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.

Confined housing system increased abdominal and subcutaneous fat deposition and gene expressions of carbohydrate response element-binding protein and sterol regulatory element-binding protein 1 in chicken.

Free-range production system is increasingly being used in poultry breeding and feed production in many countries. The objective of the current experiment was to evaluate the effects of different raising systems on fat-related traits and mRNA levels of liver lipogenesis genes in Erlang Mountainous chicken. Each of 10 birds (91 day old) from caged, indoor-floor housed, and free-range housing systems was slaughtered, and fat-related traits, live body weight (BW), subcutaneous fat thickness (SFT), abdominal fat weight (AFW), abdominal fat percentage (AFP), and intramuscular fat content were determined. The mRNA levels of liver X receptor α, carbohydrate response element-binding protein (ChREBP), sterol regulatory element-binding protein-1 (SREBP1), and fatty acid synthase were detected. The caged chicken exhibited significantly higher BW, SFT, and AFW than those of free-ranged chicken (P < 0.05). All the 4 genes had a similar expression pattern, and they showed the highest level in caged chicken, while the lowest level was found in free-ranged chicken. Association analysis indicated that there were significant (P < 0.05) or highly significant (P < 0.01) positive correlations between the mRNA levels of ChREBP, SREBP1, and fat traits of SFT, AFW, and AFP. Thus, we deduced that increased fat deposition in caged chicken was probably induced by increased gene expression of ChREBP and SREBP1 in the liver.

Chickens from lines selected for high and low body weight show differences in fatty acid oxidation efficiency and metabolic flexibility in skeletal muscle and white adipose tissue.

OBJECTIVE: The Virginia lines of chickens have resulted from more than 55 generations of artificial selection for low (LWS) or high (HWS) juvenile body weight. We hypothesized that the relative hyperphagia and greater body weight in juvenile HWS chickens are associated with altered fatty acid oxidation efficiency and metabolic flexibility in tissues associated with energy sensing and storage, and relative cellular hypertrophy in white adipose tissue. METHODS: Hypothalamus, liver, pectoralis major, gastrocnemius, abdominal fat, clavicular fat and subcutaneous fat were collected from the juvenile (56-65 days old) LWS and HWS chickens for metabolic, gene expression and histological assays. RESULTS: The HWS chickens had reduced fatty acid oxidation efficiency in abdominal fat (P<0.0001) and reduced rates of oxidation in abdominal fat and gastrocnemius (P<0.0001) as compared with the LWS. There was reduced citrate synthase activity in white adipose tissue (P<0.0001) and greater metabolic inflexibility in skeletal muscle (P=0.006) of the HWS compared with the LWS. Greater pyruvate dehydrogenase kinase 4 (PDK4) and forkhead box O1A (FoxO1) mRNA were found in skeletal muscle and white adipose tissue of 56-day-old HWS than LWS. Expression of peroxisome proliferator-activated receptor γ (PPARγ) in all adipose tissue depots was greater (P<0.05) in LWS than in HWS chickens. The HWS chickens had larger (P<0.0001) and fewer (P<0.0001) adipocytes per unit area than the LWS. CONCLUSION: Compared with the LWS, the HWS chickens have impaired metabolic flexibility and fatty acid oxidation efficiency due to greater pyruvate dehydrogenase activity to accommodate the influx of acetyl-CoA from fatty acid oxidation in skeletal muscle and adipose tissue. These metabolic adaptations can be linked to differences in gene expression regulation, adipocyte cellularity and body composition between the lines, which may provide valuable insight into metabolic disorders in other species.

Chickens from lines artificially selected for juvenile low and high body weight differ in glucose homeostasis and pancreas physiology.

Artificial selection of White Plymouth Rock chickens for juvenile (day 56) body weight resulted in two divergent genetic lines: hypophagic low weight (LWS) chickens and hyperphagic obese high weight (HWS) chickens, with the latter more than 10-fold heavier than the former at selection age. A study was designed to investigate glucose regulation and pancreas physiology at selection age in LWS chickens and HWS chickens. Oral glucose tolerance and insulin sensitivity tests revealed differences in threshold sensitivity to insulin and glucose clearance rate between the lines. Results from real-time PCR showed greater pancreatic mRNA expression of four glucose regulatory genes (preproinsulin, PPI; preproglucagon, PPG; glucose transporter 2, GLUT2; and pancreatic duodenal homeobox 1, Pdx1) in LWS chickens, than HWS chickens. Histological analysis of the pancreas revealed that HWS chickens have larger pancreatic islets, less pancreatic islet mass, and more pancreatic inflammation than LWS chickens, all of which presumably contribute to impaired glucose metabolism.

