Inhibition of protocadherin-α function results in neuronal death in the developing zebrafish
Inhibition of protocadherin-α function results in neuronal death in the developing zebrafish
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DOI：
10.1016/j.ydbio.2008.06.011
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发表时间：
2008-09-01
影响因子：
2.7
通讯作者：
Jontes, James D.
中科院分区：
文献类型：
--
作者：
Emond, Michelle R.;Jontes, James D.
pcdh α /CNR基因由一系列钙粘蛋白超家族的神经细胞表面蛋白组成，尽管人们对它们在神经发育中的作用知之甚少。在这里，我们提供了斑马鱼中pcdhl α的第一个深入表征。全挂载免疫细胞化学表明，很大一部分内源性胞质结构域的免疫反应性存在于细胞核中，这表明细胞内结构域的内溶蛋白裂解和核易位是体内Pcdh1 α活性的重要方面。利用全挂载免疫细胞化学和基于bacc的Pcdh1 α - gfp融合蛋白的表达，我们发现Pcdh1 α似乎没有形成稳定的，突触发生早期的突触点。我们还证明了Pcdhl α细胞质结构域的存在对于正常功能是必不可少的。截断Pcdhl α蛋白，使用剪接阻断反义morpholinos阻止整个Pcdhl α簇的共同胞内结构域的添加，导致整个发育中的脑和脊髓的神经元凋亡，表明Pcdhl α在早期神经发育中的重要作用。这种细胞死亡表型可以通过可溶性Pcdhl α细胞质结构域的表达而减弱。(C) 2008爱思唯尔公司版权所有。
The pcdh alpha/CNR gene comprises a diverse array of neuronal cell-surface proteins of the cadherin superfamily, although very little is known about their role in neural development. Here we provide the first in-depth characterization of pcdhl alpha in zebrafish. Whole-mount immunocytochemistry demonstrates that a large proportion of endogenous Cytoplasmic domain immunoreactivity is present in the nucleus, suggesting that endoproteolytic cleavage and nuclear translocation of the intracellular domain are important aspects of pcdhl alpha activity in vivo, Using whole-mount immunocytochemistry and BAC-based expression of Pcdh1 alpha-GFP fusion proteins,we find that Pcdhl alpha does not appear to form stable, synaptic puncta at early stages of synaptogenesis. We also demonstrate that the presence of the Pcdhl alpha cytoplasmic domain is essential for normal function. Truncation of Pcdhl alpha proteins, Using splice-blocking antisense morpholinos to prevent the addition of the common intracellular domain to the entire pcdhl alpha cluster, results in neuronal apoptosis throughout the developing brain and spinal cord, demonstrating an essential role for pcdhl alpha in early neural development.This cell death phenotype can be attenuated by the expression of a soluble Pcdhl alpha cytoplasmic domain. (C) 2008 Elsevier Inc. All rights reserved.