RESUMEN
Chickens selected for low (LWS) and high (HWS) juvenile body weight (BW) for 55 generations differ in BW by 10-fold at selection age. High (HWR) and low (LWR) body weight-relaxed lines have been random-bred since the 46th generation. Our objective was to evaluate the developmental and nutritional regulation of pancreatic mRNA abundance of pancreatic and duodenal homeobox 1 (PDX1), preproinsulin (PPI), preproglucagon (PPG), and glucose transporter 2 (GLUT2). At day of hatch (DOH) and days 1, 3, 7, and 15 (D1, 3, 7 and 15, respectively), pancreas was collected and real time PCR was performed in Experiment 1. In Experiment 2, HWS and LWS were fed or delayed access to food for 72 h post-hatch, and pancreas collected at D15. There was an interaction of line and age for GLUT2 (P=0.001), PPI (P<0.0001), PPG (P=0.034), and PDX1 (P<0.0001). Expression was greater in chicks from LWR and LWS than HWR and HWS. There was an interaction of line and nutrition on PPG (P<0.0001) and GLUT2 (P=0.001) mRNA, where expression was similar among chicks that were fed but greater in LWS than HWS when chicks were delayed access to food. Thus, the first two weeks is important for maturation of pancreatic endocrine function. Long-term selection for BW is associated with differences in pancreas development, and delaying access to food at hatch may have persisting effects on glucose regulatory function.
Asunto(s)
Proteínas Aviares/genética , Peso Corporal/genética , Pollos/genética , Regulación del Desarrollo de la Expresión Génica , Glucagón/genética , Transportador de Glucosa de Tipo 2/genética , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Animales Recién Nacidos , Cruzamiento , Pollos/crecimiento & desarrollo , Femenino , Alimentos , Masculino , Páncreas/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Selección Genética , Factores de Tiempo , Regulación hacia Arriba/genéticaRESUMEN
Long-term selection for juvenile body weight from a common founder population resulted in two divergent chicken lines (low-weight selected line (LWS), high-weight selected line (HWS)) that display distinct food intake and blood glucose responses to exogenous neuropeptides and insulin. The objective of this study was to elucidate putative targets affecting food intake and energy homeostasis by sequencing hypothalamic RNA from LWS and HWS chickens after insulin injection. Ninety-day-old female LWS and HWS chickens were injected with either vehicle or insulin and hypothalamus collected at 1 h postinjection. Through RNA sequencing, a total of 361 differentially expressed genes (DEGs) were identified. There was greater expression of genes, mainly tyrosine hydroxylase (TH), L-aromatic amino acid decarboxylase (DDC), and vesicular monoamine transporter (VMAT), involved in serotonin and dopamine biosynthesis and signaling in LWS than in HWS vehicle-injected chickens. In contrast, after insulin injection, these genes were more highly expressed in HWS than in LWS. We identified 90 single nucleotide polymorphisms (SNPs) existing only in the HWS and 121 SNPs specific to LWS and 5119 SNPs close to fixation (with absolute frequency difference ≥0.9). Four were located in genes encoding enzymes associated with serotonergic and dopaminergic pathways, such as DDC, TH, and solute carrier family 18, member 2 (VMAT). These data implicate differences in biogenic amines such as serotonin and dopamine in hypothalamic physiology between the chicken lines, and these differences might be associated with polymorphisms during long-term selection. Changes in serotonergic and dopaminergic signaling pathways in response to insulin injection suggest a role in whole-body energy homeostasis.
