Epigenetic modifiers identified as regulators of food intake in a unique hypophagic chicken model.

DNA methylation is an epigenetic modification that influences gene transcription; however, the effects of methylation-influencing chemicals on appetite are unknown. We evaluated the effects of single administration of a methyl donor, S-Adenosylmethionine (SAM), or methylation inhibitor, 5-Azacytidine (AZA), on immediate and later-age food intake in an anorexic chick model. The doses of intracerebroventricularly-injected SAM were 0 (vehicle), 0.1, 1, and 10 µg, and of AZA were 0 (vehicle), 1, 5, and 25 µg. When injected on day 5 posthatch, there was no effect of SAM on food intake in either fed or fasted chicks, whereas AZA increased food consumption in the fasted state but decreased it in fed chicks. We then performed a single injection (same doses) at hatch and measured food intake on day 5 in response to neuropeptide Y (NPY; 0.2 µg) injection. Irrespective of NPY, chicks injected with 1 µg of SAM ate more than others on day 5. In contrast, chicks injected with AZA (5 and 25 µg doses) consumed less on day 5. In conclusion, we identified DNA methylation-regulating chemicals as regulators of food intake. AZA but not SAM affected food intake in the short-term, feeding state dependently. Later, both chemicals injected on the day of hatch were associated with food intake changes at a later age, suggesting that feeding pathways might be altered through changes in methylation.

The anorexigenic effect of vasoactive intestinal polypeptide in Japanese quail is associated with molecular changes in the arcuate and dorsomedial hypothalamic nuclei.

Vasoactive intestinal polypeptide (VIP) is involved in gastric smooth muscle relaxation, vasodilation, and gastric secretions. It is also associated with appetite regulation, eliciting an anorexigenic response in mammals, birds, and fish; however, the molecular mechanism mediating this response is not well understood. The aim of the present study was thus to investigate hypothalamic mechanisms mediating VIP-induced satiety in 7-d old Japanese quail. In experiment 1, chicks that received intracerebroventricular (ICV) injection of VIP had reduced food intake for up to 180 min after injection and reduced water intake for 90 min. In experiment 2, VIP-treated chicks that were food restricted did not reduce water intake. In experiment 3, there was increased c-Fos immunoreactivity in the arcuate (ARC) and dorsomedial (DMN) nuclei of the hypothalamus in VIP-injected quail. In experiment 4, ICV VIP was associated with decreased neuropeptide Y mRNA in the ARC and DMN and an increase in corticotropin releasing factor mRNA in the DMN. In experiment 5, VIP-treated chicks displayed fewer feed pecks and locomotor behaviors. These results demonstrate that central VIP causes anorexigenic effects that are likely associated with reductions in orexigenic tone involving the ARC and DMN.

The anorexigenic effect of adrenomedullin in Japanese quail (Coturnix japonica) involves increased proopiomelanocortin and cocaine- and amphetamine-regulated transcript mRNAs in the arcuate nucleus of the hypothalamus.

Central administration of adrenomedullin (AM), a 52-amino acid peptide, is associated with anorexigenic effects in some species, including rodents and chickens. However, the associated hypothalamic mechanisms remain unclear and it is unknown if this peptide exerts satiety-inducing effects in other avian species. The objective of this study was thus to investigate AM-induced anorexigenic effects in 7-day-old Japanese quail (Coturnix japonica). After intracerebroventricular injection of 0.3, 1.0, or 3.0 nmol of AM, quail injected with 3.0 nmol of AM ate and drank less than vehicle-injected quail at 180 min after injection. Except for the 1.0 nmol dose of AM exerting an anorexigenic effect at 90 min after injection, no other inhibitory effects on food or water intake were observed. At 60 min after injection, the AM-injected quail had more c-Fos immunoreactive cells in the arcuate nucleus (ARC) than vehicle-injected birds. In the ARC, AM injection was associated with increased proopiomelanocortin (POMC) and cocaine- and amphetamine-regulated transcript (CART) mRNAs. In conclusion, the results suggest that the anorexigenic effect of AM is possibly influenced by the synergistic effect of POMC and CART in the ARC.

