Neuritogenesis and the nerve growth factor-induced differentiation of PC-12 cells requires annexin II-mediated plasmin generation
Neuritogenesis and the nerve growth factor-induced differentiation of PC-12 cells requires annexin II-mediated plasmin generation
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DOI：
10.1074/jbc.m106289200
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发表时间：
2001-12-28
影响因子：
4.8
通讯作者：
Hajjar, KA
中科院分区：
文献类型：
--
作者：
Jacovina, AT;Zhong, FM;Hajjar, KA
神经生长因子(NGF)诱导大鼠肾上腺嗜铬细胞瘤(PC-12)细胞分化的关键形态变化之一是轴突样突起的生长。越来越多的证据表明，这一过程可能依赖于纤溶酶，纤溶酶是一种丝氨酸蛋白酶，由尿激酶型纤溶酶原激活剂(u-PA)或组织纤溶酶原激活剂(t-PA)产生。我们实验室以前的工作已经确定Annexin II(ANN-II)是PLG和t-PA的共同受体，促进并定位细胞表面附近的纤溶酶生成。在目前的研究中，我们报告了在NGF处理的PC-12细胞中ANN-II蛋白和消息水平增加了3-9倍。消息稳定性和核连续分析表明，这种诱导发生在基因转录水平。在三维基质上和三维基质内的突起生长分析表明，针对ANN-H的多克隆和单抗以及反义ANN-II mRNA的过表达抑制了NGF诱导的PC-12细胞的分化。α(2)-纤溶酶抑制剂、针对t-PA和u-PA的抗体以及E-氨基己酸也会损害神经发生，E-氨基己酸是一种赖氨酸类似物，可以抑制PLG的激活和PLG与ANN-II的结合。纤溶酶生成分析显示，NGF处理的PC-12细胞的纤溶酶生成增加了2倍，这种增加可以被针对ANN-H尾部的多克隆抗体所阻断。根据这些数据，我们得出结论，ANN-II在转录水平上受到NGF的上调，ANN-II介导的纤溶酶的产生可能在分化的PC-12细胞的轴突发育过程中发挥重要作用。
One of the key morphological changes associated with the nerve growth factor (NGF)-induced differentiation of rat adrenal pheochromocytoma (PC-12) cells is the growth of axon-like processes called neurites. A growing body of evidence suggests that this process may be dependent upon plasmin, a serine protease generated from plasminogen (Plg) by either urokinase Plg activator (u-PA) or tissue Plg activator (t-PA). Prior work in our laboratory has identified annexin II (Ann-II) as a co-receptor for Plg and t-PA that promotes and localizes plasmin generation near the cell surface. In the present study, we report a 3-9-fold increase in Ann-II protein and message levels in NGF-treated PC-12 cells. Message stability and nuclear run-on assays suggest that this induction occurs at the level of gene transcription. Neurite outgrowth assays on and within a three-dimensional matrix demonstrate the inhibition of NGF-induced PC-12 cell differentiation by polyclonal and monoclonal antibodies directed against Ann-H as well as by the overexpression of antisense Ann-II mRNA. Neuritogenesis is also impaired by alpha(2)-plasmin inhibitor, antibodies directed against t-PA and u-PA, and E-aminocaproic acid, a lysine analog that inhibits Plg activation and the binding of Plg to Ann-II. Plasmin generation assays reveal a 2-fold increase in plasmin production on NGF-treated PC-12 cells, which can be blocked by a polyclonal antibody directed against the tail region of Ann-H. From these data, we conclude that Ann-II is transcriptionally up-regulated by NGF and that Ann-II-mediated plasmin generation may play an important role during neurite development in the differentiating PC-12 cell.