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1.
为了解鲤白细胞介素17B基因(IL-17B)的功能,实验使用同源搜索和基因克隆技术在鲤基因组中挖掘到2个IL-17Bs基因(CcIL-17B1和CcIL-17B2),均有3个外显子和2个内含子,编码198个氨基酸,内含IL-17家族特有的由4个半胱氨酸形成的2个二硫键,蛋白序列一致性高达91.92%。共线性分析显示,在硬骨鱼类染色体加倍过程中,IL-17B及其附近基因出现了丢失,大部分硬骨鱼类有1个IL-17B、斑马鱼2个IL-17Bs均丢失,而鲤特有的染色体加倍致使存在2个CcIL-17Bs。实时荧光定量PCR (qPCR)结果显示,CcIL-17Bs在鲤受精卵发育早期(0~12 h)和成鱼性腺中高表达。使用大肠杆菌表达系统,获得了可溶的重组蛋白NusA-17B。肛灌不同浓度的NusA-17B,结果显示,高浓度(500μg/kg)组的1和3 d的鲤肠道组织肠绒毛缺损,出现大量的杯状细胞和炎性细胞,定量分析显示,炎症因子基因IL-1β、IFN-γ、IL-6、趋化因子CCL20和NF-κB的表达均显著上调;7 d时肠道组织结构和炎症相关基因的表达均得到恢复,与对照组无显著差异。低浓度(5...  相似文献   

2.
青海湖裸鲤是唯一一种适应青海湖高盐碱水环境的鲤科鱼类。苯丙氨酸-X-酪氨酸-天冬氨酸(FXYD)家族是一类小分子单跨膜蛋白,具有调节离子通道作用,在多种生物的盐度胁迫和盐度适应中发挥重要作用。笔者基于青海湖裸鲤全基因组数据,利用生物信息学方法,鉴定到16个青海湖裸鲤FXYD(gpFXYD)基因,通过多重序列比对和系统进化分析,16个gpFXYD基因成员分属于5个亚型。共线性分析表明,青海湖裸鲤FXYD基因家族中12个成员起源于全基因组复制,且与斑马鱼FXYD基因存在共线性,推测全基因组复制导致了青海湖裸鲤FXYD基因的加倍。分子进化分析显示,gpFXYD1b/1c和gpFXYD5c/5d基因的进化选择压力(Ka/Ks)>1,表明其在进化过程中受到了正选择,可能发生了适应性进化。不同亚家族在青海湖裸鲤多种组织表达中,gpFXYD1s基因在青海湖裸鲤的心脏和脑组织特异性表达,而gpFXYD7s基因在青海湖裸鲤的脑组织中特异表达,分析gpFXYD基因不同亚型可能在不同组织中发挥功能。组织表达中,gpFXYD5s、gpFXYD6s、gpFXYD11s基因在青海湖裸鲤肾脏和鳃组织中出现显著...  相似文献   

3.
孙婷  刘伟  徐鹏  孙效文 《水产学报》2012,36(2):180-190
Wnt1诱导分泌蛋白(WISP)基因家族与CYR61、CTGF、NOV基因共同构成了CCN家族。研究通过鲤基因组序列与斑马鱼WISP基因编码区全序列的比对获得WISP1a、WISP1b、WISP2、和WISP3等4条序列。经过克隆、测序、比对拼接得到其开放阅读框,分别是1 089、1 077、1 038和1 026 bp,它们都是由5个外显子和4个内含子构成,分别编码362、358、345和341个氨基酸。分子系统学分析表明,鲤4个WISP基因具有高度同源性,其中WISP1a、WISP1b和WISP2均含有4个模块,WISP3缺少第2个模块。通过RT-PCR检测WISP基因在鲤组织中的表达,结果表明,WISP1a鲤在13个组织中均有表达,皮肤中表达最高,脾、肠、卵巢次之,其他组织中表达较低。WISP1b在精巢、脑、皮肤、卵巢中表达较高。WISP2在血液、鳃中表达较高。WISP3在血液、脑、肝中表达较高。实时荧光定量PCR法分析WISP基因在鲤胚胎发育时期的表达,结果表明,除WISP3外,WISP1a、WISP1b和WISP2在前期表达较高,36 h表达最低,随后逐渐升高至第6天开始下降。  相似文献   

