RESUMEN
Barramundi (Lates calcarifer) are a highly valued aquaculture species, and, as obligate carnivores, they have a demonstrated preference for dietary protein over lipid or starch to fuel energetic growth demands. In order to investigate how carnivorous fish regulate nutritional cues, we examined the metabolic effects of feeding two isoenergetic diets that contained different proportions of digestible protein or starch energy. Fish fed a high proportion of dietary starch energy had a higher proportion of liver SFA, but showed no change in plasma glucose levels, and few changes in the expression of genes regulating key hepatic metabolic pathways. Decreased activation of the mammalian target of rapamycin growth signalling cascade was consistent with decreased growth performance values. The fractional synthetic rate (lipogenesis), measured by TAG 2H-enrichment using 2H NMR, was significantly higher in barramundi fed with the starch diet compared with the protein diet (0·6 (se 0·1) v. 0·4 (se 0·1) % per d, respectively). Hepatic TAG-bound glycerol synthetic rates were much higher than other closely related fish such as sea bass, but were not significantly different (starch, 2·8 (se 0·3) v. protein, 3·4 (se 0·3) % per d), highlighting the role of glycerol as a metabolic intermediary and high TAG-FA cycling in barramundi. Overall, dietary starch significantly increased hepatic TAG through increased lipogenesis. Compared with other fish, barramundi possess a unique mechanism to metabolise dietary carbohydrates and this knowledge may define ways to improve performance of advanced formulated feeds.
Asunto(s)
Lubina/metabolismo , Dieta/veterinaria , Lipogénesis/efectos de los fármacos , Almidón/farmacología , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Proteínas en la Dieta/farmacología , Hígado/metabolismoRESUMEN
Based on the concept of nutritional programming in mammals, we tested whether an acute hyperglucidic-hypoproteic stimulus during first feeding could induce long-term changes in nutrient metabolism in rainbow trout. Trout alevins received during the five first days of exogenous feeding either a hyperglucidic (40% gelatinized starch + 20% glucose) and hypoproteic (20%) diet (VLP diet) or a high-protein (60%) glucose-free diet (HP diet, control). Following a common 105-day period on a commercial diet, both groups were then challenged (65 days) with a carbohydrate-rich diet (28%). Short- and long-term effects of the early stimuli were evaluated in terms of metabolic marker gene expressions and intestinal microbiota as initial gut colonisation is essential for regulating the development of the digestive system. In whole alevins (short term), diet VLP relative to HP rapidly increased gene expressions of glycolytic enzymes, while those involved in gluconeogenesis and amino acid catabolism decreased. However, none of these genes showed persistent molecular adaptation in the liver of challenged juveniles (long term). By contrast, muscle of challenged juveniles subjected previously to the VLP stimulus displayed downregulated expression of markers of glycolysis and glucose transport (not seen in the short term). These fish also had higher plasma glucose (9 h postprandial), suggesting impaired glucose homeostasis induced by the early stimulus. The early stimulus did not modify the expression of the analysed metabolism-related microRNAs, but had short- and long-term effects on intestinal fungi (not bacteria) profiles. In summary, our data show that a short hyperglucidic-hypoproteic stimulus during early life may have a long-term influence on muscle glucose metabolism and intestinal microbiota in trout.
Asunto(s)
Metabolismo de los Hidratos de Carbono/fisiología , Carbohidratos de la Dieta/metabolismo , Proteínas en la Dieta/metabolismo , Intestinos/microbiología , Microbiota , Oncorhynchus mykiss/crecimiento & desarrollo , Oncorhynchus mykiss/metabolismo , Animales , Glucemia , Dieta , Expresión Génica/fisiología , Gluconeogénesis/genética , Glucólisis/genética , Homeostasis/efectos de los fármacos , Hígado/metabolismo , Músculos/metabolismoRESUMEN
Carnivorous fish species such as the rainbow trout (Oncorhynchus mykiss) are considered to be "glucose intolerant" because of the prolonged hyperglycemia experienced after intake of a carbohydrate-enriched meal. In the present study, we use this species to study glucose homeostasis in fish chronically infused with the hypoglycemic agents, insulin, and metformin, and fed with a high proportion of carbohydrates (30%). We analyzed liver, skeletal muscle, and white adipose tissue (WAT), which are insulin- and metformin-specific targets at both the biochemical and molecular levels. Trout infused with the combination of insulin and metformin can effectively utilize dietary glucose at the liver, resulting in lowered glycemia, increased insulin sensitivity, and glucose storage capacity, combined with reduced glucose output. However, in both WAT and skeletal muscle, we observed decreased insulin sensitivity with the combined insulin + metformin treatment, resulting in the absence of changes at the metabolic level in the skeletal muscle and an increased potential for glucose uptake and storage in the WAT. Thus, the poor utilization by rainbow trout of a diet with a high proportion of carbohydrate can at least be partially improved by a combined treatment with insulin and metformin, and the glucose intolerance observed in this species could be, in part, due to some of the downstream components of the insulin and metformin signaling pathways. However, the predominant effects of metformin treatment on the action of insulin in these three tissues thought to be involved in glucose homeostasis remain exclusive in this species.
