Dissecting the Interoception Circuit that Controls Airway Constriction

项目摘要/摘要 肺是人体内最大的感觉器官之一。肺部长期暴露在过敏原或其他刺激物中已被证明会影响压力、焦虑、抑郁和痴呆症。另一方面，包括冥想中的深呼吸或启功在内的调节呼吸的练习与改善中枢神经系统(CNS)健康有关。然而，脑和肺在分子和细胞水平上的功能性神经解剖学联系仍然很大程度上是未知的。这个R01应用程序建议使用小鼠模型检验假设，即由肺中的内部感受器检测到的过敏原诱导的信号通过迷走神经传入神经元传递到孤束核(NTS)(目标1)，然后从NTS传递到大脑中的其他中央整合体，可能包括下丘脑室旁核(PVH)(目标2)，最后从迷走神经背侧运动核(DMV)传递到下行脊髓传出，再投射回肺(目标3)以感知和调节呼吸道高反应性。这些目标将包括尖端细胞类型特定电路和病毒跟踪技术、LightSheet 3D成像以及候选的多重RNA原位和无偏SNRNAseq方法，以绘制神经电路图并识别外周神经节、NTS和其他大脑区域中信号激活神经元的特征。此外，他们将结合化学遗传学、光遗传学和基于毒素的方法与神经活动读数(包括CFO、体内钙成像、电生理学和肺生理学)相结合，以评估关键的肺、迷走神经、脊髓和大脑连接的功能影响的得失。

PROJECT SUMMARY/ABSTRACT Lung is one of the largest internal sensory organs. Chronic exposure of the lung to allergens or other irritants has been shown to influence stress, anxiety, depression, and dementia. On the other hand, practices that modulate respiration including deep breathing in meditation or Qi Gong are linked to improved central nervous system (CNS) health. However, the functional neuroanatomical connections between brain and lung at the molecular and cellular level remain largely unknown. This R01 application proposes to test the hypothesis, using mouse models, that allergen-induced signals detected by interoceptors in the lung are transmitted via vagal afferent neurons to nucleus tractus solitarius (NTS) (aim 1), and then from NTS to other central integrators in the brain, potentially including paraventricular nucleus (PVH) in the hypothalamus (aim 2), and lastly from dorsal motor nucleus of the vagus (DMV) to descending spinal cord efferents that project back to the lung (aim 3) to sense and regulate airway hyperresponsiveness. The aims will incorporate cutting-edge cell-type specific circuit and viral tracing techniques, lightsheet 3-D imaging, and candidate multiplex RNA in situ and unbiased snRNAseq approaches to map the neural circuits and identify the signature of signal-activated neurons in peripheral ganglia, NTS and other brain regions. In addition, they will combine chemogenetic, optogenetic and toxin-based approaches with neural activity readouts including cFOS, in vivo calcium imaging, electrophysiology, and lung physiology to assess gain and loss of functional effects of key lung, vagal, spinal and brain connections.