4.
为探究刺鼠相关蛋白/神经肽Y (AgRP)神经元在金钱鱼应答饥饿与复投喂过程中所起的调控作用,采用反转录PCR自金钱鱼下丘脑克隆了agrp1和agrp2 2种亚型基因,并采用实时荧光定量PCR分析了2种基因在金钱鱼不同组织中的表达情况和正常投喂组(2、7 d)、饥饿组(2、7 d)、复投喂组(饥饿7 d然后复投喂3 h)金钱鱼(体质量52~60 g)下丘脑、肝脏和肠道中agrp1和agrp2基因的表达情况。序列分析结果显示,agrp1和agrp2基因的开放阅读框长度分别为429 bp和351 bp。组织表达结果显示,agrp1基因主要在下丘脑和肌肉中表达,agrp2基因主要在下丘脑和脑垂体中表达。饥饿及复投喂结果表明:下丘脑中,agrp1基因表达水平在饥饿2 d后差异不显著,饥饿7 d后显著下调,复投喂后其表达水平恢复正常;肝脏中,agrp1基因表达水平在饥饿2 d后下调,但饥饿7 d后显著上调,复投喂后其表达水平下调到正常水平;肠道中,agrp1基因表达水平在饥饿2 d后无显著差异,饥饿7 d后显著上调,复投喂后其表达水平下调到正常水平。饥饿和复投喂期间,下丘脑、肝脏和肠道中agrp...  相似文献   

5.
通过同源克隆和染色体步移对光倒刺鲃(Spinibarbus hollandi)黑素皮质素受体-5 (melanocortin receptor-5,mc5r)基因进行了克隆,应用生物信息学方法对mc5r基因序列和推测的氨基酸序列进行分析,采用荧光定量PCR技术研究其m RNA的组织分布、日周期变化以及饥饿再投喂的表达情况。结果表明mc5r基因长度为1 947 bp,含有1个987 bp的外显子,编码329个氨基酸。通过序列同源比对及系统发育进化分析,光倒刺鲃mc5r的氨基酸序列与鲤(Cyprinus carpio)、犀角金线鲅(Sinocyclocheilus rhinocerous)的mc5r同源性达到98%。光倒刺鲃mc5r在大脑中显著表达,在间脑、心脏的表达量次之,在中脑、小脑表达量较少,在肾脏、胃、肌肉、脾、肠、性腺、肝脏、延脑、眼中微量表达。光倒刺鲃mc5r mRNA的日周期表达量随昼夜节律而变化,饥饿再投喂实验发现饥饿再投喂组鱼其脑中表达量会显著升高,随后会有下降趋势,表明mc5r可能与光倒刺鲃的摄食调控有关。  相似文献   

6.
运用同源克隆的方法获得岩原鲤(Procypris rabaudi(Tchang))肌肉生长抑制素(myostatin,MSTN)c DNA全长序列,并通过逆转录PCR(RT-PCR)检测MSTN基因的组织差异性表达以及饥饿胁迫对MSTN基因表达的影响。结果显示,岩原鲤MSTN基因1 547 bp的c DNA序列,编码区为1 128 bp,编码375个氨基酸。PCR检测显示,该基因在肌肉和脑组织中大量表达,在眼、肠、心、鳃、肾和头肾组织中少量表达,在肝胰脏、脾脏组织中未见表达。饥饿再投喂实验表明肌肉组织中MSTN基因表达含量随着饥饿时间的延长逐渐升高,恢复投喂后3d表达含量急剧下降,投饵后6 d升至正常水平。结果表明,MSTN的表达对营养摄入敏感,在饥饿状态下岩原鲤通过上调MSTM表达来抑制肌肉生长,节约能量消耗。  相似文献   

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8.
本文利用生物信息学软件分析了鲤(Cyprinus carpio)基因组中同源框基因Hoxa3a与多种鱼类及其它生物的同源性,构建了分子进化树。结果表明:鲤与斑马鱼(Danio rerio)的序列同源性最高为95%,与大西洋鲑(Salmo salar)等四种鱼类的同源性次之为84%,与米氏叶吻银鲛(Callorhinch...  相似文献   

