文章摘要
樊佳佳,白俊杰,李小慧,杜芳芳,于凌云,叶星.大口黑鲈生长激素促分泌素cDNA结构和早期发育阶段表达谱分析[J].水产学报,2010,34(11):1656~1663
大口黑鲈生长激素促分泌素cDNA结构和早期发育阶段表达谱分析
Ghrelin cDNA structure and expression profile in early development of largemouth bass(Micropterus salmoides)
投稿时间:2010-02-05  修订日期:2010-08-16
DOI:10.3724/SP.J.1231.2010.06834
中文关键词: 大口黑鲈  生长激素促分泌素  实时定量PCR  早期发育  表达谱
英文关键词: largemouth bass(Micropterus salmoides)  ghrelin  real-time PCR  early development  expression profile
基金项目:国家科技支撑计划项目(2006BAD01A1209);广东省科技计划项目(2007B020708008);农业部公益性行业科研专项(200903045);国家自然科学基金项目(30600152)
作者单位E-mail
樊佳佳 中国水产科学研究院珠江水产研究所,中国水产科学院热带亚热带鱼类选育与养殖重点开放实验室  
白俊杰 中国水产科学研究院珠江水产研究所,中国水产科学院热带亚热带鱼类选育与养殖重点开放实验室 jjbai@163.net 
李小慧 中国水产科学研究院珠江水产研究所,中国水产科学院热带亚热带鱼类选育与养殖重点开放实验室  
杜芳芳 中国水产科学研究院珠江水产研究所,中国水产科学院热带亚热带鱼类选育与养殖重点开放实验室  
于凌云 中国水产科学研究院珠江水产研究所,中国水产科学院热带亚热带鱼类选育与养殖重点开放实验室  
叶星 中国水产科学研究院珠江水产研究所,中国水产科学院热带亚热带鱼类选育与养殖重点开放实验室  
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中文摘要:
      研究采用RT-PCR、RACE和PCR技术从大口黑鲈胃组织中克隆得到ghrelin基因的cDNA。该基因全长434 bp,其中开放阅读框(open reading frame,ORF)321bp,编码107个氨基酸的前肽,20个氨基酸的成熟肽位于第27位到第46位氨基酸处。前肽和成熟肽的氨基酸与已报导的舌齿鲈进行同源性分析,相似性分别为85%和90%。为进一步了解ghrelin基因在鱼体早期发育阶段的表达,本实验采用实时定量PCR(real-time PCR)方法检测了ghrelin基因在大口黑鲈胚胎和仔鱼发育过程的表达谱。结果显示,ghrelin基因在受精卵时期就有少量表达,但直到体节出现之前表达量均较低。出膜第4天ghrelin基因出现大量表达,第12天ghrelin基因表达量更显著增加,出膜第4天和第12天的表达量分别是受精卵时期表达量的206.77倍和531.20倍。出膜第4天正是大口黑鲈仔鱼“开口觅食期”,仔鱼消化系统初步发育成型,由完全利用卵黄囊营养转为从外界觅食阶段,到出膜第12天,仔鱼消化系统已发育完善,完全依靠外界营养提供能量,由ghrelin基因的表达量变化可推测其可能参与了鱼类早期发育阶段的摄食调节。
英文摘要:
      Ghrelin,the endogenous ligand of growth hormone secretagogue receptor(GHS R),has been shown to play an important role in eliciting growth hormone release and energy homeostasis.In the present study,we cloned ghrelin from stomach tissue of largemouth bass(Micropterus salmoides)by using RT-PCR,RACE and PCR method.The amplified cDNA was totally 434bp in length including 321bp open reading frame(ORF).It encoded a propeptide of 107aa,with the predicted mature peptide consisting of 20aa.Alignment analyses showed that the similarities of propeptide and mature peptide beween Dicentrarchus labrax were 85% and 90% respectively.To better understand the expression during fish early development of ghrelin,we also detected the expression profile of ghrelin at embryo and early fry of largemouth bass by real-time PCR.The result showed that ghrelin mRNA was observed at fertilized egg stage,and increased at metamere stage.The expression level at 4-days and 12 days larvae was about 206.77 and 531.20 times higher than that at fertilized egg stage.The 4-days larva was “first feeding” in largemouth bass,when the larva was from endogenous nutrition to exogenous nutrition phase.The 12 day larvae had greatly increased intake appetite and at this stage larvae took full advantage of the exogenous nutrition,and the expression of ghrelin also reached the highest value.These results combined suggest that ghrelin is involved in the regulation of food intake in largemouth bass.
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