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

项目摘要/摘要 化学接收是大脑调节呼吸以应对组织变化的机制 CO2/H+。这一过程对于将大脑中的CO2/H+水平保持在有利于 神经功能正常。斜方体后核(RTN)中的化学敏感神经元直接负责 用于增加呼吸活动并将兴奋性投射发送到呼吸节律生成中心 对CO2/H+的反应。延髓核内对化学敏感的星形胶质细胞直接释放对CO2/H+有反应的ATP 激活神经元以调节呼吸活动并通过激活间接收缩RTN小动脉 P2Y2/4受体。小动脉血管收缩防止刺激消失，从而增加神经元和星形细胞 对CO2/H+的响应。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 支持区域功能的血管系统。这项研究的长期目标将是在分子水平上 脑干化学敏感区和非化学敏感区异质性小动脉细胞类型的特征 为呼吸相关疾病的治疗行动提供潜在靶点的领域。这将通过以下方式实现 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。随后的流式细胞术和使用qPCR的嘌呤能受体表达谱将 展示了不同脑区小动脉的独特特性，就像之前在体外看到的那样。通过使用 转基因敲除小鼠模型，小动脉上特异性嘌呤能受体的细胞类型可以描述 对正常呼吸表型的功能性和重要性。这将通过选择性地实现 使用体内和体外研究的嘌呤能受体亚型敲除小鼠模型。体外小动脉切片 记录比较和对比小动脉的行为。全动物体积描记术和病毒击倒 模型证实了体外小动脉记录，并为血管功能障碍的影响提供了证据 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种血管形成，以及提供 潜在治疗疾病状态下呼吸表型的分子靶点。

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.