Asunto(s)
Carbohidratos de la Dieta/farmacología , Glucosa/metabolismo , Homeostasis/efectos de los fármacos , Insulina/farmacología , Metformina/farmacología , Oncorhynchus mykiss/metabolismo , Tejido Adiposo Blanco/metabolismo , Administración Oral , Animales , Glucemia/metabolismo , Carbohidratos de la Dieta/administración & dosificación , Interacciones Farmacológicas , Homeostasis/fisiología , Hipoglucemiantes/farmacología , Bombas de Infusión , Insulina/administración & dosificación , Hígado/metabolismo , Metformina/administración & dosificación , Modelos Animales , Músculo Esquelético/metabolismoRESUMEN
The origin for the poor glucose utilization in carnivorous fish species fed high carbohydrate diets remains under debate. In the present study, we have fed rainbow trout a diet containing 30% carbohydrate for 1 or 5 days. In both cases, fish were implanted with mini-osmotic pumps releasing 0.7 i.u. kg(-1) day(-1) bovine insulin, and mRNA transcripts and the protein phosphorylation status of proteins controlling glycemia and glucose-related metabolism were studied in fish killed 6 h after the last meal. We demonstrate that when the exposure occurs over a short term (30 h), insulin exerts beneficial actions on trout glucose homeostasis, including a lowered glycemia and increased hepatic lipogenic and glycogenic potentials. However, when trout were fed for 5 days, these beneficial actions of insulin infusion were no longer observed. Thus, the increased lipogenic potential observed after one single meal was not present, and this together with the increased glycogenesis and the decreased glucose exported to the blood from the liver explains the lack of hypoglycemic action of insulin. The fact that insulin improved glucose homeostasis when administrated over a short time period implies that endogenous insulin secretion is inadequate in trout to deal with this amount of dietary carbohydrates. Moreover, the fact that a longer exposure to insulin resulted in a reduced response indicates that the rainbow trout is sensitive to insulin, re-enforcing the hypothesis that the hyperglycemia observed following a high carbohydrate meal is an insulin secretion issue rather an insulin action issue.
Asunto(s)
Carbohidratos de la Dieta/farmacología , Proteínas de Peces/metabolismo , Glucosa/metabolismo , Insulina/farmacología , Oncorhynchus mykiss/fisiología , Tejido Adiposo Blanco/metabolismo , Animales , Proteínas de Peces/genética , Homeostasis/efectos de los fármacos , Bombas de Infusión Implantables , Hígado/metabolismo , Músculo Esquelético/metabolismo , Oncorhynchus mykiss/metabolismo , Fosforilación , ARN Mensajero/metabolismoRESUMEN
Utilizing rainbow trout (Oncorhynchus mykiss) as a known model of a "glucose-intolerant" and poor dietary glucose user, we assessed glucose utilization in fish chronically receiving two molecules able to improve glucose homeostasis: insulin and metformin. Our objectives were to assess the ability of rainbow trout to deal with a glucose load and to improve glucose utilization in fish receiving a chronic administration of insulin plus metformin treatments. Fish received (implanted miniosmotic pumps) saline, insulin, metformin, and insulin plus metformin solution for 4 days and then were subjected to a glucose challenge (intraperitoneal injection) to study glucose homeostasis, analyzing plasma glycemia, mRNA levels of glucose metabolism-related proteins, insulin signaling, and glycogen levels in liver and muscle. Control fish received a saline pump implantation and saline intraperitoneal injection. We found no evidence that the "glucose intolerance" in this species could be linked to any of the molecular markers of metabolism in the tissues analyzed. By contrast, very interestingly, we show for the first time, that metformin is not only unable to improve glucose homeostasis in trout, but, in fact, its counteracts the effects of insulin, creating an "insulin resistance," especially in the muscle. These results make trout an attractive original model to study both insulin and metformin effect on biological systems.
