齐口裂腹鱼PGC-1a基因编码区的克隆及其在肌肉组织中的表达

目的：获得齐口裂腹鱼过氧化物酶体增殖物激活受体Y辅助活化因子1a（PGC-1a）cDNA序列,探讨其在齐口裂腹鱼肌肉组织中的表达规律。方法：根据斑马鱼基因PGC-1a序列设计引物,提取齐口裂腹鱼肌肉组织总RNA,经RT-PCR扩增PGC-1a仪基因序列;利用半定量RT-PCR分析齐口裂腹鱼PGC-1a基因在红肌和白肌中的mRNA表达特性。结果：获得齐口裂腹鱼PGC-1a基因序列2633bp,GenBank登陆号为JNl95738,其中ORF为2631bp,编码876个氨基酸残基,与同鲤科的斑马鱼、草鱼和金鱼的同源性较高,为88％～93％,但与哺乳动物和禽类如人、小鼠、大鼠、猪、牛和鸡的同源性较低,为49％～50％。在基础状态下,PGC-1a在齐口裂腹鱼红肌中的表达显著高于白肌,禁食可显著诱导PGC-1a在红肌和白肌中的表达水平。结论-首次克隆得到齐口裂腹鱼PGC-1a基因序列,其在红肌中的表达显著高于白肌中,禁食可显著诱导其在红肌和白肌中的表达,为研究PGC-1a在齐口裂腹鱼肌肉中的作用提供了理论依据。     

野生和养殖裂腹鱼血液学指标的比较研究

对齐口裂腹鱼和重口裂腹鱼的野生和养殖个体血液学指标进行了比较研究.结果显示:重口裂腹鱼野生组的红细胞长径和短径、红细胞核长径和短径、嗜中性粒细胞长径和短径、淋巴细胞长径和短径、总蛋白和甘油三酯与齐口裂腹鱼野生组相应指标差异显著;齐口裂腹鱼野生组的嗜中性粒细胞长径、嗜中性粒细胞短径、淋巴细胞长径、淋巴细胞短径和淋巴细胞核长径显著大于养殖组;重口裂腹鱼野生组的红细胞短径明显小于养殖组重口裂腹鱼;齐口裂腹鱼野生组血液中TP、GLU和TG显著低于齐口裂腹鱼养殖组;重口裂腹鱼野生组TG、T-CHO与养殖组重口裂腹鱼差异显著.这些差异说明齐口裂腹鱼和重口裂腹鱼在生理适应方面存在差异,养殖活动对它们的生理状况有一定的影响.     

齐口裂腹鱼CDH5基因特性及其对温和气单胞菌感染的应答

探讨齐口裂腹鱼(Schizothorax prenanti)血管内皮黏附分子(Cadherin 5,CDH5)的基因特性。用生物信息学软件分析齐口裂腹鱼CDH5基因序列,用Real-time PCR检测CDH5基因在齐口裂腹鱼感染温和气单胞菌后0 h、24 h和48 h的表达变化。获得CDH5基因全长c DNA序列,Gen Bank登录号为KT329441。该序列长4 825 bp,开放阅读框长2 313 bp,编码770个氨基酸。蛋白预测结果显示,该蛋白相对分子量为85.19 k D,等电点为5.01。存在信号肽序列,二级结构以随机卷曲、延伸链、α-螺旋为主。齐口裂腹鱼CDH5氨基酸序列与斑马鱼同源性达74%,与人的相似性为43%。CDH5在感染温和气单胞菌后在各组织中均有表达。脾脏中CDH5基因在24 h表达量显著高于0 h和48 h(P0.05)。肝脏、肾脏和肌肉中CDH5在48 h的表达量均显著高于24 h(P0.05)。心脏和肠道中CDH5基因在48 h的表达量显著高于0 h(P0.05)。CDH5基因在可能参与了齐口裂腹鱼抗温和气单胞菌感染的免疫应答,为深入研究CDH5在齐口裂腹鱼中的功能奠定基础。     

饲料蛋白质和能量水平对齐口裂腹鱼生长、体组成和蛋白质利用的影响

齐口裂腹鱼（Schizothorax prenanti Tchang）隶属鲤科、裂腹鱼亚科,俗称“雅鱼”,为长江上游重要经济鱼类,也是川西高原特有的冷水性鱼类。其肉质细嫩,味道鲜美,是名贵野生经济鱼类。近年来由于过度捕捞、环境污染等因素的影响,齐口裂腹鱼种群数量不断下降,分布区域日趋缩小,而随着人们生活水平的不断提高,对齐口裂腹鱼等名优水产品的需求却在不断增加。为保护和持续利用这一野生动物资源,     

