鲤鱼（Cyprinus carpio）促性腺激素的纯化及其α亚基新转录本的发现
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摘要
促性腺激素(GonadotropinHormone,GtH)是由脊椎动物垂体前叶细胞合成与分泌的一类糖蛋白激素,主要生理作用是刺激生殖细胞的生长、发育、成熟、排精及排卵。鱼类中也存在两种典型的GtHs,即GtHI和GtHII,分别对应于哺乳类的卵泡刺激素(Follicle-Stimulating Hormone, FSH)和促黄体生成素(LuteinizingHormone,LH)。GtH是一个二聚体分子,由一个共同的α亚基和一个具有激素生物学特异性的β亚基组成,只有两个亚基结合在一起才能发挥激素的生物学活性。硬骨鱼的生殖发育虽然受很多因素的影响,但由垂体前叶细胞和性腺介质细胞(folliclecell)分泌的GtH在性腺发育和性成熟中起主要调控作用。
鲤鱼(common carp, Cyprinus carpio)是我国常见的经济淡水鱼,也是常用于研究淡水鱼类内分泌调控的模式鱼。本研究室从上世纪80年代开始转基因鱼研究,目前已经研制成功了快速生长的转全鱼生长激素基因黄河鲤家系。通过转入生长激素基因,使该特定功能基因与基因组表达产物协调作用使鱼的生长速度加快从而缩短了生长周期。然而激素的调控是一个复杂的网络系统,一种激素的改变可能会引起另一种激素的变化。本实验室最近发现,转全鱼生长激素基因鲤鱼的第一次性成熟时间明显晚于对照鱼(本实验室待发表资料)。因此,转全鱼生长激素基因鲤鱼除具备优良养殖性状外,还有可能成为一种独特的模型,在鱼类内分泌学研究领域发挥作用。在硬骨鱼类,下丘脑合成的促性腺激素释放激素(Gonadotropin-releasing hormone, GnRH)直接通过神经轴突末梢作用于垂体前叶的促性腺激素细胞,与膜上分布的受体结合后刺激垂体合成的释放促性腺激素(GonadotropinHormone,GtH),进而激活性类固醇激素的生成,调节性腺发育和最后的性成熟,即性腺发育与性成熟的整个过程受下丘脑-垂体-性腺轴(hypothalamic-pituitary-Gonad, HPG)这个完整而协调的神经内分泌系统调控。本文就与性腺发育关系最密切的激素--促性腺激素,也是下丘脑-垂体-性腺轴的中间信号分子进行了以下方面的研究:
1.采用逆转录-聚合酶链式反应(RT-PCR)方法,从鲤鱼脑垂体获得GtHα和两
Gonadotropin (GtH) is a pituitary glycoprotein hormone that regulates gonaddevelopment and sex maturation in vertebrates. In teleost fish, as in other vertebrates,there are two forms of GtHs, traditionally referred to as GtH-I and GtH-II, which arebelieved to be FSH-like and LH-like, respectively. GtHs are dimeric glycoproteinhormones composed of a common alpha subunit and a hormone-specificβsubunit,which confers its biological specificity. The hormonal activity is expressed only aftera non-covalent association between these two subunits. Though many factors affectthereproductionanddevelopmentofteleostfish,GtHsecretedbythepituitaryplaysacrucial role in regulating gonad development and controlling the final sex maturationaswellasendocrineregulation.