9.
脂肪酸合成酶(FASN)是动物体内脂肪酸合成的关键酶。本研究利用逆转录PCR和RACE技术获得鲤Cyprinus carpio FASN全长cDNA序列为8 927bp,开放阅读框7 533bp,编码2 511个氨基酸。FASN蛋白质相对分子量274 145.67D,理论等电点(PI)为6.10。氨基酸同源性分析结果显示:鲤FASN基因与其他鱼类同源性为75.13%~95.34%,与人同源性为61.81%。系统进化树结果显示:鲤FASN氨基酸序列与金线鲃Sinocyclocheilus grahamia聚为一支,同源性最高。实时荧光定量PCR(RT-qPCR)检测结果表明:FASN基因鲤脑组织中表达量最高,肝脏次之,血液中最低。鲤FASN基因的获得为进一步深入研究鲤脂肪酸的合成途径及脂肪发育分子调控机制奠定了基础。  相似文献   

10.
本文利用生物信息学软件分析了鲤(Cyprinus carpio)基因组中同源框基因Hoxa3a与多种鱼类及其它生物的同源性,构建了分子进化树。结果表明:鲤与斑马鱼(Danio rerio)的序列同源性最高为95%,与大西洋鲑(Salmo salar)等四种鱼类的同源性次之为84%,与米氏叶吻银鲛(Callorhinchus milii)和鳐(Leucoraja erinacea)的同源性最低为74.5%,与三种鸟类的同源性为75%左右。这结果也说明,Hox同源框基因序列在进化上高度保守。在序列比较的同时发现了2个鲤的SNP位点,T101C和A213G。上述研究结果,对于保护生物多样性(尤其是遗传多样性)的研究,揭示生物进化历程及机理具有十分重要的意义。  相似文献   

11.
设置持续投喂组(C,持续投喂8周)、饥饿再投喂组(R,饥饿4周+再投喂4周)和持续饥饿组(S,饥饿8周)3个处理组,研究3种不同饥饿处理对草鱼(Ctenopharyngodon idellus)血清生化指标、糖原和糖代谢相关酶和葡萄糖转运蛋白1(GLUT1)的影响,同时在此实验基础上研究草鱼在急性高糖负荷胁迫下的糖耐受能力、糖代谢相关酶和GLUT1的变化规律,旨在阐明草鱼在饥饿及再投喂处理条件下的糖代谢特征。选取初重为(125.35±0.54)g的草鱼,饲养8周后以30 mg/100 g体重的剂量腹腔注射葡萄糖研究其糖耐受能力。结果显示,S组肝糖原和血清的血糖、甘油三酯含量均最低。饥饿处理对草鱼糖耐受能力影响显著,S组血糖含量在各时间点上显著低于其余两组(P0.05),肝糖原在6 h达到峰值;饥饿处理对草鱼肝脏糖代谢关键酶影响显著,饥饿处理(S组)诱使磷酸烯醇式丙酮酸激酶(PEPCK)活性上升但抑制丙酮酸激酶(PK)和果糖-6-磷酸激酶(PFK)的活性(P0.05),而饥饿再投喂(R组)后PEPCK、PK和PFK酶活性恢复到持续投喂(C组)处理水平。注射葡萄糖后S组肝脏GK酶活性增幅最大,PK酶活性呈上升趋势,而R组则呈先下降后上升的趋势;饥饿处理对草鱼肝脏和肌肉GLUT1表达影响显著,注射葡萄糖后,除R组肝脏组织外,其余各组草鱼肝脏和肌肉组织GLUT1表达量均呈先上升后下降的趋势,且S组肌肉GLUT1表达量在各个时间点上均高于其余两组(P0.05)。研究结果表明,在不同饥饿处理下,草鱼可通过消耗肝糖原和甘油三酯及降低肝脏糖酵解相关酶(PK和PFK)活性和促进糖异生PEPCK酶活性来应对饥饿胁迫,而饥饿处理可诱使GK和PK酶活性上升、促进糖原合成和激活GLUT1基因的表达和转运来缓解草鱼急性高糖负荷,从而提高其糖耐受能力。  相似文献   

