Genetic Control of Basal Telencephalic Development

描述（由申请人提供）：我假设理解由Nkx 2 -1和Lhx 6转录因子调控的转录网络（增强子、启动子和下游转录单位）将为阐明哺乳动物MGE衍生细胞（包括皮质中间神经元）发育和功能的转录回路提供关键信息。内侧神经节隆起（medial ganglionic eminence，MGE）是胚胎端脑基底部的一个祖细胞结构域，产生多种端脑GABA能神经元类型，包括苍白球投射神经元、纹状体中间神经元（striatal interneurons，IN）和苍白球（皮层和海马）IN。这些神经元的功能障碍与多种类型的人类神经精神疾病有关，包括癫痫、智力残疾、自闭症和精神分裂症。拟议研究的长期目标是建立在以前的研究基础上，阐明调节MGE衍生细胞的特异性和分化的转录回路。Nkx 2 -1转录因子（TF）位于该层次的顶部。心室区（VZ）中Nkx 2 -1表达的缺失导致Lhx 6和Lhx 8（7）不表达（Sussel等，1999; Festival等人，2010年）。Lhx 6促进皮质IN迁移和小清蛋白（PV）和生长抑素（Sst）亚型的特化（Liodis等人，2007; Zhao等人，2008; Neves等人，2012），Lhx 8促进胆碱能神经元的分化，并且Lhx 6/8一起调节皮质和纹状体IN和苍白球（GP）神经元的分化（Festival等人，2011; Lopes等人，2012年）。在此，我提出了一些实验，旨在有助于破译一些转录网络，驱动MGE衍生细胞的发展，使用信息学，生物化学，小鼠遗传学和发育和细胞生物学的组合。有四个具体目标：1。识别在小鼠MGE发育过程中表达的转录因子（TF）。2.使用染色质免疫沉淀和DNA测序（ChIP-Seq）确定小鼠MGE细胞中Nkx 2 -1和Lhx 6的染色体结合位点，以鉴定直接调控的基因。3.表征由Lhx 6和Nkx 2 -1调控的启动子和增强子（近端和远端）的子集。4. MafB小鼠突变体的分析。

DESCRIPTION (provided by applicant): I hypothesize that understanding the transcriptional networks (enhancers, promoters, and downstream transcription units) regulated by the Nkx2-1 and Lhx6 transcription factors will provide critical information for elucidating the transcriptiona circuits underlying the development and function of mammalian MGE-derived cells including cortical interneurons. The medial ganglionic eminence (MGE), a progenitor domain in the embryonic basal telencephalon, generates several telencephalic GABAergic neuronal cell types, including pallidal projection neurons, striatal interneurons (IN) and pallial (cortical and hippocampal) INs. Dysfunction of these neurons is implicated in several types of human neuropsychiatric disorders, including epilepsy, intellectual disability, autism and schizophrenia. The long-term goal of the proposed research is to build upon previous studies in elucidating the transcriptional circuitry that regulates the specification and differentiation of MGE-derived cells The Nkx2-1 transcription factor (TF) lies at the top of the hierarchy. Loss of Nkx2-1 expression in the ventricular zone (VZ) results in no Lhx6 and Lhx8(7) expression (Sussel et al., 1999; Flandin et al., 2010). Lhx6 promotes cortical IN migration and specification of parvalbumin (PV) and somatostatin (Sst) subtypes (Liodis et al., 2007; Zhao et al., 2008; Neves et al., 2012), Lhx8 promotes differentiation of cholingergic neurons, and Lhx6/8 together regulate differentiation of cortical and striatal INs and globus pallidus (GP) neurons (Flandin et al., 2011; Lopes et al., 2012). Herein, I propose experiments aimed at contributing to deciphering some of the transcriptional networks that drive development of MGE-derived cells using a combination of informatics, biochemistry, mouse genetics and developmental and cell biology. There are four Specific Aims: 1. Identify the transcription factors (TFs) that are expressed during mouse MGE development. 2. Determine the chromosomal binding sites of Nkx2-1 and Lhx6 in mouse MGE cells to identify the directly regulated genes using chromatin immunoprecipitation and DNA sequencing (ChIP-Seq). 3. Characterize a subset of the promoters and enhancers (proximal and distal) regulated by Lhx6 and Nkx2-1. 4. Analysis of the MafB mouse mutant.