that it is regulated in a tissue-specific manner by energy balance-related factors.

Relationship between plasma leptin-like protein levels, begging and provisioning in nestling thin-billed prions Pachyptila belcheri

While there have been many studies in various species examining the physiological role of leptin, there are so far no data in free-living seabirds. In the present study, we assess whether leptin is expressed in thin-billed prions (Pachyptila belcheri) and we investigate its relationship with feeding-related parameters including body condition, begging intensities and provisioning rates. We showed by Western Blot analysis using leptin-specific antibody that leptin-like protein (14-16kDa) is expressed in adipose tissue and liver of nestling thin-billed prions. Plasma leptin-like protein levels, determined by RIA, were in the same range (1-3ng/ml) as in other avian species and increased with age. In two breeding seasons, the plasma leptin-like protein levels were negatively correlated with provisioning rates (R=-0.67 and -0.35 in 2003 and 2004, respectively, P<0.05) indicating that endogenous leptin may be an anorexigenic hormone in wild birds. Plasma leptin-like protein levels were positively correlated with begging intensities (R=0.43 and 0.37 in 2003 and 2004, respectively, P<0.05), and this may be because hungry nestling seabird chicks with low body conditions increased their begging intensities. Plasma leptin-like protein levels did not correlate either with plasma triglyceride or glucose levels in thin-billed prions. Overall, these findings show the presence of leptin-like protein in free-living seabirds and provide new insights into its function and its possible role in feeding-associated behaviours.

Feasibility study on using visible-near infrared spectroscopy coupled with factorial discriminant analysis technique to identify sheep milk from different genotypes and feeding systems

Quick detection of differences in raw milk samples due to feeding and genotype is needed for quality control of milk and milk products. This study was undertaken to evaluate the potential of visible (vis) and near infrared (NIR) spectroscopy to discriminate between differences in sheep milk that are attributed to different feedings and genotypes. A mobile, fibre-type, vis-NIR spectrophotometer (Zeiss Corona 45 visnir fibre, Jena, Germany) with a light reflectance measurement range of 306.5–1710.9 nm was used to measure the light reflectance from 50 raw milk samples obtained from two experimental trials in Tunisia. Out of these 50 samples, 30 samples were from the first trial and were obtained equally from two different genotypes and 20 samples were from the second trial and were obtained equally from two different feedings of the same genotype. Milk samples were classified into four different classes, namely, Comisana genotype with pasture feeding (CO), Sicilo–Sarde genotype with pasture feeding (SS), Sicilo–Sarde genotype feeding on Scotch horse bean meal (SSB) and Sicilo–Sarde genotype feeding on soybean meal (SSS). Another milk sample classification based on three classes for three different feedings of one genotype (Sicilo–Sarde) was considered by removing CO. The factorial discriminant analysis (FDA) was applied on the first five principal components obtained from the principal component analysis performed on the vis-NIR spectra in order to classify milk samples into four and three assigned groups. Based on a four-group classification, correct classification (CC) of 92.5% and 95% was observed for the calibration and validation data sets, respectively. For a three-group classification, CC was 100% and 88.9% for the calibration and validation data sets, respectively. Even though the number of samples in the second trial (different feedings) was smaller (20 samples) compared with those in the first trial (30 samples), the FDA provided better discrimination of differences attributed to feeding compared to those attributed to genotypes. It was concluded that vis-NIR spectroscopy coupled with the FDA classification technique is a powerful tool to discriminate between genotypes and feeding differences that exist in raw sheep milk samples.