Expression profiles of muscle genes in postnatal skeletal muscle in lines of chickens divergently selected for high and low body weight.

Long-term genetic selection for BW has generated high weight select (HWS) and low weight select (LWS) lines of chickens. These lines show an approximate 10-fold difference in BW at selection age (d 56). The objective of this study was to profile the expression of master regulators of early lineage specification (Pax3, Pax7) and myogenic regulatory factors (Myf5, MyoD1, MyoG, and Mrf4) on day of hatch and d 7, 28, and 56 in pectoralis major and gastrocnemius muscles. There was a line × age interaction for expression of all 6 genes in both muscles. In pectoralis major muscle, Pax3, MyoD1, and Mrf4 showed greater expression in LWS than HWS at day of hatch, whereas all 6 genes showed greater expression in HWS than LWS at d 28. In gastrocnemius muscle, Pax3, Myf5, MyoD1, and MyoG showed greater expression in LWS than HWS at day of hatch, whereas Pax7, Myf5, MyoD1, and Mrf4 showed greater expression in HWS than LWS at d 28. At day of hatch there was no difference in fiber number in gastrocnemius muscle between HWS and LWS; however, HWS had greater fiber diameter than LWS. These results indicate that in LWS there is enhanced expression of genes that are necessary for proliferation of progenitor muscle cells and muscle cell differentiation at day of hatch compared with HWS, but by d 28 these genes are expressed greater in HWS than LWS. Thus, long-term selection for growth has altered the pattern of muscle gene expression.

Expression Profiles of Cellular Retinol-binding Protein, Type II (CRBP II) in Erlang Mountainous Chickens.

Cellular retinol-binding protein II (CRBP II) belongs to the family of cellular retinol-binding proteins and plays a major role in absorption, transport, and metabolism of vitamin A. In addition, because vitamin A is correlated with reproductive performance, we measured CRBP II mRNA abundance in erlang mountainous chickens by real-time PCR using the relative quantification method. The expression of CRBP II showed a tissue-specific pattern and egg production rate-dependent changes. The expression was very high (p<0.05) in jejunum and liver, intermediate in kidney, ovary, and oviduct, and lowest (p<0.05) in heart, hypothalamus, and pituitary. In the hypothalamus, oviduct, ovary, and pituitary, CRBP II mRNA abundance were correlated to egg production rate, which increased from 12 wk to 32 wk, peaked at 32 wk relative to the other time points, and then decreased from 32 wk to 45 wk. In contrast, the expression of CRBP II mRNA in heart, jejunum, kidney, and liver was not different at any of the ages evaluated in this study. These data may help to understand the genetic basis of vitamin A metabolism, and suggest that CRBP II may be a candidate gene to affect egg production traits in chickens.

Effect of hybridization on carcass traits and meat quality of erlang mountainous chickens.

Native chickens hold a significant share of the market in China. In response to the huge demand from the market, the productivity of Chinese native chickens needs to be improved. Cross breeding is an effective method to increase productivity, although it might affect meat quality. In this study, two pure lines (SD02 and SD03) of Erlang mountainous chickens were hybridized with a yellow feather and faster growing line (SD01). The effect of hybridization on carcass and meat quality (physiochemical and textural traits) was measured in the F1 population at d 91 of age. The hybrids exhibited higher body weight and dressed weight, and amount of semi-eviscerated, eviscerated, breast muscle and abdominal fat (p<0.05). Abdominal fat yield also increased (p<0.05) compared to the offspring of the two pure-lines. Meanwhile, there was no significant difference in meat quality traits except for the myofiber diameter and density and the shear force of the breast muscle. Overall, the offspring of cross-lines were similar to pure lines in meat color, pH value, inosinic acid, crude protein, crude fat, dry matter, moisture content and amino acid composition in the breast muscle. These results suggest that productivity can be improved via cross-breeding while maintaining meat quality of the Erlang mountainous chicken.

Housing system affects broiler characteristics of local Chinese breed reciprocal crosses.

