1Department of Biochemistry and Biophysics, Graduate School of Health Care Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bldg 3, Bunkyo-ku, Tokyo, Japan, 113-8510.
FASEB J. 2014 Mar;28(3):1386-97. doi: 10.1096/fj.13-241034. Epub 2013 Dec 16.
Teneurin-4 (Ten-4), a transmembrane protein, is highly expressed in the central nervous system; however, its cellular and molecular function in neuronal differentiation remains unknown. In this study, we aimed to elucidate the function of Ten-4 in neurite outgrowth. Ten-4 expression was induced during neurite outgrowth of the neuroblastoma cell line Neuro-2a. Ten-4 protein was localized at the neurite growth cones. Knockdown of Ten-4 expression in Neuro-2a cells decreased the formation of the filopodia-like protrusions and the length of individual neurites. Conversely, overexpression of Ten-4 promoted filopodia-like protrusion formation. In addition, knockdown and overexpression of Ten-4 reduced and elevated the activation of focal adhesion kinase (FAK) and Rho-family small GTPases, Cdc42 and Rac1, key molecules for the membranous protrusion formation downstream of FAK, respectively. Inhibition of the activation of FAK and neural Wiskott-Aldrich syndrome protein (N-WASP), which is a downstream regulator of FAK and Cdc42, blocked protrusion formation by Ten-4 overexpression. Further, Ten-4 colocalized with phosphorylated FAK in the filopodia-like protrusion regions. Together, our findings show that Ten-4 is a novel positive regulator of cellular protrusion formation and neurite outgrowth through the FAK signaling pathway.
神经钙黏蛋白-4(Ten-4)是一种跨膜蛋白,在中枢神经系统中高度表达;然而,其在神经元分化中的细胞和分子功能尚不清楚。在本研究中,我们旨在阐明 Ten-4 在轴突生长中的作用。Ten-4 表达在神经母细胞瘤细胞系 Neuro-2a 的轴突生长过程中被诱导。Ten-4 蛋白定位于轴突生长锥。Neuro-2a 细胞中 Ten-4 表达的敲低减少了类丝状伪足的形成和单个轴突的长度。相反,Ten-4 的过表达促进了类丝状伪足的形成。此外,Ten-4 的敲低和过表达分别降低和升高了粘着斑激酶(FAK)和 Rho 家族小 GTP 酶(Cdc42 和 Rac1)的激活,这些是 FAK 下游膜状突起形成的关键分子。FAK 和神经 Wiskott-Aldrich 综合征蛋白(N-WASP)的激活抑制剂,FAK 和 Cdc42 的下游调节剂,阻止了 Ten-4 过表达引起的突起形成。此外,Ten-4 在丝状伪足区域与磷酸化的 FAK 共定位。总之,我们的研究结果表明,Ten-4 通过 FAK 信号通路成为细胞突起形成和轴突生长的新的正调控因子。