Asunto(s)
Glucemia/efectos de los fármacos , Intolerancia a la Glucosa/tratamiento farmacológico , Hipoglucemiantes/farmacología , Resistencia a la Insulina , Insulina/farmacología , Metformina/farmacología , Oncorhynchus mykiss/metabolismo , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Conducta Alimentaria , Regulación de la Expresión Génica/efectos de los fármacos , Gluconeogénesis/efectos de los fármacos , Gluconeogénesis/genética , Glucosa/administración & dosificación , Intolerancia a la Glucosa/genética , Intolerancia a la Glucosa/metabolismo , Intolerancia a la Glucosa/fisiopatología , Proteínas Facilitadoras del Transporte de la Glucosa/genética , Proteínas Facilitadoras del Transporte de la Glucosa/metabolismo , Glucólisis/efectos de los fármacos , Glucólisis/genética , Homeostasis , Hipoglucemiantes/administración & dosificación , Bombas de Infusión Implantables , Inyecciones Intraperitoneales , Insulina/administración & dosificación , Resistencia a la Insulina/genética , Lipogénesis/efectos de los fármacos , Lipogénesis/genética , Hígado/efectos de los fármacos , Hígado/metabolismo , Glucógeno Hepático/metabolismo , Metformina/administración & dosificación , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/metabolismo , Oncorhynchus mykiss/genética , ARN Mensajero/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Factores de TiempoRESUMEN
Carnivorous fish are poor users of dietary carbohydrates and are considered to be glucose intolerant. In this context, we have tested, for the first time in rainbow trout, metformin, a common anti-diabetic drug, known to modify muscle and liver metabolism and to control hyperglycemia in mammals. In the present study, juvenile trout were fed with very high levels of carbohydrates (30% of the diet) for this species during 10 days followed by feeding with pellets supplemented with metformin (0.25% of the diet) for three additional days. Dietary metformin led to a significant reduction in postprandial glycemia in trout, demonstrating unambiguously the hypoglycemic effect of this drug. No effect of metformin was detected on mRNA levels for glucose transporter type 4 (GLUT4), or enzymes involved in glycolysis, mitochondrial energy metabolism, or on glycogen level in the white muscle. Expected inhibition of hepatic gluconeogenic (glucose-6-phosphatase, fructose-1,6-bisphosphatase, and phosphoenolpyruvate carboxykinase) mRNA levels was not found, showing instead paradoxically higher mRNA levels for these genes after drug treatment. Finally, metformin treatment was associated with higher mRNA levels and activities for lipogenic enzymes (fatty acid synthase and glucose-6-phosphate dehydrogenase). Overall, this study strongly supports that the induction of hepatic lipogenesis by dietary glucose may permit a more efficient control of postprandial glycemia in carnivorous fish fed with high carbohydrate diets.
Asunto(s)
Glucemia/efectos de los fármacos , Carbohidratos de la Dieta/metabolismo , Metabolismo Energético/efectos de los fármacos , Hipoglucemiantes/farmacología , Lipogénesis/efectos de los fármacos , Hígado/efectos de los fármacos , Metformina/farmacología , Oncorhynchus mykiss/metabolismo , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Glucemia/metabolismo , Carbohidratos de la Dieta/administración & dosificación , Metabolismo Energético/genética , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Homeostasis , Lipogénesis/genética , Hígado/enzimología , Hígado/metabolismo , Periodo Posprandial , ARN Mensajero/metabolismo , Factores de TiempoRESUMEN
The rapid development of aquaculture production throughout the world over the past few decades has led to the emergence of new scientific challenges to improve fish nutrition. The diet formulations used for farmed fish have been largely modified in the past few years. However, bottlenecks still exist in being able to suppress totally marine resources (fish meal and fish oil) in diets without negatively affecting growth performance and flesh quality. A better understanding of fish metabolism and its regulation by nutrients is thus mandatory. In this review, we discuss four fields of research that are highly important for improving fish nutrition in the future: ( a) fish genome complexity and subsequent consequences for metabolism, ( b) microRNAs (miRNAs) as new actors in regulation of fish metabolism, ( c) the role of autophagy in regulation of fish metabolism, and ( d) the nutritional programming of metabolism linked to the early life of fish.
