Generation of Neuron and Glia Specific Cre Mice

描述(申请人提供)：哺乳动物大脑中神经元的巨大多样性及其散布的排列需要基因靶向来研究在各种神经和精神疾病中已被识别的个体亚群。例如，帕金森氏病中多巴胺能神经元的选择性变性，或GABA能神经元参与脑发育和癫痫。因此，突变特定神经元组或直接或通过神经胶质修饰控制其活动的能力，将是阐明这些神经元以及这些神经元中单个分子对健康和疾病中不同大脑功能的贡献的有力工具。为了帮助对单个亚群的研究，我们计划产生含有基于细菌人工染色体(BAC)的驱动转基因表达他莫昔芬诱导的Cre重组酶(CreERT2)的转基因小鼠，CreERT2是病毒Cre重组酶和Chambon、Metzger及其同事开发的突变的人类雌激素受体配体结合结构域的融合蛋白。胶质细胞将成为GFAP和S100B驱动程序的目标。对于GABA能神经元，我们将使用GAD65(GAD2)和VGAT(Slc32a1)作为驱动器。胺能神经元将以多巴胺转运蛋白(SLC6a3)、多巴胺β-羟基酶(DBH)和色氨酸羟基酶2(TPH2)基因为驱动基因。CreERT2的可诱导性将允许在成熟动物中缺失，并避免在发育过程中因缺失漂浮基因而引起的并发症。随着基因组科学的重心从基因组测序转向功能基因组学，大量带有loxP位点和loxP停止救援的转基因基因的小鼠已经产生。拟议的CreERT2驱动系将提供一个宝贵的工具，从基因上剖析神经系统中的丛生基因的功能，从而有助于识别与疾病相关的基因。与公共卫生相关。我们的建议的医学意义得到了越来越多与疾病相关的基因的发现的支持，特别是在大脑中，可以在密切相关的动物模型中进行研究。我们可以提供转基因动物模型来概括人类大脑疾病，从而促进他们获得新的治疗方法，这将使制药和医学界都受益。

DESCRIPTION (provided by applicant): The enormous diversity of neurons in the mammalian brain and their interspersed arrangement require genetic targeting to study individual subpopulations that have been recognized in various neurological and psychiatric diseases. For example, the selective degeneration of dopaminergic neurons in Parkinson's disease or the involvement of GABAergic neurons in brain development and epilepsies. Therefore, the ability to mutate specific neuronal groups or control their activity directly or via glial modifications, will be a powerful tool for elucidating the contribution of such neurons-and the contribution of individual molecules within these neurons-to diverse brain functions in health and disease. To aid the study of individual subpopulations we plan to generate transgenic mice harboring bacterial artificial chromosome (BAC) based driver-transgenes that express tamoxifen inducible Cre recombinase (CreERT2), which is a fusion protein of the viral Cre recombinase and a human mutated estrogen receptor ligand binding domain developed by Chambon, Metzger and colleagues. Glia cells will be targeted by Gfap and S100b drivers. For GABAergic neurons we will use GAD65 (Gad2) and VGAT (Slc32a1) as drivers. Aminergic neurons will be targeted with the genes for dopamine transporter (Slc6a3), dopamine beta-hydroxylase (Dbh) and tryptophan hydroxylase 2 (Tph2) as drivers. The inducibility of CreERT2 will allow deletion in mature animals and avoid complications that arise from deletion of floxed genes during development. With the focus of genome science shifting from sequencing genomes to functional genomics, numerous mice with genes flanked by loxP sites and loxP stopped rescue transgenes have been generated. The proposed CreERT2 driver lines would provide an invaluable tool to genetically dissect the functions of floxed genes in the nervous system, thus facilitating the identification of disease-linked genes. PUBLIC HEALTH RELEVANCE. The medical relevance of our proposal is supported by the discovery of an increasing number of disease linked genes, especially in the brain, that can be studied in a closely related animal model. We could provide the transgenic animal models to recapitulate human brain disorders, and thereby facilitate their experimental access to new therapeutic approaches, that would benefit both the pharmaceutical and medical communities.