12.
This study assessed the effect of starvation on survival and nutritional status of newborn juveniles H. erectus (<10 days) to optimize rearing protocols, thereby helping to reduce wildlife exploitation. Maximum starvation time (MST) was estimated through the survival of juveniles continuously starved from birth. Resistance to starvation and the effect of food re‐introduction after 1, 2, 4 and 6 days of starvation on survival and metabolite concentrations (total proteins, total lipids, acylglycerides, cholesterol, glucose) were also determined. Survival amongst continuously starved animals decreased from 6.6 ± 0.5 to 0% from days 9 to 10 of starvation. Seahorses under different starvation–refeeding treatments all had 100% survival up to day 5 of experiments. After 10 days, however, a 4‐day starvation period followed by refeeding showed negative effects with <50% survival. During continuous starvation, lipids were the first energy reserve used to maintain basal metabolism, followed by proteins. Except for cholesterol, all metabolite concentrations differed between continuous starvations and feeding. Despite high seahorse survival after 5 days in the absence of food, the recovery of the metabolic status is possible after a starvation period of no more than 2 days, since irreversible physiological changes compromising the ultimate survival of the organisms take place after this time.  相似文献   

13.
Here, we aimed to study the slow muscle of the fish Piaractus mesopotamicus submitted to 30 days of fasting (D30) followed by 1 day (D31) or 30 days of refeeding (D60). Histological analysis of fibre diameter was performed in D30 and D60. The gene expression of parvalbumin (pvalb), atrogenes (murf1a, murf1b, mafbx) and anabolic genes (igf‐1, mtor) was analysed using RT‐qPCR in D30, D31 and D60. The proteome was obtained by shotgun proteomics at D30 and D60, and the set of differentially expressed proteins was analysed by bioinformatics. In all experiments, the control was regularly fed fish. The histological analysis showed no changes in muscle fibre diameter. The expression of catabolic and anabolic genes was not changed, except for the downregulation of igf‐1 in D30 and of mafbx in D31. The expression of pvalb was not changed in D30 and D60 but was decreased in D31. The proteomic analysis identified 169 proteins in D30 (24 upregulated and 18 downregulated) and 170 proteins in D60 (17 upregulated and 21 downregulated); many of them were related to energetic metabolism and intracellular Ca2+ homoeostasis. Overall, our results indicate that the slow‐twitch muscle presented few changes upon prolonged fasting and refeeding condition.  相似文献   

14.
To increase the current knowledge about the relationship between nutritional status and the digestive capacity of Siberian sturgeon (Acipenser baerii), we addressed the effect of starvation‐refeeding and macronutrient composition on growth parameters and key digestive enzyme activities in A. baerii. Acipenser baerii juveniles were fed four different diets for 3 weeks, then starved for 2 weeks and allowed to refed for 5 weeks with the same diets. Another group of fish were fed 10 weeks with the corresponding diets. Among 10‐week fed fish, high‐protein diets promoted higher body weight values, while the lowest specific growth rate was observed in fish fed a low‐protein, medium‐carbohydrate, high‐lipid diet (p < .05). At the end of the experiment, in fish refed for 5 weeks following a feeding‐starvation cycle and in 10‐week fed animals, the higher levels of blood glucose, triglycerides and cholesterol were found in fish fed low‐protein diets (p < .05). In all treatments, 2 weeks of starvation decreased α‐amylase activity in the intestine (p < .05), while 4 days of refeeding increased lipase (p > .05) and α‐amylase activity in the intestine as well as pepsin in the stomach (p < .05). Our findings suggest that A. baerii maintains a high capacity to digest proteins and lipids after 2 weeks of starvation and that α‐amylase can be used as an indicator of the nutritional status in fish submitted to starvation‐refeeding cycles. Indeed, refeeding with high‐protein and CHO:L ratio diets after starvation could improve the growth rate of A. baerii in culture.  相似文献   

15.
We investigated the effect of dietary supplementation with a branched‐chain amino acid (BCAA) mixture (valine, leucine and isoleucine) on juvenile yellowtail (Seriola quinqueradiata) growth and hepatic insulin‐like growth factor (IGF) gene expression. Total dietary BCAA content was 8.1, 8.5, 9.0 and 9.8 g   kg?1 for the control, 0.4%, 1.0% and 2.0% BCAA experimental groups, respectively. Body weight was measured on day 77, after which fish were fasted for 3 days and refed; the livers were then removed 3 or 9 h later for analysis of hepatic gene expression. Body weight significantly increased in the BCAA‐supplemented groups. Hepatic expression of IGF1 and IGF2 significantly increased in the 2% BCAA group at 3 and 9 h after refeeding. In the 1% BCAA group, hepatic IGF1 expression tended to be higher at 9 h than in the control group after refeeding. Also, hepatic IGF2 significantly increased at only 9 h after refeeding in the 1% BCAA group. In conclusion, dietary supplementation with crystalline BCAAs increased growth and hepatic expression of IGF1 and IGF2. These results suggest that dietary crystalline BCAA supplementation would be a valuable addition to yellowtail aquaculture practices.  相似文献   