齐口裂腹鱼白介素-1β基因的克隆及表达谱

探讨齐口裂腹鱼白介素-1β(Interleukin-1β,IL-1β)基因的基本特点。用RT-PCR方法以齐口裂腹鱼脾脏c DNA作为模板扩增IL-1β基因,用生物信息学软件分析其序列,用RT-PCR方法检测IL-1β基因在齐口裂腹鱼6种组织的表达情况。克隆获得的IL-1β序列长1 252 bp,共编码276个氨基酸。Gen Bank序列号为KU886235。IL-1β蛋白相对分子量为31.25 k D,等电点5.45,预测为亲水性蛋白。二级结构主要结构元件为随机卷曲和延伸链,有4个N-糖基化位点和16个磷酸化位点。NJ法系统进化树显示,齐口裂腹鱼与鲤鱼、鲫鱼的亲缘关系最近。组织表达分析显示,IL-1β基因主要在脾脏和肾脏中表达。这为深入研究鱼IL-1β的生物学功能提供理论依据。     

猪H-FABP基因多态性与肌内脂肪含量的关联分析

以山西白猪为实验材料,采用索氏脂肪抽提法对其背最长肌的肌内脂肪含量进行了测定。根据心脏脂肪酸结合蛋白(heart fatty acid-binding protein,H-FABP)基因的PCR-RFLP多态性,应用SPSS软件,用最小二乘法拟合一般线性模型(GLM)分析了H-FABP基因的不同位点的基因型对肌内脂肪含量的影响。结果表明,基因型hh、dd、BB对肌内脂肪含量影响的效应值最大。证明H-FABP基因可以作为肌内脂肪含量的侯选基因。     

黄芪多糖对齐口裂腹鱼生长、体组成和免疫指标的影响

试验研究黄芪多糖对齐口裂腹鱼生长性能、体组成及免疫指标的影响。以450尾健康的齐口裂腹鱼[体重(6.98±0.43)g;体长(9.11±0.25)cm]为试验对象,随机分为5组(C1、C2、C3、C4、C5),每组3个重复,每重复30尾试验鱼。C1、C2、C3、C4、C5组分别投喂在等氮等能(蛋白质含量38.29%,能量15.73 mJ/kg)的基础料中分别添加0、0.02、0.04、0.06、0.08%的黄芪多糖制成5种试验饲料,养殖齐口裂腹鱼50d。结果表明:饲料中未添加黄芪多糖组的增重率(WGR)、特定生长率(SGR)、饲料蛋白效率(PER)均显著低于黄芪多糖添加组(P<0.05),而饵料系数(FCR)则显著高于黄芪多糖添加组(P<0.05)。当黄芪多糖添加水平为0.04%时,试验鱼的WGR、SGR、PER均达到最大(分别为110.31%、1.86%/d和182.07%),FCR最低(1.44),与其他各组差异显著(P<0.05);以WGR、SGR、PER、FCR为指标,利用直线和抛物线回归分析表明,齐口裂腹鱼生长性能最佳时黄芪多糖添加水平为0.045%—0.074%;对齐口裂腹鱼机体组成分析表明,黄芪多糖对鱼体粗灰分和水分影响不显著(P>0.05),黄芪多糖添加水平为0.06%时机体粗蛋白最高,但与黄芪多糖添加水平为0.04%时无明显差异(P>0.05);粗脂肪含量在黄芪多糖添加水平为0.04%时最高,与其他各组差异显著(P<0.05);未添加黄芪多糖组血清免疫酶活性显著低于黄芪多糖添加组,齐口裂腹鱼血清免疫酶活性在一定范围内随黄芪多糖的增加而增强。黄芪多糖添加水平为0.04%时,碱性磷酸酶(ALP)活性最高;酸性磷酸酶(ACP)活性在黄芪多糖添加水平0.06%时最大;黄芪多糖添加水平应在0.06%—0.08%时,溶菌酶(LSZ)、超氧化歧化酶(SOD活性趋于稳定。这说明黄芪多对齐口裂腹鱼的生长和免疫力有明显的促进作用。综合考虑齐口裂腹鱼生长性能和免疫能力最佳时黄芪多糖添加水平为0.04%—0.074%。     

