Molecular control of neurogenesis in the adult subventricular zone

成人室下区神经发生的分子控制

• 批准号: 7853503
• 负责人: KENNETH J CAMPBELL
• 金额: $ 44.64万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7853503
• 关键词: Address; Adult; Alleles; Animals; Ara-C; Astrocytes; Biological Assay; Biology; Brain; Brain Injuries; Brain Pathology; Bromodeoxyuridine; Cell Culture Techniques; Cell Differentiation process; Cell division; Cells; Corpus striatum structure; Data; Deoxyuridine; Development; Embryo; Gene Expression Profile; Generations; Goals; Growth Factor; Health; Heterogeneity; Human; In Vitro; Injury; Interneurons; Knock-out; Label; Life; Light; Location; Maintenance; Mediating; Mitotic; Molecular; Molecular Genetics; Mus; Natural regeneration; Neurobiology; Neuroglia; Neurology; Neuronal Differentiation; Neurons; Oligodendroglia; Outcome; Outcome Study; Physiological; Physiology; Play; Production; Property; Proteins; Quinolinic Acid; Research; Role; Social Welfare; Source; Specificity; Stream; Testing; Tetanus Helper Peptide; Transgenes; Transgenic Organisms; Travel; Virus; adult neurogenesis; base; chromatin immunoprecipitation; gain of function; genetic analysis; genome-wide; gliogenesis; homeodomain; in vivo; innovation; insight; lateral ventricle; loss of function; molecular marker; nerve stem cell; neuroblast; neurogenesis; neuronal replacement; novel; olfactory bulb; overexpression; progenitor; promoter; public health relevance; repaired; research study; subventricular zone; transcription factor; Address; Adult; Alleles; Animals; Ara-C; Astrocytes; Biological Assay; Biology; Brain; Brain Injuries; Brain Pathology; Bromodeoxyuridine; Cell Culture Techniques; Cell Differentiation process; Cell division; Cells; Corpus striatum structure; Data; Deoxyuridine; Development; Embryo; Gene Expression Profile; Generations; Goals; Growth Factor; Health; Heterogeneity; Human; In Vitro; Injury; Interneurons; Knock-out; Label; Life; Light; Location; Maintenance; Mediating; Mitotic; Molecular; Molecular Genetics; Mus; Natural regeneration; Neurobiology; Neuroglia; Neurology; Neuronal Differentiation; Neurons; Oligodendroglia; Outcome; Outcome Study; Physiological; Physiology; Play; Production; Property; Proteins; Quinolinic Acid; Research; Role; Social Welfare; Source; Specificity; Stream; Testing; Tetanus Helper Peptide; Transgenes; Transgenic Organisms; Travel; Virus; adult neurogenesis; base; chromatin immunoprecipitation; gain of function; genetic analysis; genome-wide; gliogenesis; homeodomain; in vivo; innovation; insight; lateral ventricle; loss of function; molecular marker; nerve stem cell; neuroblast; neurogenesis; neuronal replacement; novel; olfactory bulb; overexpression; progenitor; promoter; public health relevance; repaired; research study; subventricular zone; transcription factor

DESCRIPTION (provided by applicant): Active production of new neurons and glia continues in restricted regions of the adult mammalian brain, including the subventricular zone (SVZ) lining the lateral ventricle. In these so-called neurogenic niches, neural stem cells (NSCs) and other progenitors (collectively called NPCs) persist throughout life and serve as the source of new neurons and glia. Recent studies have revealed that this continuous neurogenesis in adults contribute to physiology and pathology of the brain. Moreover, regenerating specific types of neurons after damage by manipulating endogenous NPCs is one of the important goals in restorative neurology. Yet, our understanding of the mechanisms underlying this important phenomenon is still very limited, and further research is imperative. In this proposal, we will address this issue by studying the roles for the homeodomain transcription factors Gsx1 and Gsx2 in adult neurogenesis. Gsx1 and Gsx2 control many aspects of telencephalic