Biochemical and morphological characterization of SEPT1 in mouse brain

Ito, Hidenori; Morishita, Rika; Noda, Mariko; Iwamoto, Ikuko; Nagata, Koh‐ichi

doi:10.1007/s00795-020-00248-4

Cited by 8 publications

(11 citation statements)

References 25 publications

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“…Cell Culture and Transfection COS7, Neuro-2a (N2a; mouse neuroblastoma) cells, and primary cultured hippocampal neurons derived from the embryonic day 16.5 (E16.5) mice were cultured essentially as described [6]. N2a cell differentiation was induced by serum-starvation for 30 h. To prepare primary cultured cortical and hippocampal neurons, cortices or hippocampi were dissected from E14 mice and collected in Hanks' balance salt solution.…”

Section: Preparation Of Extracts From Mouse Tissuesmentioning

confidence: 99%

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

et al. 2021

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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.

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Section: Preparation Of Extracts From Mouse Tissuesmentioning

confidence: 99%

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

et al. 2021

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“…Sample preparation was performed essentially as described [22][23][24] . Brie y, tissues from ICR adult mice (Japan SLC, Shizuoka, Japan) were homogenized with the lysis buffer (50 mM Tris-HCl, pH 7.5, 0.1% NaF, 5 mM EDTA, 1 mM Na 3 VO 4 and the protease inhibitor mix).…”

Section: Preparation Of Extracts From Mouse Tissues and Western Blottingmentioning

confidence: 99%

“…Protein concentration was estimated with BCA protein assay reagent kit (Pierce, Rockford, IL) with bovine serum albumin as a standard. SDS-PAGE (10 % gel) and western blotting were performed as described [22][23][24] .…”

Section: Preparation Of Extracts From Mouse Tissues and Western Blottingmentioning

confidence: 99%

“…Sample preparation was performed essentially as described 23 . Brie y, adult mouse brains were homogenized in 5 mM HEPES (pH 7.4) containing 1 mM MgCl 2 , 0.5 mM CaCl 2 , phosphatase inhibitors and the protease inhibitor mix with a Te on homogenizer (H fraction).…”

Section: Preparation Of Postsynaptic Densitymentioning

confidence: 99%

“…Cell culture, transfection and immuno uorescence COS7 cell transfection experiments were performed with polyethyleneimine "MAX" reagent (Polysciences Inc., Warrington, PA). Immuno uorescence analyses were carried out as described [22][23][24] . Alexa Fluor 488-, 594-or 647-labeled IgG (Life Technologies) was used as a secondary antibody.…”

Section: Immunohistochemical Analysesmentioning

confidence: 99%

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Physiological Significance of WDR45, A Responsible Gene for β-propeller Protein Associated Neurodegeneration (BPAN), in Brain Development

Noda

Ito

Nagata

2020

Preprint

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WDR45 plays an essential role in the early stage of autophagy. De novo heterozygous mutations in WDR45 have been known to cause b-propeller protein-associated neurodegeneration (BPAN), a subtype of neurodegeneration with brain iron accumulation (NBIA). Although BPAN patients display global developmental delay including intellectual disability, neurodevelopmental pathophysiology of BPAN remains largely unknown. In the present study, we analyzed the physiological role of Wdr45 and pathophysiological significance of the gene abnormality during mouse brain development. Morphological and biochemical analyses revealed that Wdr45 is expressed in a developmental stage-dependent manner in mouse brain. Wdr45 was also found to be located in the excitatory synapses in biochemical fractionation. Since the WDR45 mutations are thought to cause protein degradation, we conducted acute knockdown experiments by an in utero electroporation method with mice to recapitulate the pathophysiological conditions of BPAN. Silencing of Wdr45 caused abnormal dendritic development and synaptogenesis during corticogenesis, both of which were significantly rescued by co-expression with RNAi-resistant version of Wdr45. In addition, terminal arbors of callosal axons were less developed in Wdr45-deficient cortical neurons of adult mouse when compared to the control cells. These results strongly suggest a pathophysiological significance of WDR45 gene abnormalities in neurodevelopmental aspects of BPAN.

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Expression analyses of PLEKHG2, a Rho family-specific guanine nucleotide exchange factor, during mouse brain development

et al. 2021

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Biochemical and morphological characterization of SEPT1 in mouse brain

Cited by 8 publications

References 25 publications

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

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

Physiological Significance of WDR45, A Responsible Gene for β-propeller Protein Associated Neurodegeneration (BPAN), in Brain Development

Expression analyses of PLEKHG2, a Rho family-specific guanine nucleotide exchange factor, during mouse brain development

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