Asunto(s)
Autofagia , Aceites de Pescado/metabolismo , Peces/metabolismo , Genoma/genética , Genómica , MicroARNs/genética , Animales , Acuicultura , Dieta/veterinaria , Peces/genética , Estado NutricionalRESUMEN
Genetic selection has significantly improved the muscle development of fast-growing broiler chickens in the last 50 yr. However, improvement in muscle growth has coincided with relatively poor development of visceral systems, resulting in impaired ability to cope with high environmental temperatures. The aim of this study was to elucidate the effects of thermal manipulation (TM) during different periods of embryogenesis on chick hatchability, BW and thermoregulation upon hatching, on their ability to cope with thermal challenge at 42 d of age, and on carcass and breast meat traits. Control embryos were incubated at 37.8 degrees C. The TM embryos were incubated at 37.8 degrees C and treated for 3 h at 39.5 degrees C on the following days of embryogenesis: E8 to E10 [early (EA)], E16 to E18 [late (LA)], and both E8 to E10 and E16 to E18 (EA-LA). Body weight and body temperature (T(b)) were measured at hatching and throughout the growth period as well as during exposure of 42-d-old chickens to a thermal challenge at 35 degrees C for 6 h. The LA and EA chicks exhibited significantly lower T(b) than control chicks (37.9 vs. 38.2 degrees C) at hatching, but during the growth period, differences in T(b) between treated and control chicks decreased with age. Significant hyperthermia (over 44 degrees C) was monitored in all groups during the thermal challenge, but mortality was higher in treated than in control chickens. No effect of treatments on BW was found during the entire growth period. However, breast yield was higher in LA chickens than in controls at slaughter. The EA and EA-LA treatments slightly decreased the ultimate pH of breast meat, whereas the LA treatment had no effect. In conclusion, none of the TM conditions tested in the present study were able to improve long-term thermotolerance in chickens. Late treatment favored breast muscle growth without affecting ultimate pH and drip loss of breast meat.
Asunto(s)
Adaptación Fisiológica/fisiología , Regulación de la Temperatura Corporal/fisiología , Pollos/fisiología , Músculo Esquelético/fisiología , Temperatura , Animales , Composición Corporal , Peso Corporal/fisiología , Embrión de Pollo , Calor , Carne/normas , Factores de TiempoRESUMEN
We investigated whether a change in peripheral glucose homeostasis, a local change in the insulin-related ovarian regulatory system, or both occurred in ad libitum-fed broiler breeder hens compared with feed-restricted counterparts. Feed-restricted (R, from 5 to 16 wk of age) and ad libitum-fed (A) hens from a standard commercial line (S) and an experimental dwarf genotype (E) were studied. Basal and stimulated plasma insulin and glucose concentrations were measured during the prebreeding and laying periods. In the basal state (after 16 h fasting) plasma glucose concentrations were significantly lower in SA chickens (-5% at 17 wk, -7.5% at 32 wk) compared with EA, SR, and ER chickens, with no difference in plasma insulin concentrations (n = 16). In 17-wk-old SA birds, 30 min after oral glucose loading, plasma glucose concentrations increased significantly compared with the basal state and were also significantly lower as compared with SR but did not differ significantly from EA and ER. Plasma insulin concentrations did not differ significantly between genotypes or regimens (n = 16). A potential modification of intracellular mediators involved in the regulation of cell growth and survival in small follicles that were overrecruited in SA compared with SR was also investigated in SA and SR hens at 32 wk. There was no effect of food restriction in phospho-Akt, Akt, phospho-ERK, and phospho-S6 in the small white ovarian follicles (n = 6) in the basal state and after 30 min of refeeding. In conclusion, the present study does not demonstrate any evidence of glucose intolerance during the prebreeding period, specific change in the ovarian small follicle insulin signalling pathway, or both, in laying broiler breeders fed ad libitum compared with feed-restricted hens.
