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Abu Shufian Ishtiaq Ahmed, 于力群, 朱作言, 孙永华. 生长激素受体信号相关因子在斑马鱼中的比较表达分析及GHR 信号通路体内研究模型的建立[J]. 水生生物学报, 2011, 35(5): 727-738. DOI: 10.3724/SP.J.1035.2011.00727
引用本文: Abu Shufian Ishtiaq Ahmed, 于力群, 朱作言, 孙永华. 生长激素受体信号相关因子在斑马鱼中的比较表达分析及GHR 信号通路体内研究模型的建立[J]. 水生生物学报, 2011, 35(5): 727-738. DOI: 10.3724/SP.J.1035.2011.00727
Abu Shufian Ishtiaq Ahmed, YU Li-Qun, ZHU Zuo-Yan, SUN Yong-Hua. COMPARATIVE EXPRESSION ANALYSIS OF GHR SIGNALING RELATED FACTORS IN ZEBRAFISH (DANIO RERIO) AND AN IN VIVO MODEL TO STUDY GHR SIGNALING[J]. ACTA HYDROBIOLOGICA SINICA, 2011, 35(5): 727-738. DOI: 10.3724/SP.J.1035.2011.00727
Citation: Abu Shufian Ishtiaq Ahmed, YU Li-Qun, ZHU Zuo-Yan, SUN Yong-Hua. COMPARATIVE EXPRESSION ANALYSIS OF GHR SIGNALING RELATED FACTORS IN ZEBRAFISH (DANIO RERIO) AND AN IN VIVO MODEL TO STUDY GHR SIGNALING[J]. ACTA HYDROBIOLOGICA SINICA, 2011, 35(5): 727-738. DOI: 10.3724/SP.J.1035.2011.00727

生长激素受体信号相关因子在斑马鱼中的比较表达分析及GHR 信号通路体内研究模型的建立

COMPARATIVE EXPRESSION ANALYSIS OF GHR SIGNALING RELATED FACTORS IN ZEBRAFISH (DANIO RERIO) AND AN IN VIVO MODEL TO STUDY GHR SIGNALING

  • 摘要: 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、stat5.1、stat5.2、igf1、c-fos、socs1 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igf1 转录活性以及GH 应激启动子spi2.1 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1dpf 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。

     

    Abstract: 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, socs1 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, igf1 and c-fos. By analyzing the transcription level of c-fos and igf1 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 of gh or ghr overexpression.

     

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