大绒鼠解偶联蛋白1基因部分序列扩增与分析

解偶联蛋白1（Uncoupling protein 1,UCP1）是位于褐色脂肪组织线粒体内膜上的一种解偶联蛋白,该蛋白可以诱导质子漏从而产热。通过设计简并引物进行RT-PCR从大绒鼠BAT中获得UCP1基因cDNA核心序列,RTPCR所得产物长约458 bp,包含的开放阅读框（open reading frame,ORF）为456 bp,编码151个氨基酸。通过BLAST搜索,所得大绒鼠UCP1基因cDNA氨基酸序列与黑线仓鼠、橙腹草原田鼠、金黄仓鼠、小家鼠和褐家鼠等哺乳动物的UCP1氨基酸序列同源性均在80%以上,而与鱼类和两栖类的氨基酸序列同源性在61%以下。研究结果表明UCP1在哺乳类中高度保守。同时,通过NJ方法以UCP1序列构建系统进化树表明大绒鼠与橙腹草原田鼠聚成一支,构成田鼠类分支。     

长短日照下绍鸭性成熟时间,下丘脑GnRH-Ⅰ、POMC和NPY mRNA表达的差异

利用长日照和短日照环境调控绍鸭的性成熟时间,并采用反转录多聚酶链式反应（RT．PCR）方法,检测两种日照组（n=6）鸭性成熟过程中下丘脑促性腺激素释放激素-Ⅰ（GnRH-Ⅰ）、阿片促黑激素皮质素原（POMC）和神经肽Y（NPY） mRNA表达的发育性变化及日龄间差异。结果表明,长日照组性成熟时间较短日照组提前30d。RT-PCR结果显示,两组鸭下丘脑GnRH-Ⅰ mRNA的发育性表达模式相似,从出雏至性成熟过程中,其表达水平呈逐渐增加趋势;长日照组下丘脑GnRH．ImRNA表达丰度均高于同日龄短日照组,并于120日龄时其差异达显著水平。两组POMC mRNA的表达丰度呈现先增加后降低趋势,于性成熟前后显著下降;日照长度对其基因表达具有显著影响。两组NPYmRNA表达水平分别于60和90日龄时显著升高,于性成熟时显著下降,但光照对其基因表达无显著影响。下丘脑GnRH-Ⅰ基因表达的显著上调可能是禽类性成熟启动的关键因素,也是长日照条件下绍鸭性早熟的主要原因;GnRH-Ⅰ表达的上调与下丘脑POMC和NPY mRNA表达变化有关。     

长短日照下绍鸭性成熟时间,下丘脑GnRH-I、POMC和NPY mRNA表达的差异

利用长日照和短日照环境调控绍鸭的性成熟时间,并采用反转录多聚酶链式反应(RT-PCR)方法, 检测两种日照组(n=6)鸭性成熟过程中下丘脑促性腺激素释放激素-Ⅰ(GnRH-Ⅰ)、阿片促黑激素皮质素 原(POMC)和神经肽Y(NPY)mRNA表达的发育性变化及日龄间差异。结果表明,长日照组性成熟时间较短 日照组提前30d。RT-PCR结果显示,两组鸭下丘脑GnRH-Ⅰ mRNA的发育性表达模式相似,从出雏至性成熟 过程中,其表达水平呈逐渐增加趋势;长日照组下丘脑GnRH-Ⅰ mRNA表达丰度均高于同日龄短日照组,并于 120日龄时其差异达显著水平。两组POMC mRNA的表达丰度呈现先增加后降低趋势,于性成熟前后显著下降; 日照长度对其基因表达具有显著影响。两组NPY mRNA表达水平分别于60和90日龄时显著升高,于性成熟时 显著下降,但光照对其基因表达无显著影响。下丘脑GnRH-Ⅰ基因表达的显著上调可能是禽类性成熟启动的关 键因素,也是长日照条件下绍鸭性早熟的主要原因;GnRH-Ⅰ表达的上调与下丘脑POMC和NPY mRNA表达变 化有关。     