Asunto(s)
Glucemia/metabolismo , Pollos/fisiología , Dieta , Insulina/farmacología , Ovario/efectos de los fármacos , Animales , Glucemia/análisis , Cruzamiento , Pollos/genética , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Ayuno , Femenino , Alimentos , Genotipo , Prueba de Tolerancia a la Glucosa/veterinaria , Homeostasis , Insulina/sangre , Ovario/enzimología , Ovario/fisiología , Oviposición , Fosforilación , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteínas Quinasas S6 Ribosómicas/metabolismoRESUMEN
Ad libitum feeding reduces livability and reproductive fitness in broiler breeder hens. Two genotypes, a standard (S) and an experimental dwarf broiler breeder (E), were fed ad libitum (SA and EA, respectively), restricted at 55% of ad libitum feed intake (intermediate restriction) from 6 to 15 wk of age (SI and EI, respectively), or restricted (SR and ER, respectively) to match a standard growth curve with a diluted mash feed (2,400 kcal/kg). The experiment was repeated at 2 locations (experiment 1 = 672 hens in pens from 0 to 40 wk; experiment 2 = 420 hens in pens and cages from 0 to 53 wk). Feed restriction reduced adult BW by 20% compared with ad libitum feeding, delayed sexual maturity by 2 to 4 wk, and improved livability. Hens fed the intermediate diet immediately compensated after 15 wk of age to reach BW, sexual maturity, and livability close to those of ad libitum-fed hens. The E genotype exhibited better tolerance to ad libitum feeding than the S genotype in all measured aspects. Average laying rate during the first 24 wk of lay was 66.4, 77.4, 69.9, 47.2, 57.9, and 72.4% for EA, EI, ER, SA, SI, and SR respectively in experiment 2. Egg abnormalities (double yolk, shell problems) decreased after the peak of lay but remained consistently higher for S compared with E, and for ad libitum and intermediate diets compared with the restricted diet. Yolk deposition rate was measured by a double dye technique. Duration of yolk rapid growth was 8.8 d in E and 9.3 d in S hens (P < 0.001), but this difference did not explain the observed variations in laying rate. The potential to increase feed allowances even with a diluted diet in broiler breeder hens requires adapted genotypes.
Asunto(s)
Pollos/genética , Dieta/veterinaria , Envejecimiento , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Yema de Huevo/fisiología , Femenino , Privación de Alimentos , Genotipo , Oviposición , Óvulo/fisiología , Aumento de PesoRESUMEN
In a previous study we showed the existence of GH in the ovine placenta. We now supplement the information available on placental GH and describe the presence and distribution of GH receptor (GH-R) messenger RNA (mRNA) in uterine, fetal, and placental tissues during early pregnancy. GH mRNA was not detected in the placenta before day 27 (d27). Its expression peaked between d40 and d45 and fell after d55. GH mRNA was localized in the trophectoderm and syncytium. During the d35-d50 period, concentrations of GH in the maternal circulation were not increased. In umbilical blood, however, GH was detected from d35 and was presumed to be of placental origin, because GH mRNA was not detected in the fetal pituitary gland on d40. We report on GH-R mRNA expression in the placenta between d20-d120. The relative abundance of GH-R transcripts increased significantly between d25-d43. In the endometrium, GH-R mRNA was detected from d8-d120 of pregnancy and from d4-d16 of the cycle. GH-R mRNA was localized in the trophectoderm, fetal mesoderm, and maternal uterine stroma. In the fetal liver, GH-R mRNA was first detectable on d35. The results of this study indicate that between d35-d50 of pregnancy, the endometrium, placenta, and fetus are all potential targets for the placental GH.
Asunto(s)
Feto/metabolismo , Expresión Génica , Hormona del Crecimiento/genética , Placenta/metabolismo , Receptores de Somatotropina/genética , Alantoides/metabolismo , Líquido Amniótico/química , Animales , Endometrio/química , Femenino , Sangre Fetal/química , Edad Gestacional , Hormona del Crecimiento/análisis , Hormona del Crecimiento/sangre , Hígado/química , Hígado/embriología , Hipófisis/química , Hipófisis/embriología , Placenta/química , Embarazo , ARN Mensajero/análisis , Ovinos , Trofoblastos/química , Útero/químicaRESUMEN
In this study, we have investigated the expression of the prolactin receptor gene in ovine endometrium during oestrus cycle and pregnancy. Using reverse transcription-PCR analysis, we provided evidence that the prolactin receptor gene is specifically transcribed in this tissue. As shown by Northern blot analysis, the level of the prolactin receptor transcripts increased dramatically during late pregnancy. In situ hybridization experiments revealed that prolactin receptor mRNA was specifically expressed in the glandular compartment and confirmed the dramatic increase of its expression that occurs at the end of pregnancy. Taken together, these findings are consistent with a putative role of prolactin and/or related molecules in the regulation of the proliferation of the glandular compartment and/or in the control of the secretory activity of the endometrium.
