Effects of "Bioactive" amino acids leucine, glutamate, arginine and tryptophan on feed intake and mRNA expression of relative neuropeptides in broiler chicks

ABSTRACT: Feed intake control is vital to ensuring optimal nutrition and achieving full potential for growth and development in poultry. The aim of the present study was to investigate the effects of L-leucine, L-glutamate, L-tryptophan and L-arginine on feed intake and the mRNA expression levels of hypothalamic Neuropeptide involved in feed intake regulation in broiler chicks. Leucine, glutamate, tryptophan or arginine was intra-cerebroventricularly (ICV) administrated to 4d-old broiler chicks respectively and the feed intake were recorded at various time points. Quantitative PCR was performed to determine the hypothalamic mRNA expression levels of Neuropeptide Y (NPY), agouti related protein (AgRP), pro-opiomelanocortin (POMC), melanocortin receptor 4 (MC4R) and corticotrophin releasing factor (CRF). Our results showed that ICV administration of L-leucine (0.15 or 1.5 mumol) significantly (P < 0.05) increased feed intake up to 2 h post-administration period and elevated both hypothalamic NPY and AgRP mRNA expression levels. In contrast, ICV administration of L-glutamate (1.6 mumol) significantly (P < 0.05) decreased feed intake 0.25, 0.5 and 2 h post-injection, and increased hypothalamic CRF and MC4R mRNA expression levels. Meanwhile, both L-tryptophan (10 or 100 mug) and L-arginine (20 or 200 mug) had no significant effect on feed intake. These findings suggested that L-leucine and L-glutamate could act within the hypothalamus to influence food intake, and that both orexigenic and anorexigenic Neuropeptide genes might contribute directly to these effects.     

Melanocortin 2 receptor antagonists in canine pituitary-dependent hypercortisolism: in vitro studies

Canine hypercortisolism is most often caused by an ACTH-secreting pituitary adenoma (pituitary-dependent hypercortisolism; PDH). An interesting target for a selective medical treatment of PDH would be the receptor for ACTH: the melanocortin 2 receptor (MC2R). In this study we investigated whether two peptide compounds, BIM-22776 (#776) and BIM-22A299 (#299), are effective MC2R antagonists in vitro. Their effects on cortisol production and mRNA expression of steroidogenic enzymes, MC2R and melanocortin 2 receptor accessory protein (MRAP) were evaluated in primary adrenocortical cell cultures (n?=?8) of normal canine adrenal glands. Cortisol production stimulated by 50 nM ACTH was dose-dependently inhibited by #299 (inhibition 90.7?±?2.3% at 5 μM) and by #776 (inhibition 38.0?±?5.2% at 5 μM). The ACTH-stimulated mRNA expression of steroidogenic enzymes, MC2R and MRAP was significantly inhibited by both compounds, but most potently by #299. These results indicate that canine primary cell culture is a valuable in vitro system to test MC2R antagonists, and that these compounds, but especially #299, are effective MC2R antagonists in vitro. To determine its efficacy in vivo, further studies are warranted. Antagonism of the MC2R is a promising potential treatment approach in canine PDH.     

The immediate effects of weaning stress on the hypothalamus‐pituitary‐adrenal alteration of newly weaned piglets

This study was conducted to examine the effects of weaning stress on gene expression of specific markers in hypothalamus‐pituitary‐adrenal (HPA) axis and neuronal activity in the newly weaned piglets. Twelve 28‐days‐old, newly weaned crossbred (Landrace × Yorkshire × Duroc) male piglets from 6 l (2 piglets/l) were randomly categorized into two groups: (a) weaning stress: piglets were separated from their dams, relocated and mixed with the unacquainted domestic piglets for 2 hr (stress, n = 6); (b) no‐stress: piglets stayed with their dams in the farrowing house (NS; n = 6). After weaning stress, all piglets were electrically euthanized and the blood samples/HPA tissues were collected for subsequent analysis, including plasma cortisol and mRNA expression of c‐fos, c‐jun, corticotropin‐releasing hormone (CRH), CRH receptor 1 (CRHR‐1) and adrenocorticotropin hormone receptor (MC2R). Results: Weaning stress significantly (p < 0.05) increased the plasma cortisol level and suppressed the expression of c‐fos and CRH in hypothalamus. In addition, weaning stress enhanced the mRNA abundance of c‐jun and CRHR‐1 in the pituitary gland. No significant differences in the gene expression of MC2R and CRHR‐1 were observed in the adrenal gland between treatment groups. Taken together, HPA involved in weaning stress and CRHR‐1 and c‐jun could be potential markers to evaluate the activation of HPA axis.     