绵羊肌肉H-FABP和PPARγ基因表达的发育性变化及其对肌内脂肪含量的影响

选取不同日龄的雄性哈萨克羊和新疆细毛羊,屠宰后取背最长肌,用索氏抽提法检测肌内脂肪（Intamuscular fat,IMF）含量,用荧光实时定量PCR法检测心脏脂肪酸结合蛋白（heart fatty acid-binding protein,H-FABP）和过氧化物酶体增殖物激活受体γ（peroxisome proliferator-activated receptorγ,PPARγ）基因表达的发育性变化极其对IMF含量的影响.结果表明：（1）随着日龄的增加,IMF含量在雄性哈萨克羊中持续上升,且在各生长期间的差异显著（P＜0.05）,而在新疆细毛羊的各生长期间无显著差异（P＞0.05）.雄性哈萨克羊的IMF含量在30～90日龄间极显著高于新疆细毛羊（P＜0.01）;（2）雄性哈萨克羊肌肉H-FABP基因的表达量在2日龄时最高,30日龄时降到最低,然后持续上升,各日龄间差异显著（P＜0.05）.新疆细毛羊中的表达量在2日龄时也最高,然后持续下降,到90日龄时降到了最低点,然后上升,各日龄间差异也显著（P＜0.05）;（3）雄性哈萨克羊肌肉PPARγ基因的表达量随着日龄的增加而呈下降趋势,各日龄间差异显著（P＜0.05）;新疆细毛羊中的表达量在2～60日龄期间持续上升且各日龄间差异显著（P＜0.05）,但在90日龄时降到最低点,然后上升;（4）在哈萨克羊中,H-FABP基因在30～90日龄间的表达量与IMF含量的相关系数为0.737（P＜0.01）,而PPARγ基因在2-90日龄间的表达量与之的相关系数为-0.835（P＜0.01）.     

基于mtDNA Cyt b序列分析齐口裂腹鱼群体遗传多样性

研究测定了长江上游4个齐口裂腹鱼(Schizothorax prenanti)野生群体(重庆巫溪、重庆城口、四川雅安、四川阿坝)共104个个体的线粒体Cyt b基因部分序列, 以探讨齐口裂腹鱼野生群体的遗传多样性和遗传结构。结果表明: 在104个个体Cyt b序列中共检测到43个多态性位点, 25个单倍型。4个齐口裂腹鱼群体的单倍型多样性介于0.704—0.884, 核苷酸多样性介于0.007—0.012。群体间Kimura双参数遗传距离介于0.008—0.017, 其中四川雅安群体与四川阿坝群体间遗传距离最近, 基因交流频繁。重庆城口群体与四川雅安群体间遗传距离最远, 基因交流受阻。AMOVA分析表明, 齐口裂腹鱼的遗传分化主要来自群体内部, 且组群间、组群内群体间和群体内存在显著的遗传分化。中性检验得到Tajima’s D和 Fu’s F_s的值不显著, 且歧点分布图呈多峰, 表明长江上游4个齐口裂腹鱼野生群体未经历过种群扩张。研究旨为齐口裂腹鱼野生资源保护提供必要参考意见, 同时为齐口裂腹鱼种质资源合理开发和利用提供理论依据。     

