Hamada Nanako, Iwamoto Ikuko, Nishikawa Masashi, Nagata Koh-Ichi
Department of Molecular Neurobiology, Institute for Developmental Research, Aichi Developmental Disability Center, Kasugai, Japan.
Department of Neurochemistry, Nagoya University Graduate School of Medicine, Nagoya, Japan.
Dev Neurosci. 2021;43(1):43-52. doi: 10.1159/000515188. Epub 2021 Apr 1.
MED13L (mediator complex subunit 13-like) is a component of the mediator complex, which functions as a regulator for gene transcription. Since gene abnormalities in MED13L are responsible for neurodevelopmental disorders, MED13L is presumed to play an essential role in brain development. In this study, we prepared a specific antibody against MED13L, anti-MED13L, and analyzed its expression profile in mouse tissues with focusing on the central nervous system. In Western blotting, MED13L exhibited a tissue-dependent expression profile in the adult mouse and was expressed in a developmental stage-dependent manner in brain. In immunofluorescence analyses, MED13L was at least partially colocalized with pre- and post-synaptic markers, synaptophysin, and PSD95, in primary cultured hippocampal neurons. Immunohistochemical analyses revealed that MED13L was relatively highly expressed in ventricular zone surface of cerebral cortex, and was also located both in the cytoplasm and nucleus of neurons in the cortical plate at embryonic day 14. Then, MED13L showed diffuse cytoplasmic distribution throughout the cerebral cortex at the postnatal day (P) 30. In addition, MED13L appeared to be localized in cell type- and developmental stage-specific manners in the hippocampus and cerebellum. These results suggest that MED13L is involved in the development of the central nervous system and synaptic function.
MED13L(中介体复合物亚基13样)是中介体复合物的一个组成部分,其作为基因转录的调节因子发挥作用。由于MED13L中的基因异常与神经发育障碍有关,因此推测MED13L在大脑发育中起重要作用。在本研究中,我们制备了一种针对MED13L的特异性抗体,即抗MED13L,并着重于中枢神经系统分析了其在小鼠组织中的表达谱。在蛋白质印迹法中,MED13L在成年小鼠中呈现出组织依赖性表达谱,并且在大脑中以发育阶段依赖性方式表达。在免疫荧光分析中,在原代培养的海马神经元中,MED13L至少部分地与突触前和突触后标记物、突触素和PSD95共定位。免疫组织化学分析显示,MED13L在大脑皮质的脑室区表面相对高表达,并且在胚胎第14天也位于皮质板神经元的细胞质和细胞核中。然后,在出生后第30天(P30),MED13L在整个大脑皮质中呈现弥漫性细胞质分布。此外,MED13L似乎以细胞类型和发育阶段特异性的方式定位于海马体和小脑中。这些结果表明,MED13L参与中枢神经系统的发育和突触功能。