Brain interleukin-1 is involved in blood interleukin-6 response to immobilization stress in rats

Interleukin (IL)-6, a cytokine for host defense responses to infection and inflammation, is known to be induced by non-invasive physical or psychological stress, too. To test possible involvement of brain IL-1 in the stress-induced IL-6 production, IL-1 mRNA expression in the hypothalamus, in parallel with blood IL-6 level, was examined in rats subjected to restriction of their movement (immobilization stress). When rats were immobilized, the hypothalamic IL-1 beta mRNA level was increased in 1 hr, followed by progressive rises in the serum IL-6 level. The immobilization-induced rise in serum IL-6 was mimicked by intracerebroventricular (icv) administration of IL-1 beta under normal conditions, whereas it was attenuated by icv injection of an IL-1 receptor antagonist. These results indicate that IL-1 in the hypothalamus plays a pivotal mediating role in the stress-induced peripheral IL-6 production.     

Functional and phylogenetic analyses of a melanocortin-4 receptor mutation in domestic pigs

The melanocortin-4 receptor (MC4R), a G protein-coupled seven-transmembrane receptor, which is expressed in the brain, plays an important role in the control of mammalian energy homeostasis. A missense mutation (Asp298Asn) was identified in the porcine MC4R gene, which is associated with growth and food intake traits. The Asn298 mutation occurs within a highly conserved motif, NPLIY, of all members of G protein-coupled receptors; whereas, Asp298 is conserved in all five melanocortin receptor subtypes. Functional analysis of the porcine MC4R variant was performed with an in vitro gene expression system in 293 cells. Ligand binding (NDP-alphaMSH) did not differ between Asp298 and Asn298 MC4R proteins. However, the Asn298 MC4R variant was unable to stimulate cAMP production in response to NDP-alphaMSH stimulation; whereas, the Asp298 variant could stimulate cAMP accumulation. These results demonstrate that the Asp298 is required for normal MC4R signaling to the adenylyl cyclase. Sequencing of the MC4R gene of seven diverse genera within the Suiformes that include Hippopotamidae (hippos), Tayassuidae (peccaries) and Suidae (pigs), revealed 62 nucleotide variations in MC4R. Phylogenetic relationships of MC4R variations are consistent with those previously described from morphological and physiological data among the subfamilies of the Suiformes. These findings revealed that a single missense mutation (Asp298Asn) of aspartic acid (Asp) to asparagine (Asn) in MC4R gene decreased cAMP content and MC4R signaling, but with no difference in the ligand binding was associated with growth and feed intake traits in domestic pigs.     

Central insulin suppresses feeding behavior via melanocortins in chicks

Growing evidence suggests that insulin interacts with both orexigenic and anorexigenic peptides in the brain for the control of feeding behavior in mammals. However, the action of central insulin in chicks has not yet been identified. In the present study, we investigated the effects of central injection of insulin on feeding behavior in chicks. Intracerebroventricular (ICV) administration of insulin, at doses that do not influence peripheral glucose levels, significantly inhibited food intake in chicks. Central injection of insulin in chicks significantly increased expression of pro-opiomelanocortin (POMC) mRNA, and decreased that of neuropeptide Y (NPY) mRNA. Finally, co-injection of the melanocortin antagonist (SHU9119 or HS014) prevented the reduction in food intake caused by ICV administration of insulin. These data suggest that insulin functions in chicks as an appetite-suppressive peptide in the central nervous system, and that the central melanocortin system mediates this anorexic effect of insulin, as in mammals.     

Expression of skin color genes in lymphocytes of Karan Fries cattle and seasonal relationship with tyrosinase and cortisol

To evaluate difference in the expression of skin color genes (melanocortin 1 receptor (MC1R) and premelanosome (PMEL)) in lymphocytes during winter and summer season and their correlation with tyrosinase enzyme and cortisol, ten Karan-Fries heifers were selected from National Dairy Research Institute (NDRI) cattle farm. Blood samples were collected from the animals during winter (THI?=?60) and summer (THI?=?83) season at weekly intervals. Relative MC1R and PMEL messenger RNA (mRNA) expression of Karan Fries cattle was found to be significantly (P?P?P?相似文献   

