Defining Astrocyte Heterogenity Across the Brainstem Respiratory Circuit

定义脑干呼吸回路中星形胶质细胞的异质性

• 批准号: 9896328
• 负责人: DANIEL K MULKEY
• 金额: $ 24.15万
• 依托单位: UNIVERSITY OF CONNECTICUT STORRS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-18 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9896328
• 关键词: Animal Behavior; Astrocytes; Behavior; Biological Models; Brain; Brain Stem; Brain region; Carbon Dioxide; Cell Nucleus; Cells; Cellular Structures; Central Nervous System Diseases; Chemoreceptors; Complement; Complex; Development; Disease; Electrophysiology (science); Elements; Evaluation; Exhibits; Exposure to; Foundations; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Generations; Genes; Genetic Markers; Genetic Transcription; Genetic Variation; Goals; Harvest; Heterogeneity; Imaging Techniques; Individual; Knowledge; Label; Light; Metabolic; Molecular; Molecular Profiling; Motor; Neurons; Nucleus solitarius; Output; Paracrine Communication; Pathologic; Periodicity; Physiological; Population; Positioning Attribute; Research; Respiration Disorders; Respiratory Center; Rett Syndrome; Role; Signaling Molecule; Slice; Specialized Center; Stimulus; System; Testing; Transcript; Variant; Work; base; cell type; genetic profiling; insight; novel; relating to nervous system; respiratory; response; tool; tool development; transcriptome sequencing

PROJECT SUMMARY Cortical astrocytes are genetically diverse; however, the physiological relevance of this is far from understood because contributions of functionally discrete astrocyte subtypes to whole animal behavior is largely unknown. The most compelling evidence supporting the possibility that astrocytes may have specialized physiological roles is at the level of the brainstem where subsets of astrocytes contribute to the CO2/H+-dependent drive to breathe (respiratory chemoreception) and inspiratory rhythm generation. Therefore, we will use the respiratory circuit as a model system to identify unique genetic profiles of functionally defined subsets of astrocytes. We will use RNA-seq to perform an unbiased evaluation of transcript expression in all cell types isolated from chemosensitive regions, the inspiratory rhythm generating region and non-respiratory brainstem and cortical regions. This analysis will focus on cell populations identified as astrocytes based on expression of cell type specific markers. To correlate gene expression with function, we will use electrophysiological or Ca2+ imaging techniques to functionally identify individual astrocytes in slices from each region for subsequent harvest and targeted qPCR for genes expressed by each astrocyte subpopulation. The proposed work will enable development of tools to study contributions of discrete subpopulations of astrocyte to respiratory behavior. Also, since disruption of astrocyte CO2/H+ chemoreception contributes to disordered breathing in certain diseases (e.g. Rett syndrome), the ability to selectively target CO2/H+-sensitive astrocytes would be an attractive strategy for treating disordered breathing associated with this condition.

项目摘要 皮质星形胶质细胞在遗传上多样。但是，这远未被理解 因为功能离散的星形胶质细胞亚型对整个动物行为的贡献在很大程度上是未知的。 最令人信服的证据支持星形胶质细胞可能具有专门生理的可能性 角色处于脑干的水平，星形胶质细胞子集有助于CO2/H+依赖性驱动器 呼吸（呼吸性化学感受）和灵感节奏产生。因此，我们将使用呼吸道 电路作为模型系统，以识别星形胶质细胞功能定义的子集的独特遗传概况。我们 将使用RNA-Seq在所有从中隔离的细胞类型中对转录本表达的无偏评估 化学敏感区域，灵感节奏产生区域以及非呼吸性脑干和皮质 地区。该分析将重点放在基于细胞类型的表达的星形胶质细胞的细胞群体上 特定标记。为了将基因表达与功能相关，我们将使用电生理或CA2+成像 从功能上识别每个区域中切片中单个星形胶质细胞的技术以进行随后的收获和 针对每个星形胶质细胞亚群表达的基因的靶向qPCR。拟议的工作将启用 开发研究星形胶质细胞对呼吸行为的离散亚群的贡献的工具。 同样，由于星形胶质细胞CO2/H+化学感受的破坏会导致呼吸无序 疾病（例如RETT综合征），选择性靶向CO2/H+敏感的星形胶质细胞的能力将是一种 治疗与这种情况相关的无序呼吸的有吸引力的策略。