TY - JOUR T1 - Nkx2.1 regulates the proliferation and cell fate of telencephalic astrocytes during embryonic development JF - bioRxiv DO - 10.1101/065169 SP - 065169 AU - Shilpi Minocha AU - Delphine Valloton AU - Yvan Arsenijevic AU - Jean-René Cardinaux AU - Raffaella Dreier Guidi AU - Jean-Pierre Hornung AU - Cécile Lebrand Y1 - 2016/01/01 UR - http://biorxiv.org/content/early/2016/07/21/065169.abstract N2 - The homeodomain transcription factor Nkx2.1 controls cell differentiation of telencephalic GABAergic interneurons and oligodendrocytes. Here, we show that Nkx2.1 additionally regulates astrogliogenesis of the telencephalon from embryonic day (E) 14.5 to E16.5. Our work aims to identify the different mechanisms by which Nkx2.1 controls telencephalic astrogliogenesis. In Nkx2.1-/-, a drastic loss of astrocytes is observed which is not related to cell death. In vivo analysis using BrdU incorporation reveals that Nkx2.1 affects the proliferation of ventral neural stem cells that generate early astrocytes. In vitro neurosphere assays show that Nkx2.1 additionally affects the differentiation step of Nkx2.1-derived astrocytes. Chromatin immunoprecipitation and in vitro co-transfection studies of a Nkx2.1-expressing plasmid indicate that Nkx2.1 binds to the promoter of astroglial differentiation gene GFAP, and regulates its expression. Hence, Nkx2.1 controls astroglial production spatiotemporally in embryos by regulating stem cell division and specification of the contributing Nkx2.1+ precursors.AEPAnterior entopeduncular areaBrdU5-bromo-2’-deoxyuridineCCCorpus callosumCCiCingulate cortexCICingulate bundleCre-ERTtmTamoxifen inducible Cre recombinase fused to the ligand binding domain of the estrogen receptorCspg4Chondroitin sulfate proteoglycan 4 (also known as NG2)DIVDay in vitroEEmbryonic dayEGFPenhanced green fluorescent proteinEYFPenhanced yellow fluorescent proteinGABAergicγ-aminobutyric acidergicGad1Glutamate decarboxylase 1 (also known as GAD67)Gad1-EGFPGad1-EGFP knock-in mouseGFAPGlial fibrillary acidic proteinGLASTGlutamate-aspartate transporterGPGlobus pallidusHICHippocampal commissureIGIndusium griseumIRESInternal ribosome entry siteIZIntermediate zoneKOknockoutLGELateral ganglionic eminenceLHLateral hypothalmusLhx6LIM homeodomain (LIM-hd) geneLPOAlateral preoptic areaLVLateral ventricleMGEMedial ganglionic eminenceMZMarginal zoneMZGMidline zipper gliaNG2Neuron-glial antigen 2Ngn2Neurogenin2Nkx2.1NK2 homeobox 1Nkx2.1-CreMouse with the Cre recombinase under control of the Nkx2.1 promoterOlig2Oligodendrocyte transcription factorpCAGpromoter constructed from following sequences: (C) cytomegalovirus early enhancer element, (A) promoter, the first exon and intron of chicken beta-actin gene, (G) the splice acceptor of the rabbit beta-globin genePOAPreoptic areaRMSRostral migratory streamRosa-EYFPRosa26-lox-STOP-lox-EYFP reporter mouseS100ßSmall EF-hand calcium and zinc binding proteinSEPSeptumSTStriatumSVZSubventricular zoneTSTriangular septal nucleusTUNELTerminal deoxynucleotidyl transferase dUTP nick end labelingVZVentricular zoneWTwild-type ER -