MC4R基因研究进展

黑素皮质素受体-4(melanocortin-4 receptor,MC4R)是下丘脑腹内侧核分泌的一类肽类物质,为黑素皮质素受体家族5个亚型(MC1-5R)之一。在哺乳动物中,MC4R具有介导瘦蛋白(leptin)的功能,是一个调节能量平衡与能量动态平衡的重要信号分子,可与其内源性配体黑素皮质素激素(melanocortin,MC)或刺鼠色蛋白(agouti protein,agouti蛋白)和agouti相关蛋白(agouti related protein,AGRP)相结合,从而在控制食欲和体重稳态中起关键作用。因此,MC4R在人类肥胖研究中作为重要的调节因子倍受关注。最近有研究表明:MC4R的1232位点的G→A的突变与绵羊背膘厚度间存在关联,AG和AA型较GG型具有较高的背膘厚度。     

The A y allele at the agouti locus enhances sensitivity to endotoxin-induced lethality in mice

In the course of investigations on anorexia during infection, I found that B6-A(y) mice had significantly increased sensitivity to lipopolysaccharide (LPS)-induced lethality as compared with isogenic B6 mice. I also found that the sensitivity to the lethal effect of LPS dramatically increased in aged mice (age effect), both B6 and B6-A(y). However, the A(y) effect of enhancing sensitivity to LPS-induced lethality was still significant, suggesting that the A(y) effect is independent of age. In the absence of tumor necrosis factor alpha (TNFalpha), the A(y) effect was still significant, suggesting that the A(y) effect is independent of TNFalpha toxicity. A dose of LPS of 100 mug per mouse caused 15% lethality in B6, 65% in B6-A(y) (significantly higher than B6), and 100 % in leptin-deficient B6-ob/ob (significantly higher than B6 and B6-A(y)). The results support the hypothesis that endogenous leptin has a protective role against infection, and that a part of this leptin effect is mediated by alpha-melanocyte-stimulating hormone (alphaMSH). In contrast to the results of simple blockade at the melanocortin 4 receptor (MC4R), B6-A(y) suffered more severe LPS-induced anorexia than did B6; therefore, the pathway involving MC4R is not absolutely required for the LPS-induced anorexia, and the presence of pathways involving other melanocortin receptor types was suggested. Because alphaMSH is suggested to be an endogenous anti-inflammatory peptide, and because melanocortin 1 receptor (MC1R) is expressed in various cutaneous cell types, the A(y) effect might be caused via the pathway involving MC1R. Physiologic significance of alphaMSH-MC1R interaction in host defense against infection is discussed.     

鹅黑素皮质素受体-4基因的克隆与序列分析

黑素皮质素受体-4（Melanocortin receptor-4,MC4R）是其黑素皮质素受体MCR（MC1-5R）家族成员之一,属G蛋白偶联受体。它可与瘦蛋白、神经肽、α-黑素细胞刺激素等一起调节动物体重和采食量。参考鸡MC4R基因序列设计引物,克隆并测序了鹅MC4R基因。结果表明,鹅MC4R基因编码区全长996 bp,其核苷酸序列与鸡的同源性为95.3%,与人、牛、猪等哺乳动物同源性在75%-79%;其氨基酸序列与鸡的同源性达到98.5%。构建哺乳类、鸟类和鱼类MC4R基因核苷酸进化树显示,鹅较早地与哺乳类动物分化开来。分析MC4R蛋白的氨基酸残基特性参数表明,MC4R的7次跨膜结构与MC4R的亲水性区域、电荷密度以及氨基酸残基位于表面概率的变化规律相一致。     

Effects of age and strain on the expression of leptin receptor, neuropeptide Y and pro-opiomelanocortin in the hypothalamus of young chickens

1. An experiment was conducted to study changes in the expression of the hypothalamic leptin receptor, neuropeptide Y (NPY) and proopiomelanocortin (POMC) with age during the early neonatal period in two different strains of chickens: Beijing-You (BY) and Arbor Acres (AA). 2. Compared with BY chickens, AA chickens ate more, and grew faster. Hypothalamic NPY concentrations of both strains increased with age until d 7 followed by a decline. Hypothalamic NPY of BY chickens on d 7 was lower than in AA chickens at the same age. 3. No difference with age was observed in hypothalamic α-melanocyte-stimulating hormone (α-MSH) of BY chickens, while hypothalamic α-MSH in AA chickens on d 0 was higher than on the other days. Compared with AA chickens, BY chickens showed lower hypothalamic α-MSH on d 0. 4. Similar developmental changes between two strains were observed in the expression of leptin receptor, NPY or POMC genes in the hypothalamus. Hypothalamic mRNA of leptin receptor on d 0 was higher than on d 1 and 7. Unlike NPY, hypothalamic NPY mRNA on d 0 was higher than on the other days. 5. Hypothalamic POMC mRNA decreased gradually with age until 7 d followed by a slight increase. 6. The results showed that the developmental changes of hypothalamic signal molecules varied with age and strain. NPY, α-MSH and leptin receptor might be involved in the early programming of feed intake in newly hatched chickens.     