Artificial selection in closed populations can fix or differentiate alternative alleles of loci associated with selected traits. Two closed chicken populations, A and B, originating from Jiuwu, a Chinese native breed, were bred for more than 10 generations. We compared progeny from reciprocal crosses (AB and BA) under free range (trial 1) and cage and pen (trial 2) systems. Traits measured included feed conversion, live BW, subcutaneous fat thickness, percentages of carcass, semi-eviscerated carcass, eviscerated carcass, breast muscle (pectoralis major and minor), leg muscle (boneless drum and thigh), heart, gizzard, proventriculus, liver, comb, and abdominal fat, plus moisture and fat content in breast muscle at 91 d. In trial 1, there were no significant differences between crosses for any trait except percentage proventriculus (AB > BA). Males were significantly heavier with greater proportions of semi-eviscerated carcass, leg muscle, heart, and comb than females, whereas females had thicker subcutaneous fat and higher percentage abdominal fat, breast muscle, gizzard, and proventriculus than males. In trial 2, chickens raised in cages were significantly heavier, had superior feed efficiency, thicker subcutaneous fat, higher fat content and percentage abdominal fat, carcass, semi-eviscerated carcass, liver, and comb than those reared in pens. Those reared in pens exhibited significantly greater proportions of breast muscle, gizzard, and proventriculus than those raised in cages. The only significant 2-way interaction was cross × sex for percentage semi-eviscerated and eviscerated carcass, because BA males were greater than other combinations. Interactions of cross × sex × housing system for percentage liver and abdominal fat were significant; cage-reared AB females displayed higher percentages than pen-reared BA males. Results implied that subpopulations should be considered rather than a single larger population as a way to reduce loss of genetic variation in local and heritage stocks. Although performance of reciprocal crosses was similar across housing systems, caging during the finishing stage of a slow-growing breed enhanced feed efficiency, BW, and meat composition.

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.

An antimicrobial peptide is downregulated in the small intestine of Eimeria maxima-infected chickens.

Avian coccidiosis is a major disease of poultry caused by the intestinal protozoa Eimeria. Infection leads to reduced feed efficiency and BW gain, resulting in severe economic losses for the poultry industry. Aviagen line A and line B birds show a differential response to Eimeria infection, with line B birds exhibiting higher lesion scores and mortality. The objective of this study was to examine differential intestinal gene expression between 2-wk-old line A and B chicks in response to a challenge with Eimeria maxima. After challenge with 1 × 10(4) oocysts/chick, more than 40% of line A chicks had lesion scores of 0 to 1 (scale of 0 to 4), similar to control chicks. In contrast, all line B chicks challenged at this same dose had lesion scores of 2 to 4. Total RNA was extracted from the jejunum of control and challenged chicks from both lines A and B. Microarray analysis revealed that liver-expressed antimicrobial peptide 2 (LEAP-2), a component of the innate immune system, was downregulated 20-fold in line A challenged chicks with lesion scores of 2 to 4 compared with line A control chicks, and was downregulated 11- to 71-fold in line B challenged chicks with lesion scores of 2 to 4 compared with line B control chicks. Liver-expressed antimicrobial peptide 2 was downregulated less than 2-fold in line A challenged chicks with lesion scores of 1 compared with line A control chicks, indicating that these chicks were similar to control chicks in their expression level of LEAP-2. Other genes (cytochrome P450, heat shock protein 25, keratin 19, and amino acid transporter ASCT1) showed different patterns of over- or underexpression. The expression of LEAP-2 was verified using real-time PCR, revealing a correlation between lesion score and magnitude of LEAP-2 downregulation for both line A and line B chicks. Thus, LEAP-2 may serve as a useful marker for identification of chickens resistant to E. maxima infection and potentially other Eimeria spp.

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.

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.

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.

Expression profiling of the solute carrier gene family in chicken intestine from the late embryonic to early post-hatch stages.

Intestinal development during late embryogenesis and early post-hatch has a long-term influence on digestive and absorptive capacity in chickens. The objective of this research was to obtain a global view of intestinal solute carrier (SLC) gene family member expression from late embryogenesis until 2 weeks post-hatch with a focus on SLC genes involved in uptake of sugars and amino acids. Small intestine samples from male chicks were collected on embryonic days 18 (E18) and 20 (E20), day of hatch and days 1, 3, 7 and 14 post-hatch. The expression profiles of 162 SLC genes belonging to 41 SLC families were determined using Affymetrix chicken genome microarrays. The majority of SLC genes showed little or no difference in level of expression during E18-D14. A number of well-known intestinal transporters were upregulated between E18 and D14 including the amino acid transporters rBAT, y(+)LAT-2 and EAAT3, the peptide transporter PepT1 and the sugar transporters SGLT1, GLUT2 and GLUT5. The amino acid transporters CAT-1 and CAT-2 were downregulated. In addition, several glucose and amino acid transporters that are novel to our understanding of nutrient absorption in the chicken intestine were discovered through the arrays (SGLT6, SNAT1, SNAT2 and AST). These results represent a comprehensive characterization of the expression profiles of the SLC family of genes at different stages of development in the chicken intestine and lay the ground work for future nutritional studies.