Asunto(s)
Endometrio/metabolismo , Evolución Molecular , Receptores de Prolactina/genética , Animales , Northern Blotting , Endometrio/patología , Femenino , Hibridación in Situ , Reacción en Cadena de la Polimerasa , Embarazo , ARN Mensajero , ADN Polimerasa Dirigida por ARN , Ovinos , ÚteroRESUMEN
The acute effect of leptin on the regulation of food intake was investigated in layer and broiler chickens. In an initial study, we observed that a single intraperitoneal injection of recombinant chicken leptin (1 mg/kg BW) dramatically reduced (38%) food intake in 56-day-old layer chickens, more moderately reduced (15%) food intake in 9-day-old layer chicks, and had no significant effect in 9-day-old broiler chicks. In a subsequent study, body weight and plasma concentrations of leptin were measured weekly in layer and broiler chicks from day 1 to 35 of age and brain leptin receptor and neuropeptide Y (NPY) mRNA expression were analyzed at 1, 9, and 35 days of age. At day 1 of age, peripheral concentrations of leptin were significantly greater in layer than broiler chicks. Subsequently, despite increases in body weight and differences in growth rates between layer and broiler chicks from day 8 to day 35 of age, peripheral concentrations of leptin were constant and similar in both genotypes. Leptin receptor and NPY mRNA were expressed in brain from day 1 in chicks of both genotypes and increased significantly to day 35 of age. These observations provide evidence that the inhibitory effect of leptin on the regulation of food intake in growing chicks is an age dependent process. Furthermore, acquisition of the anorectic effect of leptin is likely to be associated with greater expression of the leptin receptor and NPY mRNAs than to changes in blood levels of leptin. Finally, this study provides evidence that chickens selected for high growth rates may be less sensitive or responsive to peripheral concentrations of leptin than chickens with low growth rates (layers), suggesting that the faster growth of broiler chicks may be related to a lessened responsiveness to anorexigenic factors.
Asunto(s)
Conducta Animal , Ingestión de Alimentos/fisiología , Conducta Alimentaria/fisiología , Leptina/fisiología , Factores de Edad , Animales , Encéfalo/metabolismo , Pollos , Leptina/sangre , Masculino , Neuropéptido Y/genética , Neuropéptido Y/metabolismo , ARN Mensajero/análisis , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/metabolismo , Receptores de Leptina , Especificidad de la EspecieRESUMEN
In order to examine the hormonal regulation of the prolactin-receptor (PRL-R) gene expression during mammary gland development, ewes were treated to induce lactation via an estrogen-progesterone-hydrocortisone and ovine growth hormone treatment. In situ hybridization analysis was used and revealed that sex steroids increased PRL-R mRNA levels in the mammary gland. Using RNase protection assay we showed that the estradiol + progesterone treatment increased both the levels of the long and the short forms of PRL-R mRNA. Addition of hydrocortisone increased the level of alphaS1-casein transcripts and the level of the ratio of the long to the short form of the PRL-R mRNA. This ratio can be further enhanced by addition of ovine growth hormone to the latter treatment. This suggests a role of hydrocortisone and ovine growth hormone in the alternative splicing that leads to the preferential expression of the long form of the PRL-R mRNA. In conclusion, the present experiments suggest that estrogen, progesterone and hydrocortisone are the major regulators of the PRL-R gene expression during pregnancy and prepare the mammary gland for its differentiation.
Asunto(s)
Regulación del Desarrollo de la Expresión Génica , Lactancia , Glándulas Mamarias Animales/fisiología , Receptores de Prolactina/genética , Ovinos/fisiología , Empalme Alternativo/fisiología , Animales , Northern Blotting/veterinaria , Cartilla de ADN/química , Electroforesis en Gel de Agar , Electroforesis en Gel de Poliacrilamida/veterinaria , Estrógenos/fisiología , Femenino , Hormona del Crecimiento/fisiología , Hidrocortisona/fisiología , Procesamiento de Imagen Asistido por Computador , Hibridación in Situ/veterinaria , Glándulas Mamarias Animales/crecimiento & desarrollo , Progesterona/fisiología , ARN Mensajero/biosíntesis , ARN Mensajero/química , ARN Mensajero/aislamiento & purificación , Receptores de Prolactina/análisis , Receptores de Prolactina/fisiología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Ribonucleasas/química , Ovinos/crecimiento & desarrolloRESUMEN
Chicken leptin cDNA shows a high homology to mammalian homologous, with an expression localized in the liver and adipose tissue. It is noteworthy, that the hepatic expression is most likely associated with the primary role that this organ plays in lipogenic activity in avian species. As in mammals, chicken leptin expression is regulated by hormonal and nutritional status. This regulation is tissue-specific and with a high sensitivity in the liver compared to adipose tissue. The blood leptin levels are regulated by the nutritional state with high levels in the fed state compared to the fasted state. The recombinant chicken leptin markedly inhibits food intake as reported in mammals, suggesting the presence of an hypothalamic leptin receptor. The chicken leptin receptor has been identified and all functional motifs are highly conserved compared to mammalian homologous. Chicken leptin receptor is expressed in the hypothalamus but also in other tissues such as pancreas, where leptin inhibits insulin secretion and thus may have a key role in regulating nutrient utilization in this species.
