Derivation of GABAergic interneurons from mouse embryonic stem cells

从小鼠胚胎干细胞中衍生出 GABA 能中间神经元

• 批准号: 7676876
• 负责人: Asif Mirza Maroof
• 金额: $ 2.83万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-20 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7676876
• 关键词: Adult; Albumins; Autistic Disorder; Brain; Cell Line; Cell Separation; Cell Therapy; Cell Transplantation; Cell Transplants; Cells; Cerebral cortex; Chronic; Derivation procedure; Destinations; Development; Disease; Disease model; ES Cell Line; Embryo; Engineering; Environment; Epilepsy; Erinaceidae; Fibroblast Growth Factor 8; Generations; Goals; Green Fluorescent Proteins; Injection of therapeutic agent; Interneurons; Learning; Medial; Memory; Mental disorders; Methods; Mitotic; Modification; Morphology; Movement; Mus; Neocortex; Neonatal; Neurologic; Neurons; Nodal; Pan Genus; Partial Epilepsies; Pattern; Perception; Pharmaceutical Preparations; Play; Population; Production; Promoter Regions; Protocols documentation; Publishing; Relative (related person); Research; Resistance; Rodent; Role; Schizophrenia; Site; Somatostatin; Source; Staging; Structure; Subgroup; System; Techniques; Telencephalon; Testing; Tetracyclines; Therapeutic Agents; Transactivation; Transgenic Organisms; Transplantation; base; embryonic stem cell; gamma-Aminobutyric Acid; inhibitor/antagonist; migration; morphogens; nerve stem cell; neurochemistry; neuropsychiatry; neurotransmission; postnatal; progenitor; promoter; repaired; research study; stem; tool; transcription factor

DESCRIPTION (provided by applicant): By providing the major source of inhibitory circuitry in the cerebral cortex, GABAergic neurons are required for the synchronous activity necessary to generate sustained oscillations among neurons within a network that are essential during perception, coordinated movement, learning and memory. Cortical deficits in inhibitory neuronal transmission have been implicated in neuropsychiatric disorders such as epilepsy, autism, and schizophrenia. Generating a limitless supply of GABAergic neurons from would allow for the extensive use of these cells in a variety of disease models in order to evaluate their potential for use in cell based therapies. The goal of this proposal is to generate cortical GABAergic interneurons from mouse embryonic stem (ES) cells. Using modifications of published protocols, I will first optimize the production of telencephalic (FoxG1+) progenitors that co-express Nkx2.1, a transcription factor that is required for the specification of major subgroups of cortical interneurons in rodents. Since Nkx2.1 also gives rise to other populations of subcortically generated cells, I have generated an ES cell line that expresses green fluorescent protein under control of the promoter region of Lhx6, a transcription factor whose expression is enriched within Nkx2.1-lineage interneuron precursors and maintained in most of these cells through postnatal development. Using this and other tools, I will direct ES cells towards putative interneuron progenitor (mitotic) or precursor (postmitotic, but not yet migrated into cortex) states and test their capacity to migrate, survive, and differentiate within postnatal mouse cortex. These experiments could have important implications for cell based therapies for medication resistant chronic focal epilepsy. In addition, successful derivation of Nkx2.1-lineage interneuron's from ES cells would provide an invaluable system for the study of intrinsic and extrinsic factors regulating the development of these critical neurons.

描述（由申请人提供）：通过提供脑皮质中的抑制性电路的主要来源，GABA能神经元是必需的同步活动所必需的，即可在网络中产生持续的神经元持续振荡，这在感知，辅助运动，学习，学习和记忆中至关重要。抑制性神经元传播的皮质缺陷与癫痫，自闭症和精神分裂症等神经精神疾病有关。从各种疾病模型中产生无限的GABA能神经元的供应将允许这些细胞广泛使用这些细胞，以评估其在基于细胞疗法中使用的潜力。该建议的目的是从小鼠胚胎（ES）细胞中产生皮质GABA能中间神经元。使用公开协议的修改，我将首先优化远程脑（FOXG1+）祖细胞的产生，该祖细胞是共表达NKX2.1，这是啮齿动物中皮质中间神经元主要亚组规范所需的转录因子。由于NKX2.1还引起了其他下层生成的细胞群体，因此我产生了ES细胞系，该细胞系在控制LHX6的启动子区域的控制下表达绿色荧光蛋白，LHX6的启动子区域（一种转录因子，其表达在NKX2.1-LineAge Intreneuron前体中富集并通过大多数这些细胞在这些细胞中通过肾上腺后发育维持。使用此工具和其他工具，我将引导ES细胞朝推定的中间神经元祖细胞（有丝分裂）或前体（有丝分裂后，但尚未迁移到皮层状态），并测试其在产后小鼠皮层内迁移，生存和区分的能力。这些实验可能对基于细胞的耐药性耐药性慢性局灶性癫痫具有重要意义。此外，从ES细胞中成功推导了NKX2.1-Linege Interneuron的中间神经元，将为研究这些关键神经元发展的内在和外在因素的研究提供宝贵的系统。