Define Interneuron Subpopulations in the Mouse Spinal Cord during Development

定义发育过程中小鼠脊髓的中间神经元亚群

• 批准号: RGPIN-2016-04880
• 负责人: Zhang, Ying
• 金额: $ 2.04万
• 依托单位: Dalhousie University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=682215
• 关键词: Define; Interneuron; Subpopulations; Mouse; Spinal

Precise organization of neural circuits ensures proper functioning of the central nervous system (CNS). Assembly of these circuits requires correct specification of neuronal populations defined during development. One goal in my research is to understand developmental processes underlying formation of functional groups of spinal neurons. Different progenitor domains in the embryonic spinal cord give rise to the motoneurons (MNs) and different interneuron (IN) classes, each with a unique molecular profile. Distinct types of spinal INs emerge from these progenitor domains, and, as recently revealed, some undergo further subdivision to form heterogeneous populations. As yet, however, little is known about molecular mechanisms that govern such subdivision. *** My lab focuses on an IN group known as V3s as a model system to study subdivision of spinal INs during development. V3 INs arise from a ventral progenitor domain, p3, and express the Sim1 transcription factor at early postmitotic stages. V3 INs are glutamatergic and commissural. In mice, genetic deletion of the entire V3 population prevents animals from exhibiting robust and stable gait. We recently demonstrated that at least two V3 subpopulations, ventral and dorsal, reside in mature spinal cord, each with distinct physiological and anatomical properties. Nevertheless, molecular markers required to define these subpopulations and their function remain unavailable. Moreover, our most recent studies, supported by NSERC funding, suggest that differing physiological properties displayed by V3 subpopulations are present embryonically and that Sim1 may play different roles in regulating their establishment. Therefore, we propose to define molecular markers of V3 subpopulations and mechanisms underlying Sim1 function during development. To do so, we will use transgenic mice to trace V3 INs in wild-type control and Sim1-deficient mice during embryogenesis. We will combine electrophysiology, immunohistochemistry, in situ hybridization and microarray/RNA sequencing screening to conduct the following studies:***Aim I. Define physiological V3 subpopulations at embryonic stages to establish a timeline showing when V3 subpopulations start to display distinctive physiological properties. ****Aim II. Identify specific molecular markers, especially specific transcription factors, of V3 subpopulations to define molecular profiles for different V3 subpopulations. ***Aim III. Investigate molecular pathways that regulate the V3 subpopulation formation by Sim1 to determine which downstream molecular factors are regulated by the Sim1 transcription factor during embryonic development. *** Successful completion of these studies will allow us to distinguish V3 subpopulations and determine their function in spinal circuits. Our work could also pave the way for studies designed to assess formation of other IN subpopulations. **

神经回路的精确组织确保中枢神经系统（CNS）的正常功能。这些回路的组装需要在发育过程中定义的神经元群体的正确规格。 我的研究目标之一是了解脊髓神经元功能群形成的发育过程。胚胎脊髓中不同的祖细胞结构域产生了运动神经元（MN）和不同的中间神经元（IN）类，每个类都具有独特的分子特征。不同类型的脊髓神经元出现从这些祖域，而且，最近发现，一些进行进一步细分，形成异质群体。然而，到目前为止，人们对这种细分的分子机制知之甚少。*** 我的实验室专注于一个称为V3的IN组作为模型系统，以研究发育过程中脊柱IN的细分。V3 IN起源于腹侧祖域p3，并在有丝分裂早期表达Sim 1转录因子。V3 IN是血管性和连合性的。 在小鼠中，整个V3群体的遗传缺失会阻止动物表现出健壮和稳定的步态。我们最近证明，至少有两个V3亚群，腹侧和背侧，居住在成熟的脊髓，每个具有不同的生理和解剖特性。 然而，定义这些亚群及其功能所需的分子标记仍然不可用。此外，我们最近的研究，由NSERC基金支持，表明V3亚群显示不同的生理特性存在于胚胎中，Sim 1可能在调节它们的建立中发挥不同的作用。 因此，我们建议定义V3亚群的分子标记和Sim 1在发育过程中的功能机制。为此，我们将使用转基因小鼠在胚胎发育过程中追踪野生型对照和Sim 1缺陷小鼠中的V3 IN。我们将结合联合收割机电生理学、免疫组化、原位杂交和微阵列/RNA测序筛选技术进行以下研究：* 目的I。在胚胎阶段定义生理V3亚群，以建立显示V3亚群何时开始显示独特生理特性的时间轴。* 目标二。鉴定V3亚群的特异性分子标记，特别是特异性转录因子，以确定不同V3亚群的分子谱。* 目标三。研究通过Sim 1调节V3亚群形成的分子途径，以确定哪些下游分子因子在胚胎发育过程中受到Sim 1转录因子的调节。*** 这些研究的成功完成将使我们能够区分V3亚群，并确定它们在脊髓回路中的功能。我们的工作也可以为旨在评估其他IN亚群形成的研究铺平道路。**