Study on Melanocortin-4 Receptor mRNA Expression in Goats Grazed in Leigongshan Mountain Area

In order to understand the differential expression among tissues or breeds of grazed goat,β-actin gene was used as control,the expression levels of melanocortin-4 receptor (MC4R) mRNA in 7 tissues of Qiandongnan small Xiang goat,Guizhou White goat,Guizhou Black goat,Qianbei Ma goat and Nanjiang Yellow goat were detected by Real-time fluorescence quantitative PCR technique with double standard curve method.The results showed that MC4R mRNA was expressed in all 7 tissues of 5 grazed goat breeds,the expression levels of subcutaneous fat and kidney were the highest,the expression levels of liver was the second,the expression levels of lung and heart were the third,and the expression levels of semimembranosus muscle and longissimus muscle were the lowest.There were no differences of the expression level of MC4R mRNA in liver among different breeds.In subcutaneous fat,kidney,lung,heart,semimembranosus muscle and longissimus muscle,the highest expression levels of MC4R mRNA were all observed in Guizhou White goat.The results suggested that for grazed goats,the expression levels of MC4R mRNA existed tissue differences,and except for the liver,it existed breed differences,too.This research laid a basis for molecular marker assisted selection of growth traits,carcass traits,and meat quality traits of grazed goat.     

Involvement of activin and inhibin in the regulation of food and water intake in the rat

The expression of activin and inhibin has been demonstrated in the hypothalamus, but their physiological roles in the brain remain to be elucidated. In the present study, involvement of activin and inhibin in the regulation of food and water intake was examined. Male rats were deprived of food or water for 12 and 60 hr, and mRNA levels of activin/inhibin alpha, betaA and betaB subunits in the hypothalamus were estimated by RT-PCR. Gene expression of alpha subunit transiently decreased at 12 hr of food deprivation, while it did not change during water deprivation. Food and water deprivation for 60 hr increased mRNA levels of betaA and betaB subunits, respectively. These results indicated that gene expression of each subunit was independently regulated. Injection of activin A (0.5 and 4.0 microg) into the third ventricle decreased food intake. Water intake was suppressed by 4.0 microg, but not 0.5 microg, of activin A. Intracerebroventricular injection of inhibin A (0.5 and 4.0 microg) decreased water intake in a dose dependent manner without affecting food intake, suggesting that inhibin could act independently of activin. Taken together, it is suggested that activin and inhibin take part in the central regulation of nutrient and fluid balance, though further study is needed to determine precise molecular species involved.     

冷应激下民猪黑素皮质素受体基因的表达变化

黑素皮质素受体3（melanocortins 3 receptors,MC3R）是G-蛋白耦联受体超家族的成员之一。研究采用Real-time PCR的方法,对冷应激下民猪腿肌内MC3R基因的表达变化情况进行了研究。结果表明,冷应激下,MC3R基因的表达出现了显著地上升（P〈0.05）。据此推测,民猪很可能是通过上调MC3R基因的表达来增加能量消耗而抵御寒冷气候的。     

新民猪RYR1和MC4R基因的多态性检测分析

为了调查兰尼定1型受体(RYR1)基因和黑素皮质素受体4(MC4R)基因[其目前公认的数量性状基因座(QTN)]在新组建的民猪群体内的分布情况,试验采用PCR-RFLP的方法,对黑龙江省农业科学院畜牧研究所收集整理的23头民猪的RYR1基因和MC4R基因进行检测,并与其他已报道的猪种进行比较。结果表明:RYR1基因的N等位基因频率为86.96%,n等位基因频率为13.04%;MC4R的A等位基因频率为76.19%,G等位基因频率为23.81%,与其他猪种相比,n等位基因频率偏高。     

Correlation analysis of hypothalamic mRNA levels of appetite regulatory neuropeptides and several metabolic parameters in 28‐day‐old layer chickens