热应激对齐口裂腹鱼非特异性免疫功能及细胞凋亡的影响

为研究热应激及恢复对齐口裂腹鱼（Schizothorax prenanti）机体损伤、非特异性免疫功能和细胞凋亡的影响,测定了热应激前（19℃,对照组）、热应激（27℃,0、2h、4h、8h、12h,实验组）及恢复适温6h后（19℃,恢复组）的非特异性免疫相关生理生化指标及细胞凋亡率。结果表明,超氧化物歧化酶（Superoxide dismutase,SOD）在应激0、4h、8h、12h后活性上升,应激0和12h与应激前差异极显著（P<0.01）;髓过氧化物酶（Myeloper oxidase,MPO）在热应激4h达到最大值;谷草转氨酶（Glutamic-oxalacetic transaminase,GOT）的活性实验组与对照组差异不显著,恢复6h后GOT活性极显著高于对照组（P<0.01）;谷丙转氨酶（Glutamic-pyruvic transaminase,GPT）在应激0、4h、12h和恢复到适温6h后的活性极显著高于对照组（P<0.01）;丙二醛（Malondialdehyde,MDA）含量在热应激2h、4h和12h时极显著高于对照组（P<0.01）,8h时显著高于应激前（P<0.05）,应激12h MDA含量达到最大值,恢复6h后MDA含量恢复至应激前水平（P<0.05）;补体3（Complement 3,C3）含量升高,在热应激12h时显著高于对照组（P<0.05）,恢复6h C3含量下降,与对照组差异不显著（P>0.05）。持续27℃热应激不同时间后,各实验组的细胞凋亡率都显著高于对照组（P<0.05）,且在处理4h时细胞凋亡率达到最高值（52.42%）,恢复19℃ 6h后细胞凋亡率显著降低（P<0.05）。研究表明,持续27℃热应激改变了机体非特异性免疫力,导致鱼体出现炎症,损伤细胞,恢复到适温后,鱼体各项机能逐渐恢复。研究结果可为深入研究齐口裂腹鱼高温适应调控机理提供基础数据,也为齐口裂腹鱼养殖、运输等过程中的温度控制提供科学依据。     

齐口裂腹鱼线粒体DNA控制区结构分析

利用直接测序法对齐口裂腹鱼(Schizothorax prenanti)线粒体DNA控制区进行了测序,并对其序列结构进行了分析.结果表明,齐口裂腹鱼线粒体控制区碱基组成中碱基A和T的含A明显高于G和C的含量,所有类型碱基组成中碱基G的含量最低,这与其他硬骨鱼类控制区碱基组成一致.通过与哺乳类和鲤形目鱼类控制区序列进行对...     

Cloning and Tissue Expression of Chicken Heart Fatty Acid-Binding Protein and Intestine Fatty Acid-Binding Protein Genes

Fatty acid-binding proteins (FABPs) are members of a superfamily of lipid-binding proteins, occurring intracellularly in invertebrates and vertebrates. This study was designed to clone and characterize the genes of heart fatty acid-binding protein and intestine fatty acid-binding protein in the chicken. PCR primers were designed according to the chicken EST sequences to amplify cDNA of H-FABP and I-FABP genes from chicken heart and intestinal tissues. Analysis of sequence showed that the cDNA of the chicken H-FABP gene is 75 to 77% homologues to human, mouse, and pig H-FABP genes, and the chicken I-FABP gene is 71 to 72% homologues to human, mouse, and pig I-FABP genes. In addition, Northern blot analysis indicated that of the two genes, similar to the copartner of the mammal, H-FABP gene was expressed in a wide variety of tissues, and I-FABP gene was expressed only in intestinal tissues. The expression levels of the chicken H-FABP mRNA in heart and I-FABP mRNA in intestine had significant differences between the broilers from fat line and Bai'er layers at six weeks of age. The results of this study provided basic molecular information for studying the role of two FABPs in the regulation of fatty acid metabolism in avian species.     

Cloning and tissue expression of chicken heart fatty acid-binding protein and intestine fatty acid-binding protein genes

Fatty acid-binding proteins (FABPs) are members of a superfamily of lipid-binding proteins, occurring intracellularly in invertebrates and vertebrates. This study was designed to clone and characterize the genes of heart fatty acid-binding protein and intestine fatty acid-binding protein in the chicken. PCR primers were designed according to the chicken EST sequences to amplify cDNA of H-FABP and I-FABP genes from chicken heart and intestinal tissues. Analysis of sequence showed that the cDNA of the chicken H-FABP gene is 75 to 77% homologues to human, mouse, and pig H-FABP genes, and the chicken I-FABP gene is 71 to 72% homologues to human, mouse, and pig I-FABP genes. In addition, Northern blot analysis indicated that of the two genes, similar to the copartner of the mammal, H-FABP gene was expressed in a wide variety of tissues, and I-FABP gene was expressed only in intestinal tissues. The expression levels of the chicken H-FABP mRNA in heart and I-FABP mRNA in intestine had significant differences between the broilers from fat line and Bai'er layers at six weeks of age. The results of this study provided basic molecular information for studying the role of two FABPs in the regulation of fatty acid metabolism in avian species.     