development, including regional specification and proliferation of NPCs, and neuronal differentiation. Their function in adults, however, has not yet been investigated. Our preliminary data show that Gsx1 and Gsx2 are expressed in specific subpopulations of NPCs that reside in anatomically discrete subdomains of the adult SVZ. We hypothesize that Gsx1 and Gsx2 play crucial roles in controlling the maintenance and/or differentiation of discrete subpopulations of NPCs in the adult SVZ, thereby contributing to the generation of specific OB interneuron subtypes. We also hypothesize that Gsx1+ and Gsx2+ progenitors contribute to injury-induced neurogenesis in the adult brain. We will test these hypotheses by experiments outlined in the following four specific aims: Aim 1: To reveal the identity of Gsx1+ and Gsx2+ progenitors in the adult SVZ. Our preliminary data show that Gsx1 and Gsx2 are expressed in NPCs that reside in discrete subdomains of the adult SVZ. We will reveal the properties of these cells by 1) detailed molecular marker analysis using Gsx2flox/+ and Gsx1::GFP mice, 2) short-term and long-term labeling with the mitotic marker 5-bromo-2'deoxyuridine (BrdU) combined with treatment with the anti-mitotic agent b-D-arabinofuranoside (Ara-C) and various growth factors, and 3) clonal cell culture assays. Aim 2: To reveal the roles for Gsx1 and Gsx2 in neurogenesis and gliogenesis in the adult SVZ. To reveal the adult-specific function of Gsx1 and Gsx2, we will perform conditional gain-of-function (GOF) and loss-of-function (LOF) analyses in vivo. Mice carrying floxed alleles of Gsx1 and Gsx2 will be used for progenitor subtype-specific knockout in the adult brain. We will also perform conditional GOF experiments using mice carrying tet-O promoter-driven Gsx1 and Gsx2 transgenes. Virus-mediated overexpression and knockdown of Gsx1 and Gsx2 will be performed in parallel. Aim 3: To reveal the function of Gsx1 and Gsx2 using in vitro culture. To better understand the mechanisms of action of Gsx1 and Gsx2, we will perform GOF and LOF analyses using clonal culture of NPCs. We will also identify the transcriptional targets for Gsx1 and Gsx2 by genome-wide chromatin-immunoprecipitation (ChIP) and transcriptome assays. Aim 4: To reveal the roles for Gsx1 and Gsx2 in injury-induced neurogenesis in the adult brain The mechanisms underlying injury-induced neurogenesis in the adult brain remain largely unknown. We will examine if Gsx1+ and Gsx2+ progenitors participate in ectopic neurogenesis following quinolinic acid (QA)-induced excitotoxic injury in the striatum. We will also determine if Gsx1 and Gsx2 play any role in injury-dependent control of SVZ progenitors and neurogenesis through GOF and LOF analyses using the aforementioned transgenic and virus-mediated approaches. Revealing the roles of Gsx1/2 in the adult brain will provide novel insights into the mechanisms for adult neurogenesis, in particular, the molecular basis for the specificity of adult NPCs. Thus, this study will significantly advance our understanding of adult neurogenesis and the biology of NPCs. Given the importance of adult neurogenesis in physiology and pathology of the brain, the outcomes of this study will have significant contribution to the promotion of human health and welfare. PUBLIC HEALTH RELEVANCE: Continuous production of new neurons in the adult brain (adult neurogenesis) has been shown to play an important role in higher brain functions as well as repair of damaged brains. Yet, our understanding of the mechanisms underlying this important phenomenon is still limited. This study will address this issue by studying the roles of the transcription factors Gsx1 and Gsx2 in adult neurogenesis. Thus, the outcomes of the proposed research will have significant contribution to the promotion of human health and welfare.

