Expression Analyses of Mediator Complex Subunit 13-Like: A Responsible Gene for Neurodevelopmental Disorders during Mouse Brain Development

Abstract

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.

Expression Analyses of Cep152, a Responsible Gene Product for Autosomal Recessive Primary Microcephaly, during Mouse Brain Development

“…Transient transfection was carried out using PEImax reagent according to the manufacturer's instruction (Polysciences, Warrington, PA). Immunoprecipitation was conducted as described [15]. Briefly, COS7 cells expressing Myc-Cep152 were homogenized with 20 mM Hepes/pH 7.6, 50 mM NaCl, 2.5 mM MgCl 2 , 0.1 mM EDTA, 0.5% Triton X-100, 1 mM phenylmethylsulfonyl fluoride, 10 μg/mL aprotinin, and 10 μg/mL leupeptin.…”

Expression Analyses of Rac3, a Rho Family Small GTPase, during Mouse Brain Development

“…Immunohistochemical analysis was carried out essentially as described [14]. Mouse brains of either sex fixed with 4% paraformaldehyde at postnatal day 2 or 18 were cut into coronal or sagittal sections (14 μm thickness) with a cryostat (Leica CM1900, Leica Microsystems, Wetzlar, Germany).…”

Transcription Pause and Escape in Neurodevelopmental Disorders

“…However, the exact mechanism and pathways remain unknown. In a recent mouse study, Hamada et al (2021) showed that MED13L protein is highly expressed in the ventricular zone of the cerebral cortex and is also detectable in the developing hippocampus and cerebellum.…”