Various lines of evidence suggest that appetite‐related neuropeptides in the hypothalamus are regulated by adiposity signals such as leptin and insulin in mammals. In the present study, we examined age‐dependent changes in the weight of abdominal fat and hypothalamic mRNA levels of neuropeptide Y (NPY, an orexigenic neuropeptide) and proopiomelanocortin (POMC, a precursor of anorexigenic neuropeptides) in growing chickens at 7, 14, 21 and 28 days of age. Hypothalamic NPY mRNA levels were significantly (P < 0.05) decreased after 14 days of age, whereas hypothalamic POMC mRNA levels were significantly (P < 0.05) increased at 28 days of age. The percentage of abdominal fat was significantly increased after 14 days of age in chickens. We next examined the correlation of hypothalamic NPY and POMC mRNA levels and several parameters at 28 days of age. There were no significant correlations between hypothalamic mRNA levels of NPY or POMC and the percentage of abdominal fat. These findings suggest that the gene expressions of NPY and POMC do not depend on adiposity in chickens, at least in 28‐day‐old layer chickens.     

RFRP-3对大鼠采食行为和生长性能的影响

为了探讨RFRP-3对大鼠采食行为以及生长性能的影响,选取体重相近的清洁级SD大鼠30只,随机分为3组,分别为对照组[腹腔注射生理盐水200 μL/(次·只)]、RFRP-3低剂量组[腹腔注射1 μg/100 μL RFRP-3,200 μL/(次·只)]和高剂量组[腹腔注射10 μg/100 μL RFRP-3,200 μL/(次·只)],每组10只,雌、雄各半,每天注射2次,连续腹腔注射14 d。观察和记录大鼠的采食量、采食行为和体重变化,计算料肉比及Lee指数,分离血清并检测血脂相关指标,并对下丘脑食欲相关因子的表达水平进行检测。结果表明,与对照组相比,仅在光照环境中腹腔注射高剂量RFRP-3会导致大鼠采食量极显著(P<0.01)增加;采食行为方面主要表现为采食频率极显著(P<0.01)增高和饱腹率明显下降;生长性能方面主要表现为体重增长极显著(P<0.01)加快以及料肉比显著(P<0.05)降低;在肥胖指标中,Lee指数显著(P<0.05)增加,血清中甘油三酯和总胆固醇的含量极显著(P<0.01)升高;下丘脑NPY mRNA表达水平显著(P<0.05)上升,而POMC mRNA表达水平显著(P<0.05)下降。试验表明,腹腔注射RFRP-3可促进大鼠的采食行为并提高其生长性能,且这种生理功能的发挥可能是通过调节下丘脑食欲相关因子的变化完成的。     

L-精氨酸对大鼠胃肠激素分泌及食欲的影响

本试验旨在研究L-精氨酸(L-Arg)对大鼠胃肠道钙敏感受体(calcium-sensing receptor,CaSR)基因表达、胃肠激素分泌及采食量的影响,试验分为两个部分。试验I,选择32只SD雄性大鼠,随机分为4组,即对照组(灌胃0 mmol·kg^-1 L-Arg·HCl),低剂量组(5 mmol·kg^-1 L-Arg·HCl),中剂量组(10 mmol·kg^-1 L-Arg·HCl)和高剂量组(20 mmol·kg^-1 L-Arg·HCl),记录大鼠灌胃后不同时间点采食量,筛选显著影响大鼠采食的L-Arg最适剂量用于试验II。试验II,选择30只雄性大鼠随机分为两组,L-Arg·HCl高剂量组(20 mmol·kg^-1)及对照组(0 mmol·kg^-1),灌胃大鼠一周(每天一次),记录大鼠体重变化及日采食量。试验结束后采集血液、胃、小肠及下丘脑组织,检测血液中胃肠激素水平,胃肠道CaSR及下丘脑食欲调节因子基因表达。结果显示:试验I,10和20 mmol·kg^-1的L-Arg·HCl分别在灌胃后的0~1 h和0~24 h显著降低大鼠采食量(P<0.05),其中20 mmol·kg^-1效果最显著;试验II,与对照组相比,20 mmol·kg^-1 L-Arg·HCl显著降低了大鼠体增重及前两天的累加采食量(P<0.05),促进了胃肠道CaSR、厌食因子POMC基因表达和胆囊收缩素(CCK)分泌(P<0.05),胃泌素(gastrin)及葡萄糖依赖性促胰岛素多肽(GIP)的分泌虽有增加但差异不显著。相关分析结果显示,血清中CCK浓度与十二指肠、空肠CaSR及下丘脑POMC基因表达量呈显著正相关(P<0.05);GIP浓度与十二指肠CaSR及下丘脑POMC基因表达量呈显著正相关(P<0.05);GLP-1浓度与空肠CaSR及下丘脑POMC基因表达量有正相关的趋势(0.05相似文献