Asunto(s)
Pollos/fisiología , Leptina/fisiología , Receptores de Superficie Celular , Tejido Adiposo/metabolismo , Tejido Adiposo/fisiología , Animales , Proteínas Portadoras/metabolismo , Proteínas Portadoras/fisiología , Pollos/metabolismo , Ingestión de Alimentos , Regulación de la Expresión Génica/fisiología , Leptina/metabolismo , Hígado/metabolismo , Hígado/fisiología , Receptores de LeptinaRESUMEN
In chickens, leptin is expressed mainly in the liver, where its receptor gene expression has also been reported, and in adipose tissue. In view of the key role played by the liver in lipogenesis in avian species, the hepatic expression of leptin may have physiological significance. In this study, we showed that leptin is constitutively expressed and secreted in a chicken-derived hepatoma cell line (LMH). Although insulin regulates leptin expression in vivo, incubation of LMH cells in the presence of 100 nM insulin for 24 or 48 h had no effect on leptin expression or its secretion in the culture medium. In addition, we developed a specific chicken leptin receptor real-time reverse transcription (RT)-PCR, and downregulation of leptin receptor gene expression by homologous and heterologous signals was demonstrated, as relative leptin receptor mRNA levels were significantly decreased after exposure of LMH cells to recombinant chicken leptin or porcine insulin. In conclusion, our results indicate that leptin is probably able to desensitize its own response in the chicken liver. Finally, the ability of insulin and leptin to regulate chicken leptin receptor gene expression suggests a direct role of leptin in the control of hepatic metabolism.
Asunto(s)
Carcinoma Hepatocelular/veterinaria , Insulina/farmacología , Leptina/farmacología , Neoplasias Hepáticas/veterinaria , Enfermedades de las Aves de Corral/metabolismo , Receptores de Superficie Celular/genética , Animales , Western Blotting , Carcinoma Hepatocelular/metabolismo , Dexametasona/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Leptina/genética , Leptina/metabolismo , Neoplasias Hepáticas/metabolismo , ARN Mensajero/análisis , Receptores de Leptina , Proteínas Recombinantes , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Triyodotironina/farmacología , Células Tumorales CultivadasRESUMEN
Although the metabolic actions of insulin in fish have been investigated widely in the past several years, lipid metabolism has received little attention, especially in tissues like the liver or white muscle. In the present study, rainbow trout received insulin treatments both acutely (intraperitoneal injection) and chronically (through mino-osmotic pumps) to elucidate hormone metabolic actions at molecular levels on the 2 main insulin target tissues in trout, namely, liver and muscle. Plasma and free fatty acid concentrations in plasma, as well as mRNA measurements of some key enzymes involved in lipid metabolism, were assessed in these tissues after 6h and 4 d of acute and chronic insulin treatments, respectively. Our results showed that although fish received the same final total amount of hormone in both treatments, the actions of insulin on lipid metabolism were both time and tissue dependent. After the acute insulin treatment, the main anabolic role of insulin was reflected in decreased plasma free fatty acid concentrations linked to enhanced hepatic lipogenesis. We also found that insulin increased the mRNA levels of enzymes involved in lipid oxidation, perhaps to counteract insulin-induced hypoglycemia. In contrast, our data show that after chronic insulin treatment, liver and muscle exhibit different metabolic strategies: whereas in the liver chronic insulin-induced hypoglycemia may stimulate lipolytic processes to spare glucose stores, the muscle responds directly to the anabolic hormone action by increasing its lipogenic capacity and by inhibiting pathways of lipid oxidation.
