Evaluation of Dbx1-derived neurons as the core rhythm generators in mammalian res

Dbx1 衍生神经元作为哺乳动物资源中核心节律发生器的评估

• 批准号: 8006548
• 负责人: Maria Cristina De Guzman Picardo
• 金额: $ 2.61万
• 依托单位: COLLEGE OF WILLIAM AND MARY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8006548
• 关键词: Behavior; Brain; Brain Stem; Breathing; Cell Lineage; Cells; Diagnosis; Electrophysiology (science); Engineering; Etiology; Evaluation; Functional disorder; Generations; Genetic; Genetic Recombination; Health; Human; In Vitro; Interdisciplinary Study; Knock-in Mouse; Knowledge; Lasers; Lesion; Life; Mammals; Membrane; Methods; Molecular; Molecular Genetics; Motor; Motor output; Neuraxis; Neurons; Physiological; Population; Preparation; Property; Reporter; Research Project Grants; Respiration; Respiration Disorders; Role; Site; Slice; System; Technology; Tracer; Transgenes; Transgenic Organisms; Whole-Cell Recordings; developmental genetics; mouse model; neural circuit; neurodevelopment; neuromechanism; preBotzinger complex; public health relevance; recombinase; relating to nervous system; respiratory

DESCRIPTION (provided by applicant): Mammalian breathing is a vital behavior whose underlying neural mechanism originates in the brainstem. This project aims to determine the components of the brainstem neural circuits that generate and control respiration in mammals, including humans, and thus has significant implications for human health. A site in the ventral medulla called preBotzinger Complex (preBotC) is essential for breathing. However, the cellular composition of the preBotC, in terms of the genetic and developmental identity of the different cell populations it contains, and their physiological properties, remain largely unknown. A genetically distinct subpopulation of neurons in the preBotC is hypothesized to form the kernel that generates respiratory rhythm. To evaluate the role of these key neurons, a multidisciplinary research approach is employed that combines molecular genetics and electrophysiology. Recently developed technologies that deliver cell lineage markers via genetic methods, such as site-specific recombination and fluorescent tagging, have greatly impacted neural development studies. Transgenic knock-in mouse models engineered with recombinase-fused cell lineage tracers and reporter transgenes are essential components of the research project. Likewise, characterization of these genetically distinct neuronal populations is carried out through electrophysiological recordings using a unique in vitro brainstem slice preparation that contains essential respiratory neural circuits and allows both cellular-level and systems-level recordings of respiratory motor output. Thus, this research project can evaluate the importance of the key population of neurons in breathing with a multilevel approach: molecular, cellular and system-level properties will be analyzed. Specific Aim 1 will assess the rhythmogenic role of these neurons through reversible genetic silencing and irreversible laser lesioning. Specific Aim 2 will evaluate the membrane properties of the key neurons consistent with their rhythmogenic role through whole-cell recordings. This project will elucidate the neural origins of mammalian respiration. The new knowledge obtained in this project will advance our understanding in the diagnosis and treatment of respiratory disorders that result from dysfunctions in the central nervous system, and provide key new knowledge regarding rhythm generation, which is generally applicable to understanding brain function. PUBLIC HEALTH RELEVANCE: Breathing is a human behavior that is essential in maintaining life. This project aims to reveal the cellular composition of brainstem neural circuits that generate and control breathing rhythms, and to characterize the properties of these cells consistent with their role as rhythm generators. The new knowledge acquired will facilitate the diagnosis and treatment of respiratory disorders with a central neural etiology, and elucidate the neural mechanisms that underlie rhythmic motor behaviors in general.

描述（由申请人提供）：哺乳动物呼吸是一种重要的行为，其潜在的神经机制起源于脑干。该项目旨在确定脑干神经回路的组成部分，这些神经回路产生和控制哺乳动物（包括人类）的呼吸，从而对人类健康产生重大影响。延髓腹侧的一个部位称为preBotzinger复合体（preBotC），是呼吸所必需的。然而，preBotC的细胞组成，在它所包含的不同细胞群的遗传和发育特性方面，以及它们的生理特性，在很大程度上仍然是未知的。假设preBotC中遗传上不同的神经元亚群形成产生呼吸节律的内核。为了评估这些关键神经元的作用，采用了结合分子遗传学和电生理学的多学科研究方法。最近开发的通过遗传方法传递细胞谱系标记的技术，如位点特异性重组和荧光标记，极大地影响了神经发育研究。用重组酶融合的细胞谱系示踪剂和报告转基因工程化的转基因敲入小鼠模型是该研究项目的重要组成部分。同样地，这些遗传上不同的神经元群体的表征是通过使用独特的体外脑干切片制备的电生理记录进行的，所述体外脑干切片制备包含基本的呼吸神经回路，并且允许呼吸运动输出的细胞水平和系统水平的记录。因此，该研究项目可以用多层次的方法评估呼吸中关键神经元群体的重要性：将分析分子，细胞和系统水平的特性。具体目标1将通过可逆的遗传沉默和不可逆的激光损伤来评估这些神经元的节律性作用。具体目标2将通过全细胞记录评价关键神经元的膜特性，与其节律发生作用一致。这个项目将阐明哺乳动物呼吸的神经起源。在这个项目中获得的新知识将促进我们对中枢神经系统功能障碍引起的呼吸系统疾病的诊断和治疗的理解，并提供有关节律生成的关键新知识，这通常适用于理解大脑功能。 公共卫生相关性：呼吸是维持生命所必需的人类行为。该项目旨在揭示产生和控制呼吸节律的脑干神经回路的细胞组成，并表征这些细胞的特性与其作为节律发生器的作用一致。所获得的新知识将有助于诊断和治疗呼吸系统疾病与中枢神经病因，并阐明神经机制，在一般的节奏运动行为。