Role of P2Y2 in the Regulation of Vascular Tone and Control of Breathing

P2Y2 在血管张力调节和呼吸控制中的作用

• 批准号: 9925096
• 负责人: Colin Cleary
• 金额: $ 4.02万
• 依托单位: UNIVERSITY OF CONNECTICUT STORRS
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9925096
• 关键词: Animals; Area; Astrocytes; Award; Behavior; Blood Vessels; Blood flow; Brain; Brain Stem; Brain region; Breathing; Carbon Dioxide; Cell Nucleus; Cell Separation; Cells; Chemicals; Chemoreceptors; Disease; Endothelial Cells; Excision; Expression Profiling; Fellowship; Foundations; Functional disorder; Genotype; Goals; Heterogeneity; In Vitro; Investigation; Knockout Mice; Knowledge; Lead; Letters; Lung diseases; Maintenance; Mediating; Mentors; Mentorship; Metabolic; Methods; Modeling; Molecular; Molecular Target; Neurons; Neurophysiology - biologic function; Nucleus solitarius; P2Y2 receptor; Phase; Phenotype; Physiology; Plethysmography; Population; Postdoctoral Fellow; Process; Property; Protein Isoforms; Purinoceptor; Recording of previous events; Regulation; Reporter; Research; Research Personnel; Research Training; Respiratory physiology; Role; Signal Transduction; Site; Slice; Smooth Muscle; Smooth Muscle Myocytes; Stimulus; Tissues; Transgenic Organisms; Vascular Smooth Muscle; Viral; Whole Body Plethysmography; Work; arteriole; base; cell type; experimental study; in vivo; knock-down; mind control; mouse model; neuroregulation; pre-doctoral; prevent; receptor; receptor expression; relating to nervous system; respiratory; response; targeted treatment; therapeutic target; transcriptome; vasoconstriction

PROJECT SUMMARY/ABSTRACT Chemoreception is the mechanism by which the brain regulates breathing in response to changes in tissue CO2/H+. This process is critical for maintaining brain CO2/H+ levels within the narrow range that is conducive for normal neural function. Chemosensitive neurons in the Retrotrapezoid Nucleus (RTN) are directly responsible for increasing respiratory activity and send excitatory projections to respiratory rhythm generating centers in response to CO2/H+. Chemosensitive astrocytes in the RTN release ATP in response to CO2/H+ that directly gains up neuronal activation to modulate respiratory activity and indirectly constrict RTN arterioles by activation of P2Y2/4 receptors. Arteriole vasoconstriction prevents stimulus washout which gains up neuronal and astrocytic responses to CO2/H+. An unexplored area of brainstem chemoreceptor research is the role heterogenetic vasculature in supporting regional functionality. The long-term goal for this fellowship will be to molecularly characterize heterogenetic arteriole cell types in brainstem chemosensing regions and non-chemosensitive areas to provide potential targets for therapeutic action in breathing related disorders. This will be achieved by using cell isolation methods in transgenic fluorescent mouse models for isolation of both endothelial cells and vascular smooth muscle cells. Subsequent FACS and purinergic receptor expression profiling using qPCR will demonstrate the unique properties of arterioles in different brain regions, as previously seen in vitro. By using transgenic knock out mouse models, specific purinergic receptors on arteriole cell types can profiled for functionality and importance for a normal breathing phenotype. This will be accomplished with selective purinergic receptor isoform knock out mouse models using both in vivo and in vitro studies. In vitro arteriole slice recordings compare and contrast arteriole behavior. Whole animal plethysmography and viral knockdown models confirm in vitro arteriole recordings as well as provide evidence on the impact of vasculature dysfunction in whole animal breathing physiology. These results can be utilized further by other researchers in the field for more in-depth investigation of heterogenetic vasculature in other areas of the brain and body as well as provide molecular targets to potentially remediate breathing phenotypes in disease states.

项目总结/摘要 化学感受是大脑调节呼吸以响应组织变化的机制 CO2/H+。这一过程对于将大脑CO2/H+水平维持在有利于大脑健康的狭窄范围内至关重要。 正常的神经功能梯形后核（RTN）中的化学敏感神经元直接负责 用于增加呼吸活动并向呼吸节律产生中心发送兴奋性投射， 对CO2/H+的反应。RTN中的化学敏感性星形胶质细胞响应于CO2/H+释放ATP， 获得神经元激活以调节呼吸活动并通过激活间接收缩RTN小动脉 P2 Y2/4受体。小动脉血管收缩防止刺激洗脱， 对CO2/H+的反应。脑干化学感受器研究的一个未探索的领域是异质性的作用， 血管系统支持区域功能。这项研究的长期目标是从分子水平上 表征脑干化学敏感区和非化学敏感区中的异质性小动脉细胞类型 为呼吸相关疾病的治疗作用提供潜在靶点的领域。为实现这些目标将 在转基因荧光小鼠模型中使用细胞分离方法分离内皮细胞和 血管平滑肌细胞随后的FACS和使用qPCR的嘌呤能受体表达谱分析将 证明了不同脑区小动脉的独特性质，正如以前在体外观察到的那样。通过使用 在转基因敲除小鼠模型中，可以分析小动脉细胞类型上的特异性嘌呤能受体， 对于正常呼吸表型的功能性和重要性。这将通过选择性的 嘌呤能受体同种型敲除小鼠模型使用体内和体外研究。离体小动脉切片 记录比较和对比小动脉行为。全动物体积描记和病毒敲除 模型证实了体外小动脉记录，并提供了有关血管功能障碍影响的证据 在整个动物呼吸生理学中。这些结果可以被该领域的其他研究人员进一步利用， 更深入地研究大脑和身体其他区域的异质血管系统， 分子靶点以潜在地补救疾病状态中的呼吸表型。