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欧洲鳗鲡鸟苷素的克隆与表达

Cloning and Expression of Guanylin from the European eel (Anguilla anguilla).

作者信息

Comrie M M, Cutler C P, Cramb G

机构信息

School of Biology, University of St. Andrews, Bute Medical Buildings, St. Andrews, Fife, United Kingdom.

出版信息

Biochem Biophys Res Commun. 2001 Mar;281(5):1078-85. doi: 10.1006/bbrc.2001.4485.

Abstract

Extracts of intestinal epithelia from the European eel (Anguilla anguilla) stimulated cGMP production in the T84 human colon carcinoma cell line which suggested the presence of a guanylin-like peptide in this teleost fish. Degenerate oligonucleotide primers were subsequently used in RT-PCR resulting in the amplification, cloning, and sequencing of two cDNAs which represent possible 5' spliceoforms of an eel homologue of the mammalian peptide, guanylin. Northern blotting indicated that the main site of expression of the eel peptide is in the intestine with much lower signals also detected in the kidney. Intestinal expression of guanylin mRNA is up-regulated in both nonmigratory "yellow" and the more sexually mature, migratory "silver" eels following acclimation to the seawater environment. These results suggest that this peptide signalling system may play a role in osmoregulation in euryhaline teleost fish during migration between the marine and freshwater environments.

摘要

欧洲鳗鲡(Anguilla anguilla)肠道上皮提取物可刺激T84人结肠癌细胞系中cGMP的产生,这表明这种硬骨鱼中存在一种类鸟苷蛋白肽。随后,简并寡核苷酸引物被用于逆转录聚合酶链反应(RT-PCR),从而扩增、克隆并测序了两个互补DNA(cDNA),它们代表了哺乳动物肽鸟苷蛋白的鳗鱼同源物可能的5'剪接异构体。Northern印迹法表明,鳗鱼肽的主要表达部位是肠道,在肾脏中也检测到较低的信号。在适应海水环境后,非洄游的“黄色”鳗鱼和性成熟度更高的洄游“银色”鳗鱼的肠道中鸟苷蛋白mRNA的表达均上调。这些结果表明,这种肽信号系统可能在广盐性硬骨鱼在海洋和淡水环境之间洄游期间的渗透调节中发挥作用。

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