Essential role of NKCC1 in NGF-induced neurite outgrowth
Essential role of NKCC1 in NGF-induced neurite outgrowth
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DOI：
10.1016/j.bbrc.2007.05.133
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发表时间：
2007-08-03
影响因子：
3.1
通讯作者：
Marunaka, Yoshinori
中科院分区：
文献类型：
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作者：
Nakajima, Ken-ichi;Miyazaki, Hiroaki;Marunaka, Yoshinori
Na+/K+/2Cl(-)共转运体(NKCC)介导2Cl(-)与Na+、K+通过质膜的电子中和转运，在细胞摄取氯离子、细胞内氯离子的动态平衡和细胞体积调节中起着重要作用。然而，关于离子转运体在神经细胞突起生长中的作用，我们的信息非常有限。在本研究中，我们报道了NKCC1(NKCC的一个亚型)在NGF诱导的大鼠嗜铬细胞瘤PC12D细胞突起生长中的作用。经NGF处理后，NKCC1蛋白表达水平升高。RNA干扰(RNAi)下调NKCC1基因可显著抑制NGF诱导的神经突起生长。将增强型绿色荧光蛋白(EGFP)标记的大鼠NKCC1基因导入细胞，以明确NKCC1的细胞内定位，结果表明，在神经突起生长过程中，EGFP-rNKCC1主要定位于生长锥的质膜中。这些观察结果表明，NKCC1在NGF诱导的PC12D细胞突起生长中起重要作用。(C)2007 Elsevier Inc.保留所有权利。
The Na+/K+/2Cl(-) cotransporter (NKCC) mediates electroneutral transport of 2Cl(-) coupled with Na+ and K+ across the plasma membrane, and plays crucial roles in Cl- uptake into the cells, homeostasis of cellular Cl-, and cell volume regulation. However, we have very limited information on the roles of ion transporters in neurite outgrowth in neuronal cells. In the present study, we report the role of NKCCl (an isoform, of NKCC) in NGF-induced neurite outgrowth of rat pheochromocytoma PC12D cells. The expression level of NKCCl protein was increased by NGF treatment. Knock-down of NKCCl by RNA interference (RNAi) drastically diminished the NGF-induced neurite outgrowth. Transfection of enhanced green fluorescent protein (EGFP)-tagged rat NKCCl into cells for clarification of intracellular localization of NKCCl revealed that the EGFP-rNKCCl was mainly localized in the plasma membrane at growth cone during neurite outgrowth. These observations suggest that NKCCl plays a fundamental role in NGF-induced neurite outgrowth of PC12D cells. (c) 2007 Elsevier Inc. All rights reserved.