Molecular signature of parafacial expiratory neurons

面旁呼气神经元的分子特征

• 批准号: 10750185
• 负责人: DANIEL K MULKEY
• 金额: $ 44.28万
• 依托单位: UNIVERSITY OF CONNECTICUT STORRS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10750185
• 关键词: Abdomen; Address; Adolescent; Air; Alveolar; Anatomy; Attenuated; BHLH Protein; Back; Breathing; Carbon Dioxide; Cell Nucleus; Cells; Characteristics; Chemoreceptors; Development; Differentiated Gene; Disease; Elements; Evaluation; Exhalation; Gene Expression; Gene Expression Profiling; Genes; Genetic; Genetic Markers; Glutamates; Harvest; Hypoxia; Knowledge; Label; Lateral; Lead; Location; Lung; Manuals; Medial; Mediating; Metabolic; Metabolic stress; Molecular; Molecular Profiling; Motor; Motor Neurons; Mus; Muscle; NBPhox protein; Neonatal; Neurons; Phase; Phenotype; Physiological; Pitt-Hopkins syndrome; Population; Positioning Attribute; Process; Pulmonary Ventilation; Research; Respiratory Center; Respiratory System; Rest; Testing; Transcript; Work; autism spectrum disorder; biomarker identification; cell type; complement C2a; experimental study; expiration; human disease; insight; molecular marker; mouse model; neural; recruit; respiratory; response; single-cell RNA sequencing; transcription factor; transcriptome; transcriptome sequencing; ventilation

PROJECT SUMMARY A critical feature of the respiratory system is its ability to adapt to increased metabolic demand by recruitment of expiratory muscles to facilitate alveolar ventilation, a process referred to as active expiration. Active expiration is thought to be mediated by a cluster of neurons located in the lateral parafacial region (pFL). However, despite this physiological significance, the identity of expiratory pFL neurons is not known and unique markers of these cells have not been determined. To address this need, proposed experiments will determine the molecular profile of expiratory pFL neurons and identify unique markers of this population that can be used to selectively label and manipulate this population for detailed functional analysis. We will use RNA-seq to perform an unbiased evaluation of transcript expression of neurons from the pFL region and a nearby respiratory center called the retrotrapezoid nucleus (RTN). This analysis will focus on glutamatergic populations that are expected to include expiratory pFL neurons and RTN neurons. We will use the known molecular profile of RTN neurons as a touchstone for identification of unique pFL markers. To correlate gene expression with function, we will characterize baseline activity and CO2/H+ sensitivity of putative expiratory parafacial neurons back-labelled from an expiratory control center, and we will manually harvest these cells for targeted qPCR to identify common and differentiating genes between RTN and pFL populations. Identifying the transcriptome of pFL neurons will represent a major technical advance in the field of respiratory control as it would make possible the development of targeting strategies to selectively manipulate this population. This work will also provide insight into the genetic organization of neurons and circuits that control breathing, and in doing so help understand how these elements are perturbed in disease.

项目摘要 呼吸系统的一个关键特征是其通过募集代谢物来适应增加的代谢需求的能力。 呼气肌以促进肺泡通气，这一过程称为主动呼气。主动呼气 被认为是由位于外侧面旁区（pFL）的神经元簇介导的。但尽管 这种生理意义，呼气pFL神经元的身份是未知的，这些独特的标志物， 细胞尚未确定。为了解决这一需求，拟议的实验将确定分子 呼气pFL神经元的分布，并确定该群体的独特标志物，可用于选择性地 标记和操作该群体以进行详细的功能分析。我们将使用RNA-seq来执行一个 对来自pFL区域和附近呼吸中枢的神经元的转录表达的无偏评价 称为后斜方核（RTN）。这项分析将集中在预期会出现的过敏人群， 包括呼气pFL神经元和RTN神经元。我们将使用RTN神经元的已知分子谱 作为鉴定独特pFL标记物的试金石。为了将基因表达与功能联系起来，我们将 表征假定的呼气面旁神经元的基线活性和CO2/H+敏感性， 呼气控制中心，我们将手动收获这些细胞进行靶向qPCR，以识别常见的和 区分RTN和pFL群体之间的基因。鉴定pFL神经元的转录组将 代表了呼吸控制领域的一项重大技术进步，因为它将使开发 有选择性地操纵这群人的策略。这项工作也将提供深入了解遗传 神经元和控制呼吸的电路的组织，并在这样做有助于了解这些元素如何 被疾病困扰。