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17.
为了解2种状态(饥饿和复投喂)下投喂3种糖源对凡纳滨对虾仔虾生长、虾体组成成分、代谢指标的影响,实验共5个处理,分别为饥饿组S0、对照组C、实验组S1、S2、S3(在基础饲料中糖源分别为:葡萄糖、蔗糖、玉米淀粉)。实验选取体质量为(1.84±0.23) g的凡纳滨对虾仔虾用方形纱制网兜独立喂养,进行为期12 d的饥饿实验后继续复投喂12 d。结果显示,饥饿后仔虾体组成成分及相关酶[脂肪酶(LPS)、磷酸果糖激酶(PFK)、已糖激酶(HK)、谷氨酰胺合成酶(GS)]差异显著;仔虾肝糖原、肌糖原均呈现出反复升降的过程,饥饿8 d后肝糖原降到最低值,肌糖原短暂回升后显著下降。复投喂4 d后S3组增重率最高,实验各组间无显著差异,均低于C组;全虾水分、全虾粗灰分、肌糖原含量无显著差异;S1、S2组虾体粗脂肪含量显著高于S3组;肝糖原、肌糖原均有回升,S1组肝糖原显著低于其他组。复投喂12 d后,S3组LPS活性、HK活性显著高于其他各实验组,GS、PFK含量实验组间无显著差异。研究表明,凡纳滨对虾仔虾饥饿过程中糖原和脂肪先于蛋白质被动用供能;复投喂出现部分补偿生长效应,玉米淀粉作为糖源饲料对凡纳滨对虾仔虾期恢复生长效果最佳。  相似文献   

18.
为促进肉食性鱼类人工配合饲料开发的理论基础研究,分析鱼类脂肪代谢的机制,实验克隆了大口黑鲈2个脂蛋白脂肪酶基因LPLtype1和LPLtype2的cDNA。序列分析表明,LPLtype1基因cDNA序列全长2 156 bp,编码516个氨基酸;LPLtype2基因cDNA序列全长1 710 bp,编码346个氨基酸。大口黑鲈LPLtype2与LPLtype1氨基酸序列之间的同源性为43.5%。系统进化分析表明,大口黑鲈LPLtype1和鳜LPL聚为一支,大口黑鲈LPLtype2和大麻哈鱼LPLtype2紧密聚为一支。预测分析发现,大口黑鲈LPLtype1和LPLtype2基因编码蛋白的活性中心位点、N-糖基化位点、二聚体形成的保守疏水残基位点、肝素结合域等主要功能域与硬骨鱼类和其他脊椎动物对比都比较保守。运用实时定量PCR方法检测脂蛋白脂肪酶mRNA的组织分布,发现LPLtype1和LPLtype2都在肝脏中表达量最高,推测这与肝脏是最主要的营养诱导性储脂部位有关。  相似文献   

19.
Chinese perch, an obligate piscivorous fish, could serve as an excellent model system for studies on the diversification of lipoprotein lipase (Lpl) and hepatic lipase (Hl) genes. In this study, we characterized the genomic structure, tissue expression and single nucleotide polymorphisms of Lpl and Hl genes in Chinese perch (Siniperca chuatsi). Consistent with findings in other vertebrates, Chinese perch Lpl gene consisted of ten exons and nine introns, and Hl gene consisted of nine exons and eight introns. The promoter region of Hl gene was not highly homologous to that of Lpl gene in Chinese perch. The mRNA expression of Lpl in Chinese perch was the highest in adipose tissue, followed by liver, brain, intestine and muscle, and the lowest in spleen, whereas Hl was almost exclusively expressed in liver. These results suggested that Chinese perch Lpl and Hl might be derived from an early duplication of an ancestral gene. Polymorphisms of Lpl and Hl genes were examined in feeders and non‐feeders of dead prey fish by direct sequencing of these genes in 130 fish from each group. Three SNPs (A1220T, G1221T and C1224G) were identified in Lpl gene, whereas none was identified in Hl gene. Diplotype 2 in Lpl gene was significantly correlated with acceptance of dead prey fish.  相似文献   

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