齐口裂腹鱼幼鱼疲劳后游泳特性恢复状况研究

以中国地质公园（神农架）野生动物繁育基地提供的齐口裂腹鱼（Schizothorax prenanti）幼鱼（体长为14.017.5 cm,体重为39.665.6 g）为研究对象,利用自制鱼类游泳特性（Swimming performance）研究装置,运用递增流速法（Stepped velocity test）,研究了鱼类疲劳后20d和40d的游泳特性恢复状况。结果表明:（1）在20d内齐口裂腹鱼临界游泳速度（Critical swimming speed,Ucrit）相比于初次测试（20d前）有所降低,20d内的恢复状况较差;而在40d后,临界游泳速度已经基本恢复到最初状况,40d内的恢复状况较好。（2）疲劳后鱼类游泳效率显著下降,并且在40d后无法恢复正常。虽然40d后速度指数低于20d后速度指数,40d后游泳效率高于20d后游泳效率,但是相对于初次测试,40d后游泳效率恢复效果并不明显。（3）齐口裂腹鱼在疲劳后40d内,恢复能力（Recovery capability）无法恢复到正常状况,但由于3次测试中疲劳后过量耗氧（Excess post-exercise oxygen cost,EPOC） 波动并不大,说明疲劳对其恢复能力影响并不大。（4）20d后和40d后无氧呼吸时间均较第一次测试时提前,说明疲劳损伤对其有氧呼吸能力产生了持续影响。（5）疲劳对鱼类生态行为（Ecological behavior）影响不明显。研究有利于为鱼道建设提供基础资料,对生物多样性保护具有深远意义。       

齐口裂腹鱼味蕾及上皮细胞的扫描电镜观察

本文报道扫描电镜对齐口裂腹鱼的味蕾在须、口咽腔、食道的分布和表面形态的观察。结果表明在须、唇、颌、口腔顶壁、咽、舌、食道均有味蕾分布,且在表面形态上呈现多态性。上皮细胞表面有微脊。     

Characterization of Schizothorax prenanti cgnrhII gene: fasting affects cgnrhII expression

In this study, the role of chicken gonadotropin‐releasing hormone II (cgnrhII) in feeding regulation was investigated in Schizothorax prenanti. First, the full‐length S. prenanti cgnrhII cDNA consisted of 693 bp with an open reading frame of 261 bp encoding a protein of 86 amino acids. Next, cgnrhII was widely expressed in the central and peripheral tissues. Last, there were significant changes in cgnrhII mRNA expression in the fasted group compared to the fed group in the S. prenanti hypothalamus during 24 h fasting (P < 0·05). Furthermore, the cgnrhII gene expression presented a significant decrease in the fasted group compared with the fed group (P < 0·05) on days 3, 5 and 7, after re‐feeding, there was no significant changes in cgnrhII mRNA expression level between refed and fed group on day 9 (P > 0·05). Thus, the results suggest that cGnRH II expression is influenced by fasting and the gene may be involved in feeding regulation in S. prenanti.     

齐口裂腹鱼肠粘膜上皮透射电镜观察

作用用透射电镜观察了齐口裂腹鱼肠粘膜上的皮的超微结构,肠上皮共有五种细胞,吸收细胞,杯状细胞,内分泌细胞,巨噬细胞,淋巴细胞等,文中详细描述了这些细胞的形态结构,并对其结构与功能之间的关系进行了初步讨论。     

Polymorphism of heart fatty acid-binding protein gene associatied with fatness traits in the chicken

Fatty acid-binding proteins (FABP) belong to a superfamily of lipid binding proteins that exhibit a high affinity for long chain fatty acids and appear to function in metabolism and intracellular transportation of lipids. The current study was designed to investigate the effects of heart (H)-FABP gene on chicken growth and body composition traits. The Northeast Agricultural University divergent broiler lines for abdominal fat and a broiler X silkie F2 population were used in this study. Body weight and body composition traits were measured in the populations. Primers were designed according to the chicken H-FABP gene sequence. Polymorphisms between parental lines were detected by DNA sequencing. PCR-RFLP and PCR-fragment length polymorphism methods were developed to genotype the populations. The results showed that the H-FABP gene polymorphisms in the two populations were associated with abdominal fat percentage. It implied that H-FABP gene can be a candidate locus or linked to a major gene(s) that affects abdominal fat content in the chicken.