STRUCTURE AND DISTRIBUTION OF N-CADHERIN IN DEVELOPING ZEBRAFISH EMBRYOS - MORPHOGENETIC EFFECTS OF ECTOPIC OVER-EXPRESSION
STRUCTURE AND DISTRIBUTION OF N-CADHERIN IN DEVELOPING ZEBRAFISH EMBRYOS - MORPHOGENETIC EFFECTS OF ECTOPIC OVER-EXPRESSION
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DOI：
10.1002/aja.1002010204
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发表时间：
1994-10-01
影响因子：
2.5
通讯作者：
GEIGER, B
中科院分区：
文献类型：
--
作者：
BITZUR, S;KAM, Z;GEIGER, B
从斑马鱼胚胎cDNA文库中克隆了N-钙粘蛋白cDNA。该分子（ZN-钙粘蛋白）的推导的氨基酸序列的分析显示了高度的同源性，其他物种的N-钙粘蛋白，除了它的前序列是相当短的。然而，转染到中国仓鼠卵巢（CHO）细胞后，表达的蛋白是功能活性的，即参与钙依赖性细胞间相互作用。此外，异位过表达的ZN-cadherin，以下mRNA显微注射到2-4细胞胚胎，引起微聚集和不均匀的分离的深层细胞，导致畸形胚胎。发育北方和Western印迹分析表明，mRNA和蛋白质首先出现在原肠胚形成。原位杂交结果表明，ZN-cadherin mRNA最初存在于所有的深层细胞，后来成为限制各种上皮和神经组织。整装免疫染色表明，虽然ZN-钙粘蛋白已经存在于50%的外延，它成为与细胞连接只有4-5小时后。在发育体节中，Zn-钙粘蛋白表达显著但短暂。在上皮体节中检测到高水平的蛋白质，其表达明显下调伴随着肌发生的发生。(C)1994 Wiley-Liss，Inc.
N-cadherin cDNA was cloned from a zebrafish embryonic cDNA library. Analysis of the deduced amino acid sequence of this molecule (ZN-cadherin) revealed a high degree of homology to N-cadherins of other species, except that its pre-sequence is considerably shorter. Nevertheless, following transfection into chinese hamster ovary (CHO) cells, the expressed protein was functionally active, namely participated in calcium-dependent intercellular interactions. Moreover, ectopic over-expression of ZN-cadherin, following mRNA microinjection into 2-4 cell embryos, caused microaggregation and uneven segregation of deep cells, resulting in distorted embryos. Developmental Northern and Western blot analyses indicated that both the mRNA and the protein first appear at gastrulation. In-situ hybridization showed that ZN-cadherin mRNA was initially present in all deep cells, and later became restricted to various epithelial and neural tissues. Whole-mount immunostaining indicated that while ZN-cadherin was already present at 50% epiboly, it became associated with cell junctions only 4-5 h later. In developing somites ZN-cadherin expression was prominent but transient. High levels of the protein were detected in epithelial somites and its expression was apparently down regulated concomitantly with the onset of myogenesis. (C) 1994 Wiley-Liss, Inc.