NPY和POMC基因在生长差异与饥饿再摄食鳙个体中的表达分析

为研究摄食促进基因和摄食抑制基因对鱼类生长调控和饥饿再摄食过程的影响, 研究克隆了鳙神经肽Y(HynNPY)基因和前阿黑皮素原(HynPOMC)基因cDNA序列, 采用qRT-PCR技术分析它们在生长显著差异鳙个体下丘脑、肠中的基因表达变化; 设置对照组(连续投喂4周)、饥饿组、饥饿再摄食组, 分析NPY和POMC在不同处理组鳙的下丘脑、肠中的基因表达变化。鳙NPY和POMC基因cDNA全长分别为839和799 bp, 开放阅读框有291和657 bp, 分别编码96和218个氨基酸。系统进化分析结果表明, 鳙NPY和POCM基因具有高度保守性。鳙NPY在下丘脑的表达量最高, 其次为肠和脑; POMC在肠道中的表达量最高, 其次为下丘脑和肝脏。在相同环境下生长差异鳙个体的下丘脑和肠中, NPY在极大个体的表达量高于极小组个体, POMC在极小个体中的表达量高于极大个体。饥饿导致NPY在下丘脑表达上升, 在肠表达量显著上升, 恢复摄食后, NPY在下丘脑和肠中表达量下降; POMC在饥饿组下丘脑和肠中都表现为表达量呈显著上升, 复投喂后POMC表达量逐步下降至接近对照组水平。肠组织学观察显示, 极大个体的肠腔直径、肠绒毛长度、肠壁厚度均优于极小个体; 饥饿胁迫导致的肠绒毛断裂、黏膜白细胞浸润等损伤能在再摄食后逐步恢复。研究结果表明, NPY促进摄取更多食物, POMC抑制食欲, 二者共同参与鳙中枢神经和外周组织摄食调控, 并可能通过影响摄食行为、营养吸收和GH分泌从而直接或间接调控鳙生长。研究结果对加深鳙生长调控分子机制的理解及遗传改良具有较大的理论和现实意义。     

In vitro leptin treatment of rainbow trout hypothalamus and hindbrain affects glucosensing and gene expression of neuropeptides involved in food intake regulation

The aim of the present study was to evaluate in hypothalamus and hindbrain of rainbow trout in vitro the effect of leptin treatment on glucosensing capacity and the expression of orexigenic and anorexigenic peptides involved in the control of food intake. In a first experiment, the response of parameters involved in glucosensing (GK, PK and GSase activities; GK expression and glucose; glycogen and DHAP levels) and the expression of orexigenic (NPY) and anorexigenic (POMC, CART, CRF) peptides was assessed in hypothalami and hindbrain incubated for 1 h with 2, 4 or 8 mM d-glucose alone (controls) or with 10 nM leptin, or with 10 nM leptin plus inhibitors of leptin signaling pathways (50 nM wortmannin and 500 nM AG490). Leptin treatment increased levels in parameters involved in glucosensing. Leptin treatment decreased NPY mRNA levels in hypothalamus without affecting the expression of the other peptides assessed. Leptin effects were reverted in the presence of inhibitors for all parameters assessed suggesting the involvement of JAK/STAT and IRS-PI(3)K pathways. In a second experiment, we observed time-dependent (1-3 h) and dose (10, 20 and 50 nM)- effects of leptin treatment in decreasing NPY mRNA levels without affecting expression of the other peptides assessed. Considering the orexigenic action of NPY in fish, it seems that the anorexic effect of leptin can be mediated by reduced expression of NPY occurring in hypothalamus, and that change can be related to the activation of the glucosensing system occurring simultaneously.     

Central insulin sensitivity in male and female juvenile rats

The incidence of juvenile obesity is increasing at an alarming rate. In adults, central insulin administration decreases hypothalamic orexigenic neuropeptides, food intake and body weight more effectively in males than females. Mechanisms regulating energy balance in juvenile animals are inherently different from those in adults due to differences in growth rates and hormonal milieu. Therefore, we sought to determine if central insulin treatment in juvenile rats (4 wk) would have similar sex-dependent effects on food intake as those reported in adult rats. Twenty-four hour food intake was measured following icv saline or insulin (0.01 or 0.1 U) prior to the onset of dark phase of the light cycle. An additional set of animals was used to assess the effects of central insulin on hypothalamic orexigenic (NPY, AgRP) and anorexigenic (POMC) neuropeptide mRNA expression. In both males and females, insulin reduced meal size initially (first 4 h) and later decreased meal frequency (4-24 h) to reduce cumulative food intake. Consistent with this, central insulin decreased hypothalamic NPY and AgRP and increased POMC mRNA expression. In contrast to adult studies, there were no demonstrated sex differences. These studies indicate that juvenile females and males are equally sensitive to central insulin anorexigenic effects, perhaps due to a lack of circulating gonadal hormones. The anorexigenic responsiveness of both genders suggests a potential pharmacologic approach to childhood obesity.     

MCH and feeding behavior-interaction with peptidic network

Numerous works associate the MCH peptide, and the hypothalamic neurons that produce it, to the feeding behavior and energy homeostasis. It is commonly admitted that MCH is an orexigenic peptide, and MCH neurons could be under the control of arcuate NPY and POMC neurons. However, the literature data is not always concordant. In particular questions about the intrahypothalamic circuit involving other neuropeptides and about the mechanisms through which MCH could act are not yet clearly answered. For example, which receptors mediate a MCH response to NPY or alpha-MSH, does MCH act alone, is there any local anatomical organization within the tuberal LHA? A review of the current literature is then needed to help focus attention on these unresolved and often neglected issues.     