Asunto(s)
Monoaminas Biogénicas/biosíntesis , Expresión Génica , Hipotálamo/metabolismo , Insulina/farmacología , Transducción de Señal/genética , Animales , Peso Corporal/genética , Pollos/genética , Ingestión de Alimentos/genética , Femenino , Homeostasis/genética , Hipotálamo/efectos de los fármacos , Polimorfismo de Nucleótido Simple , Transducción de Señal/efectos de los fármacosRESUMEN
This experiment used 2 lines of chickens that have been selected 54 generations for either low (LWS) or high (HWS) 8-wk BW from the same founder population, sublines (HWR and LWR) in which selection was relaxed in generation 43 in the selected lines, and crosses (HL and LH) made from generation 54 of HWS and LWS. For 8-wk BW, the difference between lines LWS and HWS in generation 54 was approximately 10-fold, whereas for the relaxed contemporary lines they were approximately 7-fold. Three trials were designed to measure developmental, nutritional, and genetic aspects of blood glucose homeostasis during the first 2 wk posthatch. In trial 1, we measured BW, whole blood glucose (BG), and weights (relative to BW) of liver, pancreas, and yolk sac of chicks fed from day of hatch to d 15. In trial 2, we compared those traits in chicks feed-delayed 72 h posthatch and in chicks without feed delay. In trial 3, we evaluated the effect of a 16-h fast on BW and BG on d 3, 8, and 15. There were higher levels of BG in HWS than LWS, and males than females in the fed state. Delayed access to feed for 72 h after hatch was associated with a dramatic reduction in BG. Feeding triggered a compensatory response whereby LWS displayed greater BG but smaller pancreases (% BW; d 15), compared with the controls. There were maternal effects for BW in both fed and fasted states and the reciprocal crosses exhibited heterosis for BG in the fasted state. These results show that chickens selected for high or low BW differ in BG regulation during the early posthatch period.
Asunto(s)
Glucemia/análisis , Pollos/crecimiento & desarrollo , Pollos/genética , Privación de Alimentos/fisiología , Selección Genética , Factores de Edad , Animales , Peso Corporal , Pollos/metabolismo , Femenino , Homeostasis , Hígado/crecimiento & desarrollo , Masculino , Tamaño de los Órganos , Páncreas/crecimiento & desarrollo , Factores Sexuales , Factores de Tiempo , Saco Vitelino/crecimiento & desarrolloRESUMEN
Chickens from lines selected for low (LWS) or high (HWS) body weight differ by 10-fold in body weight at 56 days old with differences in food intake, glucose regulation, and body composition. To evaluate if there are differences in appetite-regulatory factor and glucose transporter (GLUT) mRNA that are accentuated by hypoglycemia, blood glucose was measured, and hypothalamus, liver, pectoralis major, and abdominal fat collected at 90 days of age from female HWS and LWS chickens, and reciprocal crosses, HL and LH, at 60 min after intraperitoneal injection of insulin. Neuropeptide Y (NPY) and receptor (NPYR) subtypes 1 and 5 mRNA were greater in LWS compared with HWS hypothalamus (P < 0.05), but greater in HWS than LWS in fat (P < 0.05). Expression of NPYR2 was greater in LWS than HWS in pectoralis major (P < 0.05). There was greater expression in HWS than LWS for GLUT1 in hypothalamus and liver (P < 0.05), GLUT2 in fat and liver (P < 0.05), and GLUT9 in liver (P < 0.05). Insulin was associated with reduced blood glucose in all populations (P < 0.05) and reduced mRNA of insulin receptor (IR) and GLUT 2 and 3 in liver (P < 0.05). There was heterosis for mRNA, most notably NPYR1 (-78%) and NPYR5 (-81%) in fat and GLUT2 (-70%) in liver. Results suggest that NPY and GLUTs are associated with differences in energy homeostasis in LWS and HWS. Reduced GLUT and IR mRNA after insulin injection suggest a compensatory mechanism to prevent further hypoglycemia.
