Axonal outgrowth after peripheral nervous system injury by cyclin-dependent kinase inhibitor ; INK4 family protein

细胞周期蛋白依赖性激酶抑制剂导致周围神经系统损伤后轴突生长；

• 批准号: 17591567
• 负责人: MURASE Tsuyoshi
• 金额: $ 2.43万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17591567/
• 关键词: cyclin-dependent kinase inhibitor; peripheral nervous sytem; axonal outgrowth

Cip/Kip family proteins (p21, p27, p57), cyclin-dependent kinase inhibitor, are known to inhibit Rho/Rho-kinase pathway, which inhibits neurite outgrowth. On the other hand, there is no report that INK4 family proteins (p15, p16, p18, p19), which is also cyclin-dependent kinase inhibitor, have an important roles to the peripheral nervous system except cell cycle. Therefore we started this project to investigate whether INK4 family proteins have good effects to the peripheral nervous system and contribute to the functional recovery after peripheral nervous system injury. Initially we performed cloning of INK4 family gene. Then we constructed the vectors with myc-tag at the N- or C- terminal. We investigated the intracellular localization of INK4 family proteins with HEK293 cells. All INK4 family proteins were expressed in the nucleus of HEK293 cells. INK4 family proteins were expressed in dorsal root ganglion neurons by Adenovirus with INK4 family gene. All INK4 family proteins were expressed in the nucleus of dorsal root ganglion neurons and did not influence axonal outgrowth, the number of neurite, and the number of neurite branching. p21 and p27 are known to be translocated from nucleus to cytoplasm by phosphorylation of threonine or serine and therefore we constructed the INK4 family vectors modified by phosphorylation of threonine and serin and investigated whether INK4 family protein was translocated from nucleus to cytoplasm by phosphorylation of threonine and serine. We did not observe the phenomenon of translocation from nucleus to cytoplasm and all INK4 family proteins were expressed in the nucleus.

已知Cip/Kip家族蛋白（p21、p27、p57），细胞周期蛋白依赖性激酶抑制剂，抑制Rho/Rho激酶途径，其抑制神经突生长。另一方面，INK 4家族蛋白（p15，p16，p18，p19）也是细胞周期蛋白依赖性激酶抑制剂，除细胞周期外，对周围神经系统的作用尚未见报道。因此，我们启动了本项目，以研究INK 4家族蛋白是否对周围神经系统有良好的作用，并有助于周围神经系统损伤后的功能恢复。我们首先克隆了INK 4家族基因。然后我们构建了在N-或C-末端带有myc标签的载体。我们用HEK 293细胞研究了INK 4家族蛋白的细胞内定位。所有INK 4家族蛋白在HEK 293细胞的细胞核中表达。用携带INK 4家族基因的腺病毒转染大鼠背根神经节细胞，可使其表达INK 4家族蛋白。所有INK 4家族蛋白在背根神经节神经元的核中表达，并且不影响轴突生长、轴突数目和轴突分支数目。已知p21和p27通过苏氨酸或丝氨酸的磷酸化从细胞核易位到细胞质，因此我们构建了通过苏氨酸和丝氨酸的磷酸化修饰的INK 4家族载体，并研究INK 4家族蛋白是否通过苏氨酸和丝氨酸的磷酸化从细胞核易位到细胞质。我们没有观察到从细胞核到细胞质的易位现象，所有INK 4家族蛋白都在细胞核中表达。

项目成果

IL-1b promotes neurite outgrowth by deactivating RhoA via p38 MAPK pathway

IL-1b 通过 p38 MAPK 通路失活 RhoA 促进神经突生长

• 发表时间: 2008
• 期刊: Biochem Biophys Res Commun 365
• 作者: Murakami M;Demizu Y;Ko Temporin
• 通讯作者: Ko Temporin