Developmental regulation of nutrient transporter and enzyme mRNA abundance in the small intestine of broilers.

The objective of this study was to investigate intestinal nutrient transporter and enzyme mRNA in broilers selected on corn- and soybean-based (line A) or wheat-based (line B) diets. We investigated the peptide transporter PepT1, 10 amino acid transporters (rBAT, b(o,+)AT, ATB(o,+), CAT1, CAT2, LAT1, y(+)LAT1, y(+)LAT2, B(o)AT, and EAAT3), 4 sugar transporters (SGLT1, SGLT5, GLUT5, and GLUT2), and a digestive enzyme (aminopeptidase N). Intestine was collected at embryo d 18 and 20, day of hatch, and d 1, 3, 7, and 14 posthatch. The mRNA abundance of each gene was assayed using real-time PCR and the absolute quantification method. For PepT1, line B had greater quantities of mRNA compared with line A (P = 0.001), suggesting a greater capacity for absorption of amino acids as peptides. Levels of PepT1 mRNA were greatest in the duodenum (P < 0.05), whereas the abundances of SGLT1, GLUT5, and GLUT2 mRNA were greatest in the jejunum (P < 0.05). Abundances of EAAT3, b(o,+)AT, rBAT, B(o)AT, LAT1, CAT2, SGLT5, and aminopeptidase N mRNA were greatest in the ileum (P < 0.05). Quantities of PepT1, EAAT3, B(o)AT, SGLT1, GLUT5, and GLUT2 mRNA increased linearly (P < 0.01), whereas CAT1, CAT2, y(+)LAT1, and LAT1 mRNA decreased linearly (P < 0.05) with age. Abundance of y(+)LAT2 mRNA changed cubically (P = 0.002) with peaks of expression at day of hatch and d 7, and aminopeptidase N and SGLT5 mRNA changed quadratically (P = 0.005) with age. These results provide a comprehensive profile of the temporal and spatial expression of nutrient transporter mRNA in the small intestine of chicks.

Differential expression of appetite-regulating genes in avian models of anorexia and obesity.

Chickens from lines that have been selected for low (LWS) or high (HWS) juvenile body weight for more than 57 generations provide a unique model by which to research appetite regulation. The LWS display different severities of anorexia, whereas all HWS become obese. In the present study, we measured mRNA abundance of various factors in appetite-associated nuclei in the hypothalamus. The lateral hypothalamus (LHA), paraventricular nucleus (PVN), ventromedial hypothalamus (VMH), dorsomedial nucleus (DMN) and arcuate nucleus (ARC) were collected from 5 day-old chicks that were fasted for 180 minutes or provided with continuous access to food. Fasting increased neuropeptide Y receptor subtype 1 (NPYR1) mRNA in the LHA and c-Fos in the VMH, at the same time as decreasing c-Fos in the LHA, neuropeptide Y receptor subtype 5 and ghrelin in the PVN, and neuropeptide Y receptor subtype 2 in the ARC. Fasting increased melanocortin receptor subtype 3 (MC3R) expression in the DMN and NPY in the ARC of LWS but not HWS chicks. Expression of NPY was greater in LWS than HWS in the DMN. neuropeptide Y receptor subtype 5 mRNA was greater in LWS than HWS in the LHA, PVN and ARC. Expression of orexin was greater in LWS than HWS in the LHA. There was greater expression of NPYR1, melanocortin receptor subtype 4 and cocaine- and amphetamine-regulated transcript in HWS than LWS and mesotocin in LWS than HWS in the PVN. In the ARC, agouti-related peptide and MC3R were greater in LWS than HWS and, in the VMH, orexin receptor 2 and leptin receptor were greater in LWS than HWS. Greater mesotocin in the PVN, orexin in the LHA and ORXR2 in the VMH of LWS may contribute to their increased sympathetic tone and anorexic phenotype. The results of the present study also suggest that an increased hypothalamic anorexigenic tone in the LWS over-rides orexigenic factors such as NPY and AgRP that were more highly expressed in LWS than HWS in several nuclei.

Stress-induced suppression of neuropeptide Y-induced hunger in anorexic chicks involves corticotrophin-releasing factor signalling and the paraventricular nucleus of the hypothalamus.

