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1420-90717922022Jan30Cellular and molecular life sciences : CMLSCell Mol Life SciGeneration of dystrophin short product-specific tag-insertion mouse: distinct Dp71 glycoprotein complexes at inhibitory postsynapse and glia limitans.10910910910.1007/s00018-022-04151-2Duchenne muscular dystrophy (DMD), the most severe form of dystrophinopathies, is a fatal X-linked recessive neuromuscular disorder characterized by progressive muscle degeneration and various extents of intellectual disabilities. Physiological and pathological roles of the responsible gene, dystrophin, in the brain remain elusive due to the presence of multiple dystrophin products, mainly full-length dystrophin, Dp427, and the short product, Dp71. In this study, we generated a Dp71-specific hemagglutinin (HA) peptide tag-insertion mice to enable specific detection of intrinsic Dp71 expression by anti-HA-tag antibodies. Immunohistochemical detections in the transgenic mice demonstrated Dp71 expression not only at the blood-brain barrier, where astrocytic endfeet surround the microvessels, but also at the inhibitory postsynapse of hippocampal dentate granule neurons. Interestingly, hippocampal cornu ammonis (CA)1 pyramidal neurons were negative for Dp71, although Dp427 detected by anti-dystrophin antibody was clearly present at the inhibitory postsynapse, suggesting cell-type dependent dystrophin expressions. Precise examination using the primary hippocampal culture validated exclusive localization of Dp71 at the inhibitory postsynaptic compartment but not at the excitatory synapse in neurons. We further performed interactome analysis and found that Dp71 formed distinct molecular complexes, i.e. synapse-associated Dp71 interacted with dystroglycan (Dg) and dystrobrevinβ (Dtnb), whereas glia-associated Dp71 did with Dg and dystrobrevinα (Dtna). Thus, our data indicate that Dp71 and its binding partners are relevant to the inhibitory postsynaptic function of hippocampal granule neurons and the novel Dp71-transgenic mouse provides a valuable tool to understand precise physiological expressions and functions of Dp71 and its interaction proteins in vivo and in vitro.© 2022. The Author(s), under exclusive licence to Springer Nature Switzerland AG.FujimotoTakahiroT0000-0002-9975-7008Department of Pathology and Applied Neurobiology, Kyoto Prefectural University of Medicine, Graduate School of Medical Science, 465 Kajii-cho, Kawaramachi Hirokoji, Kamigyo-ku, Kyoto, 602-8566, Japan.YaoiTakeshiTDepartment of Pathology and Applied Neurobiology, Kyoto Prefectural University of Medicine, Graduate School of Medical Science, 465 Kajii-cho, Kawaramachi Hirokoji, Kamigyo-ku, Kyoto, 602-8566, Japan.NakanoKentaKDepartment of Laboratory Animal Medicine, Research Institute, National Center for Global Health and Medicine (NCGM), Tokyo, 162-8655, Japan.AraiTetsuyaTDepartment of Laboratory Animal Medicine, Research Institute, National Center for Global Health and Medicine (NCGM), Tokyo, 162-8655, Japan.OkamuraTadashiTDepartment of Laboratory Animal Medicine, Research Institute, National Center for Global Health and Medicine (NCGM), Tokyo, 162-8655, Japan.ItohKyokoK0000-0001-8369-8800Department of Pathology and Applied Neurobiology, Kyoto Prefectural University of Medicine, Graduate School of Medical Science, 465 Kajii-cho, Kawaramachi Hirokoji, Kamigyo-ku, Kyoto, 602-8566, Japan. kxi14@koto.kpu-m.ac.jp.eng18K07883Japan Society for the Promotion of Science21K07279Japan Society for the Promotion of Science20A1019Grants-in-Aid for Research from the National Center for Global Health and Medicine21A1018Grants-in-Aid for Research from the National Center for Global Health and MedicineJournal Article20220130
SwitzerlandCell Mol Life Sci97054021420-682X0Dtna protein, mouse0Dystrophin0Dystrophin-Associated Proteins0Neuropeptides0apo-dystrophin 1146888-27-9DystroglycansIMAnimalsBlood-Brain BarriermetabolismCells, CulturedDystroglycansgeneticsmetabolismDystrophingeneticsmetabolismDystrophin-Associated ProteinsgeneticsmetabolismHEK293 CellsHippocampuscytologymetabolismHumansMice, TransgenicMicroscopy, ConfocalNeurogliametabolismNeuronsmetabolismNeuropeptidesgeneticsmetabolismProtein BindingSynapsesmetabolismDMDDp71DystrophinGlia limitansInhibitory postsynapseTransgenic mouseThe authors state no conflicts of interest.2021111720221142021122420221316520222160202228602022130epublish35098363PMC1107172510.1007/s00018-022-04151-210.1007/s00018-022-04151-2Deburgrave N, Daoud F, Llense S, Barbot JC, Récan D, Peccate C, Burghes AH, Béroud C, Garcia L, Kaplan JC, Chelly J, Leturcq F. 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