Prolactin-releasing peptide increases food intake and affects hypothalamic physiology in Japanese quail (Coturnix japonica).

Prolactin-releasing peptide (PrRP) increases food intake in birds, whereas it is a potent satiety factor in rodents and fish. The aim of this study was to determine the effects of central injection of PrRP on feeding behaviors and hypothalamic physiology in juvenile Japanese quail (Coturnix japonica). Intracerebroventricular injection of 1,692 pmol of PrRP increased food intake for the first 90 min after injection but did not affect water intake. Quail treated with PrRP displayed more food and drink pecks, less time standing but more perching, and decreased defecations. Prolactin-releasing peptide-injected quail had increased c-Fos immunoreactivity in the dorsomedial nucleus (DMN) and arcuate nucleus (ARC) of the hypothalamus. Hypothalamic neuropeptide Y receptor subtypes 2 and 5 and melanocortin receptor 4 mRNAs were greater in PrRP- than vehicle-injected quail. In the DMN, there was less corticotropin-releasing factor (CRF) mRNA and in the ARC, more CRF mRNA in PrRP- than vehicle-injected chicks. Thus, PrRP increases food intake in quail, which is associated with changes in hypothalamic CRF and neuropeptide Y receptor gene expression and c-Fos-immunolabeled cells in the ARC and DMN.

Uptake of manganese from manganese-lysine complex in the primary rat intestinal epithelial cells.

This study was conducted to compare the differences of the uptake of Mn from Mn-lysine complex (MnLys) and MnSO4 and to determine the potential mechanisms of the uptake of Mn from MnLys. We first established the primary rat intestinal epithelial cell culture model and used it to determine the uptake of Mn from different Mn sources. The MnLys increased (p < 0.001) Mn uptake when compared to MnSO4 . The uptake of Mn decreased (p < 0.05) with added Fe concentration increasing in the medium regardless of Mn source. The MnLys decreased (p < 0.01) Mn2+ efflux transporter ferroportin 1 (FPN1) mRNA level, but did not influence (p > 0.06) Mn2+ influx transporter DMT1 mRNA expression when compared to MnSO4 . The results above indicated that the increase of Mn accumulation for MnLys at least partly was due to the decrease of Mn efflux by reduced FPN1 expression. The N-ethylmaleimide, as an l-lysine transport system y+ inhibitor, decreased (p < 0.001) the uptake of Mn from MnLys, but did not affect (p > 0.10) the uptake of Mn from MnSO4 . The cycloheximide, as an l-lysine transport system b0,+ activator, increased (p < 0.001) the uptake of Mn from MnLys, whereas also did not influence the uptake of Mn from MnSO4 . The MnLys increased (p < 0.01) the system y+ member cationic amino acid transporter (CAT) 1, and system b0,+ components rBAT and b0,+ AT mRNA expression when compared to MnSO4 . These results suggested that the uptake of Mn from MnLys complex might be transported by CAT1 and system b0,+ , which was different from ionized Mn2+ uptake pathway. In conclusion, the uptake of MnLys complex not only might be absorbed as Mn2+ , but also appeared to be transported through CAT1 and system b0,+ in the primary rat intestinal epithelial cells.

Dietary protein composition influences abundance of peptide and amino acid transporter messenger ribonucleic acid in the small intestine of 2 lines of broiler chicks.

