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小鼠伴侣蛋白t-复合体蛋白-1的蝾螈同源物在早期发育过程中的表达。

Expression of the axolotl homologue of mouse chaperonin t-complex protein-1 during early development.

作者信息

Sun H B, Neff A W, Mescher A L, Malacinski G M

机构信息

Department of Biology, Indiana University, Bloomington 47405.

出版信息

Biochim Biophys Acta. 1995 Jan 25;1260(2):157-66. doi: 10.1016/0167-4781(94)00187-8.

Abstract

Molecular chaperones assist in the folding of proteins, but their role during development is not well understood. Here we report the temporal and spatial expression pattern of the axolotl homologue of mouse chaperonin TCP-1 during normal amphibian embryogenesis and in several models of abnormal embryogenesis. A partial axolotl TCP-1 cDNA (646 bp; 519 coding bp) isolated by 3' RACE PCR shows considerable homology to mouse TCP-1. Developmental Northerns and RT-PCR analyses of whole axolot1 embryos revealed a low level of maternal TCP-1 transcripts in fertilized eggs. The maternal transcripts were down-regulated to a non-detectable level in early gastrulae. Zygotic TCP-1 transcripts first appeared during gastrulation. They were mainly expressed in mid-neurula and later stage embryos. Whole-mount in situ hybridization studies showed abundant TCP-1 transcripts in the blastopore at the mid-gastrula stage and in the brain and spinal cord beginning at the neurula stage, and in the somites (myotomes) at the tailbud stage. RT-PCR analysis of TCP-1 expression in axolotl embryos treated with either high salt (causing exogastrulation) or ultraviolet (UV) irradiation (causing ventralization) substantiated the correlation between TCP-1 expression and neural and somitic development. In high salt-induced exogastrulated embryos TCP-1 mRNA was detectable in the ectoderm part (with neural tissues) but not in its exogastrulated endoderm part. Lower levels of TCP-1 expression were detected in UV-irradiated, ventralized embryos with smaller head and reduced neural and somitic tissues. Normal levels of TCP-1 expression were detected in embryos with double axes/heads. These studies provide strong evidence that at the transcript level axolotl chaperonin TCP-1 is regulated both temporally and spatially during embryogenesis, especially in neural and somitic development.

摘要

分子伴侣协助蛋白质折叠,但其在发育过程中的作用尚未得到充分了解。在此,我们报告了小鼠伴侣蛋白TCP-1的蝾螈同源物在正常两栖动物胚胎发育过程中以及几种异常胚胎发育模型中的时空表达模式。通过3' RACE PCR分离得到的部分蝾螈TCP-1 cDNA(646 bp;519个编码bp)与小鼠TCP-1具有相当高的同源性。对整个蝾螈胚胎进行的发育Northern杂交和RT-PCR分析显示,受精卵中存在低水平的母体TCP-1转录本。母体转录本在早期原肠胚中下调至无法检测的水平。合子TCP-1转录本在原肠胚形成期间首次出现。它们主要在神经胚中期和后期胚胎中表达。全胚胎原位杂交研究表明,在原肠胚中期的胚孔、神经胚期开始的脑和脊髓以及尾芽期的体节(肌节)中存在丰富的TCP-1转录本。对用高盐(导致外胚层化)或紫外线(UV)照射(导致腹侧化)处理的蝾螈胚胎中TCP-1表达的RT-PCR分析证实了TCP-1表达与神经和体节发育之间的相关性。在高盐诱导的外胚层化胚胎中,TCP-1 mRNA在外胚层部分(含神经组织)中可检测到,但在其外胚层化的内胚层部分中未检测到。在头部较小且神经和体节组织减少的紫外线照射腹侧化胚胎中检测到较低水平的TCP-1表达。在具有双轴/双头的胚胎中检测到正常水平的TCP-1表达。这些研究提供了强有力的证据,表明在转录水平上,蝾螈伴侣蛋白TCP-1在胚胎发育过程中受到时空调控,尤其是在神经和体节发育中。

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