Asunto(s)
Insulina/administración & dosificación , Metabolismo de los Lípidos , Hígado/metabolismo , Músculos/metabolismo , Oncorhynchus mykiss/metabolismo , Animales , Glucemia/análisis , Glucemia/efectos de los fármacos , Ácido Graso Sintasas/genética , Ácidos Grasos no Esterificados/sangre , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Glucosafosfato Deshidrogenasa/genética , Metabolismo de los Lípidos/efectos de los fármacos , Metabolismo de los Lípidos/genética , Peroxidación de Lípido/efectos de los fármacos , Lipólisis/efectos de los fármacos , Hígado/efectos de los fármacos , Músculos/efectos de los fármacos , Oncorhynchus mykiss/genética , Especificidad de Órganos , Reacción en Cadena de la Polimerasa/veterinaria , ARN Mensajero/análisisRESUMEN
To explore the mechanisms leading to excessive adiposity in chicken, we investigated the regulation of fatty acid oxidation depending on genotype-related body fatness and diet composition. mRNA expression and/or activity of proteins involved in mitochondrial energy metabolism were measured in liver and gastrocnemius muscle of genetically lean or fat chickens reared on a low-fat/high-protein diet or an isoenergetic high-fat/low-protein diet (HF/LP). Muscle expressions of the muscle isoform of carnitine-palmitoyltransferase 1 (M-CPT1) and PPARbeta/delta were higher in fat than in lean chickens. This was also observed in liver, although only with the HF/LP diet for M-CPT1. This could stimulate mitochondrial fatty acid oxidation in fat chickens. Up-regulations of liver and muscle CPT-1 hepatic isoform, and muscle cytochrome-c-oxidase mRNA expressions, and of beta-hydroxyacyl-CoA-dehydrogenase activities suggest higher fatty acid utilization with the HF/LP diet. PPARbeta/delta and PGC-1alpha could control fatty acid oxidation in muscle and liver, respectively. Regulation of avian uncoupling protein (avUCP) mRNA was tissue-dependent. Predominantly expressed in muscle, it was stimulated in fat and in HF/LP-fed chickens, where it could be associated to the special need in muscle anti-oxidant pathways of fatter animals. In liver it was lower in fat than in lean chickens, and its potential function remains to be clarified.
Asunto(s)
Pollos/metabolismo , Dieta , Metabolismo Energético/fisiología , Ácidos Grasos/metabolismo , Oxidación-Reducción , Animales , Carnitina O-Palmitoiltransferasa/genética , Carnitina O-Palmitoiltransferasa/metabolismo , Pollos/genética , Dieta con Restricción de Grasas , Dieta con Restricción de Proteínas , Metabolismo Energético/genética , Genotipo , Metabolismo de los Lípidos/fisiología , Masculino , Mitocondrias Hepáticas/fisiología , Proteínas Mitocondriales , Músculo Esquelético/fisiología , ARN Mensajero/metabolismoRESUMEN
In this study, we have analyzed the developmental expression of the prolactin receptor (PRL-R) gene in the ewe mammary gland during pregnancy and lactation. Using Northern and slot-blot analysis and in situ hybridization, we showed that the level of PRL-R mRNA in mammary epithelial cells increased during the second half of pregnancy, decreased at the end of pregnancy, and remained relatively stable during lactation with a level above that observed at the beginning of pregnancy. As shown by RNase protection assay, the ratio of the long to the short form of the PRL-R mRNA was always above 1. This ratio increased between Day 70 of pregnancy and term and decreased progressively during lactation. The high level of PRL-R mRNA before the induction of alphaS1-casein gene expression suggests that PRL may be involved in the growth and development of the mammary gland. More precisely, the increase of the ratio of the long to the short form of the PRL-R during lactogenesis suggests that the latter form may have a dominant negative action in the activation of milk protein gene transcription. Thus the long/short-form ratio of the PRL-R may play a key role in the shift between growth and differentiation of the mammary gland.
Asunto(s)
Regulación del Desarrollo de la Expresión Génica/fisiología , Lactancia/metabolismo , Glándulas Mamarias Animales/crecimiento & desarrollo , Glándulas Mamarias Animales/metabolismo , Preñez/metabolismo , ARN Mensajero/biosíntesis , Receptores de Prolactina/biosíntesis , Animales , Femenino , Regulación del Desarrollo de la Expresión Génica/genética , Immunoblotting , Hibridación in Situ , Lactancia/genética , Embarazo , Preñez/genética , ARN Mensajero/genética , Receptores de Prolactina/genética , Ribonucleasas/metabolismo , OvinosRESUMEN
Thirty-four-week-old laying hens received injections of recombinant chicken leptin to assess the role of leptin in avian ovarian function. In the first experiment, the hens (n=60) were divided into three groups: (i). fed ad libitum; (ii). fasted; and (iii). fasted + leptin. Hens were fasted for 5 days and those treated with leptin received 250 microg leptin kg-1 body weight twice a day, i.p. In the second experiment, the hens (n=72) were divided into four groups: (i). fed ad libitum; (ii). fasted; (iii). fasted + leptin given only during fasting (5 days); or (iv). fasted and leptin given during both fasting and 5 days of re-feeding (10 days). LH was measured in blood plasma, and progesterone and oestradiol were measured in blood plasma and the ovary by radioimmunoassay. Apoptosis was examined in the walls of the three largest yellow hierarchical follicles (F3-F1; F3