Common carp (Cyprinus carpio), a familiar economic freshwater fish in China,isusedasthemodel organism ofendocrineregulationresearchoffreshwaterfish.Butthe molecular mechanism of regulating gonad development and reproduction ofcommon carp hasn’t been clearly elucidated. Since 1980s, transgenic carp has beenbred in our laboratory. Up to now fast-growing autotransgenic fish by transferringgrowth hormone gene into fish genome have been successfully obtained. The GHtransgenic carp has the feature of accelerated growth rate and shorten the time whichis required to reach the marketable size. However, Hormone regulation is a complexnetwork system, and the change of one hormone certainly will lead to the change ofanother one. Moreover, the sexural maturity of fast-growth GH transgenic commoncarp is highly delayed compared to the control ones (unpublished data). Therefore, inaddition to the desired traits for aquaculture, growth hormone transgenic commoncarp might become a special model which will play a role in the endocrinologystudyof fish. The present study focused on the GtH, the key signal molecular in theHypothalamic-Pituitary-Gonad (HPG) axis, which primarily stimulated gonaddevelopment.Themainresultsarepresentedasfollows:
鲤鱼(common carp, Cyprinus carpio)是我国常见的经济淡水鱼,也是常用于研究淡水鱼类内分泌调控的模式鱼。本研究室从上世纪80年代开始转基因鱼研究,目前已经研制成功了快速生长的转全鱼生长激素基因黄河鲤家系。通过转入生长激素基因,使该特定功能基因与基因组表达产物协调作用使鱼的生长速度加快从而缩短了生长周期。然而激素的调控是一个复杂的网络系统,一种激素的改变可能会引起另一种激素的变化。本实验室最近发现,转全鱼生长激素基因鲤鱼的第一次性成熟时间明显晚于对照鱼(本实验室待发表资料)。因此,转全鱼生长激素基因鲤鱼除具备优良养殖性状外,还有可能成为一种独特的模型,在鱼类内分泌学研究领域发挥作用。在硬骨鱼类,下丘脑合成的促性腺激素释放激素(Gonadotropin-releasing hormone, GnRH)直接通过神经轴突末梢作用于垂体前叶的促性腺激素细胞,与膜上分布的受体结合后刺激垂体合成的释放促性腺激素(GonadotropinHormone,GtH),进而激活性类固醇激素的生成,调节性腺发育和最后的性成熟,即性腺发育与性成熟的整个过程受下丘脑-垂体-性腺轴(hypothalamic-pituitary-Gonad, HPG)这个完整而协调的神经内分泌系统调控。本文就与性腺发育关系最密切的激素--促性腺激素,也是下丘脑-垂体-性腺轴的中间信号分子进行了以下方面的研究:
1.采用逆转录-聚合酶链式反应(RT-PCR)方法,从鲤鱼脑垂体获得GtHα和两
Gonadotropin (GtH) is a pituitary glycoprotein hormone that regulates gonaddevelopment and sex maturation in vertebrates. In teleost fish, as in other vertebrates,there are two forms of GtHs, traditionally referred to as GtH-I and GtH-II, which arebelieved to be FSH-like and LH-like, respectively. GtHs are dimeric glycoproteinhormones composed of a common alpha subunit and a hormone-specificβsubunit,which confers its biological specificity. The hormonal activity is expressed only aftera non-covalent association between these two subunits. Though many factors affectthereproductionanddevelopmentofteleostfish,GtHsecretedbythepituitaryplaysacrucial role in regulating gonad development and controlling the final sex maturationaswellasendocrineregulation.
Common carp (Cyprinus carpio), a familiar economic freshwater fish in China,isusedasthemodel organism ofendocrineregulationresearchoffreshwaterfish.Butthe molecular mechanism of regulating gonad development and reproduction ofcommon carp hasn’t been clearly elucidated. Since 1980s, transgenic carp has beenbred in our laboratory. Up to now fast-growing autotransgenic fish by transferringgrowth hormone gene into fish genome have been successfully obtained. The GHtransgenic carp has the feature of accelerated growth rate and shorten the time whichis required to reach the marketable size. However, Hormone regulation is a complexnetwork system, and the change of one hormone certainly will lead to the change ofanother one. Moreover, the sexural maturity of fast-growth GH transgenic commoncarp is highly delayed compared to the control ones (unpublished data). Therefore, inaddition to the desired traits for aquaculture, growth hormone transgenic commoncarp might become a special model which will play a role in the endocrinologystudyof fish. The present study focused on the GtH, the key signal molecular in theHypothalamic-Pituitary-Gonad (HPG) axis, which primarily stimulated gonaddevelopment.Themainresultsarepresentedasfollows:
引文
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