Asunto(s)
Apetito/genética , Peso Corporal/genética , Proteínas Facilitadoras del Transporte de la Glucosa/metabolismo , Insulina/farmacología , ARN Mensajero/metabolismo , Receptores de Neuropéptido/metabolismo , Grasa Abdominal/metabolismo , Animales , Glucemia/metabolismo , Pollos , Cruzamientos Genéticos , Femenino , Factores de Transcripción Forkhead/genética , Factores de Transcripción Forkhead/metabolismo , Vigor Híbrido , Hipotálamo/metabolismo , Hígado/metabolismo , Músculo Esquelético/metabolismo , Neuropéptido Y/genética , Neuropéptido Y/metabolismo , Especificidad de Órganos , ARN Mensajero/genética , Receptor de Insulina/genética , Receptor de Insulina/metabolismo , Receptores de Neuropéptido/genética , Especificidad de la EspecieRESUMEN
As a member of the interleukin (IL)-10 family, IL-22 is an important mediator in modulating tissue responses during inflammation. Through activation of STAT3-signaling cascades, IL-22 induces proliferative and anti-apoptotic pathways, as well as antimicrobial peptides (AMPs), that help prevent tissue damage and aid in its repair. This study reports the cloning and expression of recombinant chicken IL-22 (rChIL-22) and its soluble receptor, rChIL22BP, and characterization of biological effects of rChIL-22 during inflammatory responses. Similar to observations with mammalian IL-22, purified rChIL-22 had no effect on either peripheral blood mononuclear cells (PBMCs) or lymphocytes. This was due to the low expression of the receptor ChIL22RA1 chain compared to ChIL10RB chain. rChIL-22 alone did not affect chicken embryo kidney cells (CEKCs); however, co-stimulation of CEKCs with LPS and rChIL-22 enhanced the production of pro-inflammatory cytokines, chemokines and AMPs. Furthermore, rChIL-22 alone stimulated and induced acute phase reactants in chicken embryo liver cells (CELCs). These effects of rChIL-22 were abolished by pre-incubation of rChIL-22 with rChIL22BP. Together, this study indicates an important role of ChIL-22 on epithelial cells and hepatocytes during inflammation.
Asunto(s)
Células Epiteliales/inmunología , Hepatocitos/inmunología , Inflamación/inmunología , Interleucinas/genética , Interleucinas/inmunología , Receptores de Interleucina/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Línea Celular , Proliferación Celular , Embrión de Pollo , Pollos/inmunología , Fibroblastos , Interleucinas/química , Interleucinas/metabolismo , Leucocitos Mononucleares/inmunología , Datos de Secuencia Molecular , Receptores de Interleucina/química , Receptores de Interleucina/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/metabolismo , Análisis de Secuencia de ADN , Distribución Tisular , Interleucina-22RESUMEN
Intestinal colonization of avian species by Eimeria parasites results in the enteric disease, coccidiosis. A study was carried out to assess the immunologic effects of Eimeria praecox infection on the gut of infected chickens. In Experiment 1, birds were orally gavaged with 50,000 E. praecox oocysts; in Experiment 2, an infection dosage of 500,000 E. praecox oocysts was used. Duodenal and jejunal intestinal sections were sampled consecutively on days 1-7 post-infection. Intestinal expression of innate immune gene transcripts was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). Analysis of relative gene expression in Experiment 1 revealed an increase (P<0.05) in duodenal Toll-like receptor (TLR)3 expression on days 4 and 6 post-infection. TLR15 expression was significantly decreased in the duodenum of infected birds on day 2, and significantly increased on day 6 post-infection. In Experiment 2, TLR3 was significantly downregulated in the duodenum on day 7 post-infection; however, no significant results were observed in terms of TLR15 expression. TLR4 also exhibited decreased expression (P<0.05) on day 7 post-infection in both intestinal sections. Regarding antimicrobial peptide expression; in the first experiment, expression of liver-expressed antimicrobial peptide-2 (LEAP-2) in infected birds was significantly decreased in the duodenum on days 3 and 4, and in the jejunum on day 4. Similarly, Experiment 2 resulted in depression of LEAP-2 (P<0.05) on days 3-5 in the duodenum. In Experiment 1, cathelicidin antimicrobial peptide (CATHL3) was downregulated (P<0.05) in the jejunum of infected chickens on day 3 post-infection; however, CATHL3 results were non-significant in Experiment 2. Based on the differing results observed in each experiment, it was concluded that both TLR and antimicrobial peptide expression, and thus immunity may be dependent on infection load.