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稀有鮈鲫卵黄蛋白原的分子克隆以及肾上腺素受体激动剂对其表达的影响
Alternative TitleMolecular Cloning of Vitellogenin Ao1 of Chinese Rare Minnow, Gobiocypris rarus and Effects of Adrenergic Agonists on Its Expression
马立文
Subtype硕士
Thesis Advisor殷战
2008-06-12
Degree Grantor中国科学院水生生物研究所
Place of Conferral水生生物研究所
Keyword稀有鮈鲫 卵黄蛋白原 17ß-雌二醇 肾上腺素受体激动剂
Abstract硬骨鱼的卵黄蛋白原(Vitellogenin,VTG)是一个由多个保守区域组成的磷脂蛋白大分子,一般认为它是受雌激素刺激而产生的卵黄蛋白(Yolk protein,YP)的前体。我们用17ß-雌二醇 (estradiol,E2)溶液浸泡稀有鮈鲫(Gobiocypris rarus),提取其肝脏RNA,构建了稀有鮈鲫肝脏cDNA文库,并且从中鉴定出5个编码卵黄蛋白原的克隆。利用从稀有鮈鲫肝脏cDNA文库中得到的4个VTG cDNA的部分序列信息以及cDNA末端快速扩增 (rapid-amplification of cDNA ends,RACE)技术,我们得到了稀有鮈鲫vtgAo1的全长序列。通过反转录聚合酶链式反应(RT-PCR),我们可以观察到17ß-雌二醇可以明显诱导稀有鮈鲫肝脏中vtgAo1的表达。一般认为,vtgAo1转录体主要在肝脏中表达。然而,在我们的实验中,我们在心脏——这个意想不到的地方也发现了一定程度的vtgAo1表达。通过RT-PCR和整体原位杂交(whole mount in situ hybridization)分析,我们证实了vtgAo在心脏中的表达。用α-肾上腺素受体激动剂去氧肾上腺素(phenylephrine,PE)和β-肾上腺素受体激动剂异丙肾上腺素(isoproterenol,ISO)处理稀有鮈鲫,表明心脏中vtgAo1的表达可受肾上腺素受体激动剂抑制。我们知道哺乳动物心肌细胞在代谢过程中能够卸载胞内多余的脂质,这是由哺乳类动物中的载脂蛋白B(apolipoproteinB,apoB)通过“反向甘油三酯转运(reverse triglyceride transportation)”的方式而进行的。我们所观察到的稀有鮈鲫这种硬骨鱼心脏能够表达卵黄蛋白原,提示了它可能具有与在哺乳类动物中同源蛋白apoB相类似的心脏保护作用。我们的实验结果表明:硬骨鱼体内卵黄蛋白原不仅可以通过雌激素诱导而产生,而且也可以由肾上腺素信号诱导途径所调节。
Other AbstractTeleost vitellogenins (VTGs) are large multidomain and traditionally considered as the estrogen responsive precursors of the major egg yolk proteins. We have identified 5 clones encoding VTGs, about 16% of the random EST clones from our constructed cDNA library from Chinese rare minnow liver tissue treated with 17ß-estradiol (E2). Full-length vtgAo1 has been obtained based on the sequence information of the 4 partial cDNA inserts by RACE. The inducibility of the vtgAo1 expression in liver by E2 has been confirmed by RT-PCR. The presence of vtgAo1 transcripts have been observed primarily in liver. However, a significant level of the vtgAo1 has been found in an unexpected location, heart, particularly in atrial cells by RT-PCR and whole mount in situ hybridization analyses. The vtgAo1 mRNA expression in heart can be suppressed by both α-adrenergic agonist, phenylephrine (PE) andβ-adrenergic agonist, isoproterenol (ISO). The cardiac VTG expression observed in present studies suggests its similar protection functions for unload surplus intracellular lipid in cardiomyocytes for “reverse triglyceride transportation” as the ones of the lipid transport proteins in mammals. The results also indicate that the production of teleost vtg in vivo can be regulated by not only estrogenic agents, but adrenergic signals as well.
Pages93
Language中文
Document Type学位论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/12360
Collection学位论文
Recommended Citation
GB/T 7714
马立文. 稀有鮈鲫卵黄蛋白原的分子克隆以及肾上腺素受体激动剂对其表达的影响[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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