描述（由申请人提供）：在成年哺乳动物大脑的受限制区域，包括侧心室侧壁的室内哺乳动物区域（SVZ）的有限区域继续积极产生新的神经元和神经胶质。在这些所谓的神经源性壁ches中，神经干细胞（NSC）和其他祖细胞（共同称为NPC）一生都持续存在，并充当新的神经元和神经胶质的来源。最近的研究表明，成人的这种连续神经发生有助于大脑的生理和病理学。此外，通过操纵内源性NPC损害后，再生特定类型的神经元是恢复性神经病学的重要目标之一。然而，我们对这种重要现象的机制的理解仍然非常有限，并且必须进行进一步的研究。在此提案中，我们将通过研究副域转录因子GSX1和GSX2在成人神经发生中的作用来解决这个问题。 GSX1和GSX2控制了脑胶凝发育的许多方面，包括NPC的区域规范和扩散以及神经元分化。但是，他们在成年人中的功能尚未得到研究。我们的初步数据表明，GSX1和GSX2以NPC的特定亚群来表达，这些NPC位于成年SVZ的解剖学子域中。我们假设GSX1和GSX2在控制成人SVZ中NPC的离散亚群的维持和/或分化中起着至关重要的作用，从而有助于产生特定的OB Interneuron子类型。我们还假设GSX1+和GSX2+祖细胞有助于损伤引起的成人大脑神经发生。 我们将通过以下四个特定目的中概述的实验来检验这些假设：目标1：揭示成人SVZ中GSX1+和GSX2+祖细胞的身份。我们的初步数据表明，GSX1和GSX2在驻留在成人SVZ的离散子域的NPC中。我们将通过1）使用GSX2Flox/+和GSX1 :: GFP小鼠详细揭示这些细胞的特性，2）使用有丝分裂标记物5-溴-2'Deoxyuridine（BRDU）与抗抗溶剂抗合性因子和三核酸素化剂（3-溴）和长期标记（BRDU）和长期标记。克隆细胞培养分析。目标2：揭示GSX1和GSX2在成年SVZ中神经发生和神经胶质发生中的作用。为了揭示GSX1和GSX2的成人特异性功能，我们将在体内执行有条件的功能获得（GOF）和功能丧失（LOF）分析。携带GSX1和GSX2等位基因的小鼠将用于成人大脑的祖细胞亚型特异性敲除。我们还将使用携带Tet-O启动子驱动的GSX1和GSX2转基因的小鼠进行条件GOF实验。病毒介导的GSX1和GSX2的过表达和敲低将平行进行。目标3：使用体外培养物揭示GSX1和GSX2的功能。为了更好地了解GSX1和GSX2的作用机理，我们将使用NPC的克隆培养进行GOF和LOF分析。我们还将通过全基因组染色质免疫沉淀（CHIP）和转录组测定法确定GSX1和GSX2的转录靶标。 AIM 4：揭示GSX1和GSX2在成年大脑中损伤诱导的神经发生中的作用，成年大脑损伤诱导的神经发生的机制仍然很大未知。我们将检查GSX1+和GSX2+祖细胞是否参与了喹啉酸（QA）诱导的纹状体兴奋性损伤后的异位神经发生。我们还将通过上述转基因和病毒介导的方法来确定GSX1和GSX2是否通过GOF和LOF分析在对SVZ祖细胞的损伤依赖性控制和神经发生中起着任何作用。 揭示GSX1/2在成年大脑中的作用将提供有关成人神经发生机制的新见解，特别是成人NPC特异性的分子基础。因此，这项研究将显着提高我们对成年神经发生和NPC生物学的理解。鉴于成年神经发生在大脑的生理和病理学中的重要性，这项研究的结果将对促进人类健康和福利有重大贡献。 公共卫生相关性：已显示成人大脑中新神经元的连续产生（成人神经发生）已被证明在较高的大脑功能以及修复受损的大脑中起着重要作用。然而，我们对这种重要现象的机制的理解仍然有限。这项研究将通过研究转录因子GSX1和GSX2在成人神经发生中的作用来解决这个问题。因此，拟议研究的结果将为促进人类健康和福利做出重大贡献。