水生生物学报
水生生物學報
수생생물학보
ACTA HYDROBIOLOGICA SINICA
2011年
5期
727-738
,共12页
Abu Shufian Ishtiaq Ahmed%于力群%朱作言%孙永华
Abu Shufian Ishtiaq Ahmed%于力群%硃作言%孫永華
Abu Shufian Ishtiaq Ahmed%우력군%주작언%손영화
生长激素%生长激素受体%GHR信号通路%斑马鱼%转录分析
生長激素%生長激素受體%GHR信號通路%斑馬魚%轉錄分析
생장격소%생장격소수체%GHR신호통로%반마어%전록분석
GH%GHR%GHR signaling%Zebrafish%Transcriptional analysis
GHR信号通路在动物出生后的生长中扮演着重要角色.实验利用斑马鱼模型研究了GHR信号相关基因在成体组织、胚胎发育以及幼体期的表达情况,这些基因包括gh、ghra、ghrb、jak2a、jak2b、statS.1、stat5.2、igf1、c-fos、socsl和socs2.值得关注的是,上述的大部分基因都存在母源性表达,且它们的合子表达均起始在体节早期之前.这说明在有功能性的脑垂体形成之前和完善的循环系统建立之前,GH及GH信号相关因子就已经存在于早期胚胎中,因此GH很可能是控制胚胎发育的一系列自分泌/旁分泌生长因子中的一员.同时,我们发现成体组织的SocS表达水平与GH信号靶基因igf1和c-fos的表达呈某种程度的负相关.我们利用实时定量PCR技术和荧光素酶分析技术,通过注射GH和GHR表达载体,在斑马鱼胚胎中分析了它们促进GH信号靶基因c-fos和igfl转录活性以及GH应激启动子spi12.1活性的能力.由此,研究利用斑马鱼胚胎建立一个体内研究模型来评估发育过程中的GH信号激活(GHSA).在受精后1天(dpf)和3dpf斑马鱼胚胎中,单独过表达gh或ghr均可以显著刺激GHSA,这表明在ldpf的斑马鱼胚胎中即存在功能性的GH和GHR蛋白表达,而这一时期是在功能性垂体的形成之前的.gh以及ghr的协同过表达则可以显著放大gh或ghr单独过表达的GHSA效果.
GHR信號通路在動物齣生後的生長中扮縯著重要角色.實驗利用斑馬魚模型研究瞭GHR信號相關基因在成體組織、胚胎髮育以及幼體期的錶達情況,這些基因包括gh、ghra、ghrb、jak2a、jak2b、statS.1、stat5.2、igf1、c-fos、socsl和socs2.值得關註的是,上述的大部分基因都存在母源性錶達,且它們的閤子錶達均起始在體節早期之前.這說明在有功能性的腦垂體形成之前和完善的循環繫統建立之前,GH及GH信號相關因子就已經存在于早期胚胎中,因此GH很可能是控製胚胎髮育的一繫列自分泌/徬分泌生長因子中的一員.同時,我們髮現成體組織的SocS錶達水平與GH信號靶基因igf1和c-fos的錶達呈某種程度的負相關.我們利用實時定量PCR技術和熒光素酶分析技術,通過註射GH和GHR錶達載體,在斑馬魚胚胎中分析瞭它們促進GH信號靶基因c-fos和igfl轉錄活性以及GH應激啟動子spi12.1活性的能力.由此,研究利用斑馬魚胚胎建立一箇體內研究模型來評估髮育過程中的GH信號激活(GHSA).在受精後1天(dpf)和3dpf斑馬魚胚胎中,單獨過錶達gh或ghr均可以顯著刺激GHSA,這錶明在ldpf的斑馬魚胚胎中即存在功能性的GH和GHR蛋白錶達,而這一時期是在功能性垂體的形成之前的.gh以及ghr的協同過錶達則可以顯著放大gh或ghr單獨過錶達的GHSA效果.
GHR신호통로재동물출생후적생장중분연착중요각색.실험이용반마어모형연구료GHR신호상관기인재성체조직、배태발육이급유체기적표체정황,저사기인포괄gh、ghra、ghrb、jak2a、jak2b、statS.1、stat5.2、igf1、c-fos、socsl화socs2.치득관주적시,상술적대부분기인도존재모원성표체,차타문적합자표체균기시재체절조기지전.저설명재유공능성적뇌수체형성지전화완선적순배계통건립지전,GH급GH신호상관인자취이경존재우조기배태중,인차GH흔가능시공제배태발육적일계렬자분비/방분비생장인자중적일원.동시,아문발현성체조직적SocS표체수평여GH신호파기인igf1화c-fos적표체정모충정도적부상관.아문이용실시정량PCR기술화형광소매분석기술,통과주사GH화GHR표체재체,재반마어배태중분석료타문촉진GH신호파기인c-fos화igfl전록활성이급GH응격계동자spi12.1활성적능력.유차,연구이용반마어배태건립일개체내연구모형래평고발육과정중적GH신호격활(GHSA).재수정후1천(dpf)화3dpf반마어배태중,단독과표체gh혹ghr균가이현저자격GHSA,저표명재ldpf적반마어배태중즉존재공능성적GH화GHR단백표체,이저일시기시재공능성수체적형성지전적.gh이급ghr적협동과표체칙가이현저방대gh혹ghr단독과표체적GHSA효과.
Growth hormone receptor (GHR) signaling pathway plays an important role in postnatal growth of animals.Although significant progress has been made in elucidating the signaling pathways activated by GHR in recent years,a comparative expression analysis of all the GHR signaling related genes and evaluation of GH-signal activation (GHSA)in an in vivo model still remain elusive.The zebrafish (Danio rerio) is an excellent model organism to study both developmental and physiological processes.In the present study,we comparatively analyzed the expression of GHR signal related genes,such as gh,ghra,ghrb,jak2a,jak2b,stat5.1,stat5.2,igf1,c-fos,socsl and socs2 in adult tissues,and during embryonic development by means of RT-PCR.Maternal expression was found in most of them and their zygotic expression mostly started from early-somite stage,far before the differentiation of pituitary somatotrophs and before the establishment of a functional circulatory system.In adults,the tissue distribution abundance of socs mRNA was negatively correlated with the GH-signal targets,igfl and c-f os.By analyzing the transcription level ofc-fos and igfl and the promoter activity of spi2.1,in zebrafish embryos injected with different constructs,i.e.,the expression constructs of gh and ghr,we established an in vivo model to evaluate GH-signal activation (GHSA) in early development of zebrafish embryos.Overexpression of gh or ghr alone in zebrafish embryos could significantly stimulate GHSA at 1 day post-fertilization (dpf) and 3 dpf,strongly indicating the existence of functional GH and GHR proteins in zebrafish embryos from 1 dpf,far prior to the formation of functional pituitary gland.A synergetic overexpression of gh and ghr could amplify the GHSA effects ofgh or ghr overexpression.