Department of Molecular Neurobiology, Institute for Developmental Research, Aichi Human Service Center, Aichi 480-0392, Japan.
Department of Tumor Pathology, Gifu University Graduate School of Medicine, Gifu 501-1194, Japan.
Neurosci Res. 2018 Mar;128:50-57. doi: 10.1016/j.neures.2017.08.002. Epub 2017 Aug 10.
Phactr1 (Phosphatase and actin regulator 1) is abundantly expressed in the central nervous system and considered to regulate various neuronal processes through the regulation of protein phosphorylation and actin cytoskeletal organization. In this study, we prepared a specific antibody against Phactr1, anti-Phactr1, and carried out biochemical and morphological analyses of Phactr1 with mouse brain tissues. Western blotting analyses revealed that Phactr1 was expressed in a tissue-dependent profile in the young adult mouse and in a developmental stage-dependent manner in the mouse brain. In primary cultured hippocampal neurons, while Phactr1 was diffusely distributed in the nucleus and cytoplasm, it was visualized in axon and dendrites with partial colocalization with synapses. Phactr1 was also detected in the synaptosomal and postsynaptic density fractions in biochemical fractionation. Immunohistochemical analyses clarified that Phactr1 was differentially expressed in cortical neurons during corticogenesis; the protein was frequently accumulated in the nucleus at the embryonic stage while it came to diffusely distribute in the cell body at the prepubertal stage. The obtained results suggest that Phactr1 takes part in neuronal functions regulated in a spatiotemporal manner.
Phactr1(磷酸酶和肌动蛋白调节剂 1)在中枢神经系统中大量表达,被认为通过调节蛋白磷酸化和肌动蛋白细胞骨架组织来调节各种神经元过程。在这项研究中,我们制备了针对 Phactr1 的特异性抗体,即抗-Phactr1,并使用小鼠脑组织进行了生化和形态分析。Western blot 分析显示,Phactr1 在年轻成年小鼠中以组织依赖性方式表达,并在小鼠大脑中以发育阶段依赖性方式表达。在原代培养的海马神经元中,Phactr1 弥散分布于细胞核和细胞质中,与突触的部分共定位显示其存在于轴突和树突中。Phactr1 也在生化分级分离的突触小体和突触后密度部分中被检测到。免疫组织化学分析阐明了 Phactr1 在皮质发生过程中在皮质神经元中差异表达;该蛋白在胚胎期时经常在核内积累,而在青春期前阶段则扩散分布于胞体中。这些结果表明 Phactr1 参与了时空调节的神经元功能。