Genetic Control of Basal Telencephalic Development

描述（由申请人提供）：我假设了解由Nkx2-1和Lhx6转录因子调控的转录网络（增强子、启动子和下游转录单位）将为阐明包括皮质中间神经元在内的哺乳动物mge衍生细胞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隆起，MGE）是胚胎基底端脑的一个前结构域，可产生多种端脑gaba能神经元细胞类型，包括苍白质投射神经元、纹状体中间神经元（纹状体中间神经元）和苍白质（皮质和海马）神经元。这些神经元的功能障碍与几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。该研究的长期目标是建立在先前研究的基础上，阐明调控mge衍生细胞的规范和分化的转录通路。Nkx2-1转录因子（TF）位于该层次的顶端。心室区（VZ） Nkx2-1表达缺失导致Lhx6和Lhx8(7)表达缺失（Sussel et al., 1999; Flandin et al., 2010）。Lhx6促进皮质IN迁移和小白蛋白（PV）和生长抑制素（Sst）亚型的特化（Liodis et al., 2007; Zhao et al., 2008; Neves et al., 2012）， Lhx8促进胆碱能神经元的分化，Lhx6/8共同调节皮质、纹状体和苍白球（GP）神经元的分化（Flandin et al., 2011; Lopes et al., 2012）。在此，我提出了一些实验，旨在通过信息学、生物化学、小鼠遗传学、发育和细胞生物学的结合，帮助破译一些驱动mge衍生细胞发育的转录网络。有四个具体目标：1。鉴定小鼠MGE发育过程中表达的转录因子（tf）。2. 利用染色质免疫沉淀和DNA测序技术（ChIP-Seq）测定小鼠MGE细胞中Nkx2-1和Lhx6的染色体结合位点，鉴定直接调控基因。3. 描述由Lhx6和Nkx2-1调控的启动子和增强子（近端和远端）的一个子集。4. 小鼠MafB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.