The Virginia lines of chickens have been selected for low (LWS) or high (HWS) juvenile body weight and have different severities of anorexia and obesity, respectively. The LWS that are exposed to stressors at hatch are refractory to neuropeptide Y (NPY)-induced food intake and the objective of the present study was to determine the underlying mechanisms. Chicks were exposed to a stressor (-20°C for 6 minutes and 22°C and delayed access to food for 24 hours) after hatching and the hypothalamic nuclei, including the lateral hypothalamus (LH), paraventricular nucleus (PVN), ventromedial hypothalamus (VMH) and arcuate nucleus (ARC), were collected 5 days later. In LWS but not HWS, stress exposure up-regulated corticotrophin-releasing factor (CRF), CRF receptor subtypes 1 and 2 (CRFR1 and CRFR2, respectively), melanocortin receptor 4 and urocortin 3 in the PVN, as well as CRFR2 mRNA in the VMH and ARC. In LWS, stress exposure was also associated with greater NPY and NPY receptor subtype 5 mRNA in the ARC and PVN, respectively, as well as decreased agouti-related peptide mRNA in the ARC. In HWS, stress exposure was associated with increased CRFR1 and decreased cocaine- and amphetamine-regulated transcript in the ARC and PVN, respectively. Refractoriness of the food intake response to NPY in LWS may thus result from the over-riding anorexigenic tone in the PVN associated with CRF signalling. Indeed, the orexigenic effect of NPY was restored when LWS were injected with a CRF receptor antagonist, astressin, before stress exposure. The results of the present study provide insights into the molecular basis of eating disorders and suggest that CRF signalling in the PVN may exacerbate the anorexic phenotype in the presence of environmental stressors.

Anorexia is Associated with Stress-Dependent Orexigenic Responses to Exogenous Neuropeptide Y.

Chicken lines that have been divergently selected for either low (LWS) or high (HWS) body weight at 56 days of age for more than 57 generations have different feeding behaviours in response to a range of i.c.v. injected neurotransmitters. The LWS have different severities of anorexia, whereas the HWS become obese. Previously, we demonstrated that LWS chicks did not respond, whereas HWS chicks increased food intake, after central injection of neuropeptide Y (NPY). The present study aimed to determine the molecular mechanisms underlying the loss of orexigenic function of NPY in LWS. Chicks were divided into four groups: stressed LWS and HWS on day of hatch, and control LWS and HWS. The stressor was a combination of food deprivation and cold exposure. On day 5 post-hatch, each chick received an i.c.v. injection of vehicle or 0.2 nmol of NPY. Only the LWS stressed group did not increase food intake in response to i.c.v. NPY. Hypothalamic mRNA abundance of appetite-associated factors was measured at 1 h post-injection. Interactions of genetic line, stress and NPY treatment were observed for the mRNA abundance of agouti-related peptide (AgRP) and synaptotagmin 1 (SYT1). Intracerebroventricular injection of NPY decreased and increased AgRP and SYT1 mRNA, respectively, in the stressed LWS and increased AgRP mRNA in stressed HWS chicks. Stress was associated with increased NPY, orexin receptor 2, corticotrophin-releasing factor receptor 1, melanocortin receptor 3 (MC3R) and growth hormone secretagogue receptor expression. In conclusion, the loss of responsiveness to exogenous NPY in stressed LWS chicks may be a result of the decreased and increased hypothalamic expression of AgRP and MC3R, respectively. This may induce an intensification of anorexigenic melanocortin signalling pathways in LWS chicks that block the orexigenic effect of exogenous NPY. These results provide insights onto the anorexic condition across species, and especially for forms of inducible anorexia such as human anorexia nervosa.

Complex spatial and temporal variation of subtropical benthic macrofauna under sewage impact.

To better understand variation in sewage-impacted benthic macrofauna from subtropical tidal flats over time and space, we applied a five-factor linear model at a hierarchy of spatial (Condition - Contaminated or Non-Contaminated, Tidal Flat and Plot) and temporal scales (Season and Fortnight). The Contaminated site showed high levels of coprostanol and the presence of Paranais cf frici as markers or indicators of organic enrichment. Differences between Seasons were more pronounced for the faunal variation patterns than for the other parameters, with lower species richness and abundance in summer. There were significant interactions between Fortnight and Tidal Flat for most variables, reflecting marked heterogeneity within Tidal Flats. Benthic community has significantly changed over short periods of time. These rapid changes may lead to erroneous interpretations and mask the true sources of variation. Our results clearly demonstrate the need to better understand benthic temporal variability even at small scales.