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Alternative TitleGrowth Hormone impacts on the gonadal development of all-fish GH transgenic carp
Thesis Advisor胡炜
Degree Grantor中国科学院水生生物研究所
Place of Conferral水生生物研究所
KeywordHpg轴 Gh/igf轴 转全鱼gh基因鲤 实时荧光定量rt-pcr 基因表达
Abstract鱼类性腺的发育主要受到下丘脑-垂体-性腺(hypothalamic-pituitary-gonad,HPG)轴的调控。同时,越来越多的研究表明生长激素/胰岛素类生长因子(Growth Hormone/ Insulin-like growth factor, GH/IGF)轴对鱼类性腺的发育也发挥着不可忽视的作用。 为了探索GH/IGF轴对鱼类性腺发育的作用机制,阐明HPG轴和GH/IGF轴之间相互作用的关系,本论文以对照鲤鱼胚胎为材料,采用RT-PCR的方法,研究了鲤鱼HPG轴和GH/IGF轴中几个关键信号分子的起始表达特性,结果显示大部分基因于受精后20h和23h开始表达;GHR和IGF-1均早于GH开始表达;GtH β亚基和α亚基的初始表达时间不同步。与此同时,本论文以过量表达GH的转全鱼GH基因鲤为模型,利用实时定量RT-PCR技术,分析了HPG轴以及GH/IGF轴中重要信号分子的表达规律。结果发现,在转基因鱼和对照鱼的垂体中GtH-I β亚基、GtH-II β亚基和GH以及肝脏中GHR和IGF-1的表达有明显的季节性,在春季性成熟时期表达有所上升;同时,发现了对照鲤鱼性腺中存在GHR的表达,为阐明GH在性腺中直接发挥作用提供了科学参数;另外,通过对转基因鱼和对照鱼的比较分析发现,转基因鱼垂体中GtH-I β亚基的表达受到了负向调控,肝脏中GHR的表达受到正向调控,而肝脏中IGF-1的表达则表现出了年龄差异:一龄鱼对照鱼垂体中IGF-1的相对表达量高于转基因鱼,而二龄鱼正好相反;肝脏中,GHR和IGF-1的表达存在着关联性。通过对以上几个关键信号分子的研究发现,GH的过量表达对HPG轴以及GH/IGF轴产生了重要影响,表明GH在鲤鱼性腺发育中发挥着不容忽视的调控作用。
Other AbstractConsidered as the pivotal axis in the regulation of sexual maturation, Hypothalamic-Pituitary-Gonad(HPG) axis has been studied thoroughly in the teleosts. In addition, there are evidences showing that Growth Hormone/Insulin-like growth factor(GH/IGF) axis also involve in the process of gonad development. For a better understanding of the relationship between the HPG axis and GH/IGF axis and the function of GH/IGF axis in the regulation of gonad development, the expression of sGnRH, cGnRH, GtH α subunit, GtH-I β subunit, GtH-II β subunit, GH, growth hormone receptor (GHR), IGF-1 were detected in carp embryos. Development in 20 hours post fertilization (hpf) and 23hpf was critical for carp embryos, as most of genes analyzed were first detectable at 20hpf or 23hpf. GHR and IGF-1 were all first expressed prior to GH. The differences in the timing of the expression in GH and IGF mRNA may suggest that IGF gene expression is not GH-dependent. Additionally, the timing of the expression of GtH α subunit, GtH-I β subunit, GtH-II β subunit were unsynchronized in embryos. At the same time, non-transgenic (wild-type) carp and growth hormone transgenic carp were examined for pituitary, liver and gonad mRNA levels. GtH-I β subunit, GtH-II β subunit, GH mRNA levels in the pituitary and GHR, Insulin-like growth factor-1(IGF-1) mRNA levels in the liver show a clear seasonal pattern, increasing in spring during sexual maturation. GHR mRNA levels in liver were higher in transgenic than non-transgenic carp, while GtH-I βsubunit mRNA levels in pituitary were higher in non-transgenic than transgenic carp. These data reveal the stimulation effects of GH in the expression of GHR in liver and the feedback regulation of GtH-I βsubunit production in the presence of constitutive GH overexpression in transgenic carp. In addition, the expression pattern of IGF-1 shows differences between two age groups: IGF-1 mRNA levels in liver of one-year old carp were higher in non-transgenic than transgenic carp. Contrarily, in two-year old carp IGF-1 mRNA levels in liver were higher in transgenic than non-transgenic carp. What’s more, GHR expression was identified in gonad, suggesting the possibility of direct action of GH in gonad. In liver, there was a relationship between the expression of GHR and IGF-1. These results suggest that GH involves in the process of gonad development by the regulation of HPG axis and GH/IGF axis.
Document Type学位论文
Recommended Citation
GB/T 7714
卢月娇. 生长激素基因对转基因黄河鲤性腺发育影响的初步研究[D]. 水生生物研究所. 中国科学院水生生物研究所,2008.
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