This study evaluated the effect of dietary protein composition on mRNA abundance of a peptide transporter (peptide transporter 1, PepT1), amino acid (AA) transporters [Na(+)-independent cationic and zwitterionic AA transporter (b(o,+)AT), excitatory AA transporter 3 (EAAT3), Na(+)-independent cationic and Na(+)-dependent neutral AA transporter 2 (y(+)LAT2), L-type AA transporter 1 (LAT1), and cationic AA transporter 1 (CAT1)], and a digestive enzyme (aminopeptidase N) in 2 lines (A and B) of broilers that differentially express PepT1 mRNA (line B > line A). From d 8 to 15 posthatch, birds were fed 1 of 3 diets. Protein sources included whey protein concentrate, a whey partial hydrolysate (WPH), or a mixture of free AA (AA) identical to the composition of whey. Quantities of mRNA were assayed by real-time PCR in the small intestine of males at d 8, 9, 11, 13, and 15. For all genes except LAT1, abundance of mRNA was greatest in line B birds that consumed the WPH diet (P < 0.006). When mRNA abundance was normalized to beta-actin quantities, this effect disappeared, demonstrating a generalized effect on gene expression in line B birds that consumed the hydrolysate. There was a greater villus height:crypt depth ratio (P < 0.05) in line B birds fed the WPH diet as compared with line A. In conclusion, line B birds, which express greater PepT1, displayed enhanced intestinal mucosal absorptive surface area and differential regulation of PepT1, AA transporters, and aminopeptidase N in response to dietary protein composition.

Flavonoids influence epigenetic-modifying enzyme activity: structure - function relationships and the therapeutic potential for cancer.

Epigenetic modifications result in heritable changes in gene expression without changes to the DNA sequence. The most common forms of epigenetic regulation of gene expression are DNA methylation and histone acetylation or methylation, all of which are associated with chromatin remodeling. Results from recent studies suggest that epigenetic changes are some of the primary contributory factors of tumor-suppressor gene silencing in cancer cells. Compounds that target epigenetic regulators in the body may represent an attractive target for chemoprevention. Flavonoids are polyphenolic phytochemicals that exert a multitude of beneficial effects on human health. In recent years, isoflavones, flavonols and catechins have received much attention due to their ability to influence activity of chromatin-modifying enzymes. Epigallocatechin-3-gallate, for example, was shown to inhibit activity of histone acetyltransferase and DNA methyltransferase. In this review, we will highlight the structure-function relationship between flavonoids and epigenetic modifications, with an emphasis on the isoflavones, flavonols and catechins, and their potential as anti-cancer agents in this regard.

Board-invited review: Peptide absorption and utilization: Implications for animal nutrition and health.

Over the last 50 yr, the study of intestinal peptide transport has rapidly evolved into a field with exciting nutritional and biomedical applications. In this review, we describe from a historical and current perspective intestinal peptide transport, the importance of peptides to whole-body nutrition, and the cloning and characterization of the intestinal peptide transporter, PepT1. We focus on the nutritional significance of peptide transport and relate these findings to livestock and poultry. Amino acids are transported into the enterocyte as free AA by a variety of AA transporters that vary in substrate specificity or as di- and tripeptides by the peptide transporter, PepT1. Expression of PepT1 is largely restricted to the small intestine in most species; however, in ruminants, peptide transport and activity is observed in the rumen and omasum. The extent to which peptides are absorbed and utilized is still unclear. In ruminants, peptides make a contribution to the portal-drained visceral flux of total AA and are detected in circulating plasma. Peptides can be utilized by the mammary gland for milk protein synthesis and by a variety of other tissues. We discuss the factors known to regulate expression of PepT1 including development, diet, hormones, diurnal rhythm, and disease. Expression of PepT1 is detected during embryological stages in both birds and mammals and increases with age, a strategic event that allows for the immediate uptake of nutrients after hatch or birth. Both increasing levels of protein in the diet and dietary protein deficiencies are found to upregulate the peptide transporter. We also include in this review a discussion of the use of dietary peptides and potential alternate routes of nutrient delivery to the cell. Our goal is to impart to the reader the nutritional implications of peptide transport and dietary peptides and share discoveries that shed light on various biological processes, including rapid establishment of intestinal function in early neonates and maintenance of intestinal function during fasting, starvation, and disease states.