温度和增加胎仔数对大绒鼠哺乳期能量代谢的影响

为了阐明温度和增加胎仔数对大绒鼠Eothenomys miletus哺乳期能量代谢的影响,本研究测定了在不同温度条件下增加胎仔数（比正常胎仔数多）,大绒鼠的摄食量、胎仔数、胎仔质量、静止代谢率、非颤抖性产热和乳腺质量。结果表明：不同胎仔数对大绒鼠的摄食量、胎仔质量和产热能力没有影响。低温增加了母体的摄食量和产热能力,断奶时低温组的胎仔质量显著低于常温组。低温组和常温组大绒鼠的乳腺质量差异无统计学意义。研究结果说明低温可以增加大绒鼠摄食量,但是对乳腺质量没有影响,表明大绒鼠哺乳期的持续能量摄入上限可能受到乳腺分泌乳汁的限制,支持外周限制假说。     

大绒鼠消化道形态的季节变化

为探讨栖息于横断山地区大绒鼠(Eothenomys miletus)的消化道特征与环境之间的适应关系,对自然环境中大绒鼠消化道各项指标进行了测定.实验分别测定了不同季节大绒鼠胃、小肠、大肠、盲肠的长度,及其含内容物重、去内容物重、干组织重.结果表明,大绒鼠消化道特征存在季节性变化,随着温度降低、食物质量下降,大绒鼠的小肠、盲肠长度增加;各器官重量均在6月份最大;大绒鼠在低温、食物质量下降、繁殖等胁迫因子作用下,通过增加食物摄入、调节消化道形态来满足能量需求的增加,维持正常的生理机能.大绒鼠的消化道在不同季节中表现出的变化模式,与其低纬度高海拔、年平均温度较低的生存环境有关,从一方面反映了该物种在温度胁迫下的生存机制和适应对策.     

食物限制对雄性大绒鼠能量代谢特征的影响

为探讨横断山区大绒鼠适应食物匮乏的适应对策,将成年雄性大绒鼠随机分为自由取食组和饲喂正常摄食量的80% 限食组。测定了自由取食组和限食组雄性大绒鼠的体重、静止代谢率、非颤抖性产热以及体脂含量、血清瘦素含量、肝脏鲜重、褐色脂肪组织重量和消化道形态。结果显示:限食使雄性大绒鼠的体重、体脂含量、静止代谢率、非颤抖性产热、褐色脂肪组织重量和大肠、小肠长度显著降低,使盲肠内容物重量显著增加。血清瘦素含量与体重、体脂含量呈极显著正相关。在限食条件下,大绒鼠主要通过降低体重、基础代谢和产热的能量支出以及动用体内脂肪以应对食物资源短缺的环境条件,瘦素可能参与了能量代谢和体重的适应性调节。     

横断山脉大绒鼠最大代谢率的季节性差异

最大代谢率对于动物的生存、繁殖和分布具有关键作用。实验用开放式呼吸仪测定了大绒鼠(Eothenomys miletus)夏季和冬季的冷诱导最大代谢率和运动最大代谢率。大绒鼠夏季的冷诱导最大代谢率O2为(7.24±0.61)ml/(g.h),运动最大代谢率O2为(7.69±0.59)ml/(g.h);冬季的冷诱导最大代谢率O2为(8.61±0.42)ml/(g.h),运动最大代谢率O2为(7.51±0.51)ml/(g.h)。冷诱导最大代谢率冬夏之间具有差异显著,而运动最大代谢率则差异不显著。结果表明:栖息于横断山脉的大绒鼠由于受到低温的胁迫,导致其冷诱导最大代谢率季节性变化较大,而运动最大代谢率则相对稳定。     

Network of hypothalamic neurons that control appetite

The central nervous system (CNS) controls food intake and energy expenditure via tight coordinations between multiple neuronal populations. Specifically, two distinct neuronal populations exist in the arcuate nucleus of hypothalamus (ARH): the anorexigenic (appetite-suppressing) pro-opiomelanocortin (POMC) neurons and the orexigenic (appetite-increasing) neuropeptide Y (NPY)/agouti-related peptide (AgRP) neurons. The coordinated regulation of neuronal circuit involving these neurons is essential in properly maintaining energy balance, and any disturbance therein may result in hyperphagia/obesity or hypophagia/starvation. Thus, adequate knowledge of the POMC and NPY/AgRP neuron physiology is mandatory to understand the pathophysiology of obesity and related metabolic diseases. This review will discuss the history and recent updates on the POMC and NPY/AgRP neuronal circuits, as well as the general anorexigenic and orexigenic circuits in the CNS. [BMB Reports 2015; 48(4): 229-233]