Involvement of Rho-kinase in inflammatory and neuropathic pain through phosphorylation of myristoylated alanine-rich C-kinase substrate (MARCKS)
Involvement of Rho-kinase in inflammatory and neuropathic pain through phosphorylation of myristoylated alanine-rich C-kinase substrate (MARCKS)
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DOI：
10.1016/j.neuroscience.2004.10.022
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发表时间：
2005-01-01
期刊：
影响因子：
3.3
通讯作者：
Ito, S
中科院分区：
文献类型：
--
作者：
Tatsumi, S;Mabuchi, T;Ito, S
富含豆蔻酰丙氨酸的C-激酶底物（MARCKS）是脑中蛋白激酶C的主要体内底物，并且已经涉及与细胞骨架重构相关的细胞过程，例如突触运输和神经递质释放。磷酸化位点特异性抗体pS159-Mar-Ab显示MARCKS在Ser 159处被Rho激酶磷酸化，其磷酸化被Rho激酶特异性抑制剂H-1152抑制。由于MARCKS的功能是由多个位点的磷酸化调节的，在这里，我们研究了Rho激酶与H-1152引起的炎症性和神经性疼痛中MARCKS在Ser 159的磷酸化的关系。当在s.c.注射福尔马林后，10和100 ng H-1152减弱了福尔马林试验中的第二阶段疼痛样行为，但不减弱第一阶段疼痛样行为。鞘内注射H-1152也可缓解选择性L5脊神经切断引起的神经病理性疼痛。一氧化氮合酶活性可视化NADPH黄递酶组织化学福尔马林注射后30分钟和神经切断后7天，这是由H-1152阻断脊髓浅层增加。用pS159-Mar-Ab检测脊髓中MARCKS在Ser 159处的磷酸化，并且在神经切断后的浅层中磷酸化水平增加。与此相反，神经元型一氧化氮合酶和MARCKS的免疫反应性没有显着变化，在脊髓神经切断前后。总之，本研究表明，Rho激酶参与炎症性疼痛和维持神经性疼痛通过磷酸化MARCKS在Ser 159。(C)2005年IBRO。由爱思唯尔有限公司出版。保留所有权利。
Myristoylated alanine-rich C-kinase substrate (MARCKS) is a major in vivo substrate for protein kinase C in the brain and has been implicated in cellular processes associated with cytoskeletal restructuring such as synaptic trafficking and neurotransmitter release. A phosphorylationsite specific antibody against Serl59-phospho-MARCKS (pS159-Mar-Ab) revealed that MARCKS is phosphorylated at Ser159 by Rho-kinase and that its phosphorylation is inhibited by the Rho-kinase specific inhibitor H-1152. Since the function of MARCKS is regulated by phosphorylation at multiple sites, here we examined the involvement of Rho-kinase in relation to phosphorylation of MARCKS at Ser159 in inflammatory and neuropathic pain by H-1152. When intrathecally administered 10 min before s.c. injection of formalin, H-1152 at 10 and 100 ng attenuated the second-phase, but not the first-phase, pain-like behaviors in the formalin test. Neuropathic pain induced by selective L5 spinal nerve transection was also relieved by intrathecal injection of H-1152. Nitric oxide synthase activity visualized by NADPH diaphorase histochemistry increased in the superficial layer of the spinal cord 30 min after formalin injection and 7 days after nerve transection, which were blocked by H-1152. Phosphorylation of MARCKS at Ser159 was detected in the spinal cord by pS159-Mar-Ab and the level of phosphorylation increased in the superficial layer after nerve transection. In contrast, immunoreactivities of neuronal nitric oxide synthase and MARCKS did not change significantly in the spinal cord before and after nerve transection. Taken together, the present study demonstrates that Rho-kinase is involved in inflammatory pain and the maintenance of neuropathic pain through phosphorylation of MARCKS at Ser159. (C) 2005 IBRO. Published by Elsevier Ltd. All rights reserved.