The Influence of Nicotine Withdrawal on Oxygen Sensing

项目总结/摘要 候选人：沃尔曼博士的职业目标是建立一个独立的研究项目， 尼古丁戒断对氧感的影响。研究和职业发展计划已经 精心设计，以提供必要的技能，博士沃尔曼建立一个新颖的，独立的线， 研究，包括以下目标：1）获得进一步的专业知识，在研究方法和 技术，2）发展教学和指导技能，3）发展赠款写作技能和进一步的学术 领导能力环境：主要导师Fregosi博士是研究呼吸的专家， 并将在K99阶段为Aim 1实验提供技术指导。导师博士 Prabhakar是颈动脉窦神经记录和颈动脉体球细胞膜片钳的专家， 博士Oweiss是体内gCaMP成像的专家，并将在研究期间为Wollman博士提供广泛的培训。 K99阶段。两位导师，沿着其他聚集在一起的咨询委员会，将有助于确保博士。 沃尔曼在这个提案上的成功。除了在芝加哥接受Prabhakar医生的实际操作训练 和佛罗里达的Oweiss，这里提出的所有工作都将在Fregosi实验室进行。弗雷戈西博士 NIH的资金和所需的资源，以支持博士.沃尔曼在培训阶段的研究项目。 亚利桑那大学医学院是本提案指导阶段的理想场所， 优秀的教师导师和可用的设施。研究：我的首要假设是，尼古丁 戒断会改变颈动脉体和孤束核（NTS）中的神经元，NTS是 参与了氧气感应。初步数据显示，长期尼古丁暴露对 对缺氧的反应迟钝，然而，大鼠在尼古丁48小时后对缺氧的反应迟钝 戒断为了建立在这些新的初步数据，我设计了以下具体目标。目标1： 测试尼古丁戒断对10%、12%或15%缺氧的缓解反应的影响，并测试尼古丁戒断对缺氧的缓解反应的能力。 来挽救这种反应。目的2：测试尼古丁戒断对氧的影响 颈动脉体的感觉，NTS神经元激活和膈神经的呼吸运动输出， vivo.目的3：探讨尼古丁戒断对颈动脉体烟碱型乙酰胆碱受体功能的影响 血管球细胞和孤束核神经元。这项工作将使用体积描记法，一种独特的 使用纤维光度测定法的体内钙成像和并行膈神经记录的组合，以及 在离体颈动脉体中的血管球细胞和脑干切片中的NTS神经元的全细胞膜片钳。的 拟议的培训计划将使沃尔曼博士学习新的实验技术，成功地完成 拟议的研究和过渡到一个独立的，校外资助的终身职位在顶级（R1） 研究机构。

PROJECT SUMMARY/ABSTRACT CANDIDATE: Dr. Wollman’s career objective is to establish an independent research program investigating the influence of nicotine withdrawal on oxygen sensing. The research and career development plans have been carefully designed to provide the necessary skills for Dr. Wollman to establish a novel, independent line of research and include the following objectives: 1) to gain further expertise in research methodology and techniques, 2) develop teaching and mentoring skills, and 3) to develop grant writing skills and further academic leadership skills. ENVIRONMENT: Primary mentor Dr. Fregosi is an expert in studying breathing both in vivo and in vitro and will provide technical guidance for Aim 1 experiments during the K99 phase. Mentors Dr. Prabhakar is an expert in carotid sinus nerve recordings and patch clamp from carotid body glomus cells, and Dr. Oweiss is an expert in vivo gCaMP imaging, and will provide extensive training to Dr. Wollman during the K99 phase. Both mentors, along with the rest of the assembled advisory committee, will help to ensure Dr. Wollman’s success with this proposal. With the exception of hands on training with Drs. Prabhakar in Chicago and Oweiss in Florida, all of the work proposed here will be carried out in the Fregosi laboratory. Dr. Fregosi has NIH funding and the resources needed to support Dr. Wollman during the training phase of the research project. The University of Arizona College of Medicine is an ideal setting for the mentored phase of this proposal due to the outstanding faculty mentors and facilities available. RESEARCH: My overarching hypothesis is that nicotine withdrawal alters the carotid body and neurons in the nucleus tractus solitarius (NTS), key neural structures that are involved in oxygen sensing. Preliminary data shows that chronic nicotine exposure has no effect on the ventilatory response to hypoxia, however rats show a blunted response to hypoxia after 48 hours of nicotine withdrawal. To build on these novel preliminary data, I have designed the following specific aims. Aim 1: To test the effects of nicotine withdrawal on the ventilatory response to 10, 12, or 15% hypoxia, and to test the ability for an acute dose of nicotine to rescue this response. Aim 2: To test the effects of nicotine withdrawal on oxygen sensing in the carotid body, NTS neuron activation and the respiratory motor output from the phrenic nerve in vivo. Aim 3: To test the effects of nicotine withdrawal on nicotinic acetylcholine receptor function in carotid body glomus cells and NTS neurons in vitro. This work will be carried out using plethysmography, a unique combination of in vivo calcium imaging using fiber photometry, and concurrent phrenic nerve recordings, and whole cell patch clamp of glomus cells in excised carotid bodies and NTS neurons in a brainstem slice. The proposed training plan will allow Dr. Wollman to learn new experimental techniques, successfully complete the proposed research and transition to an independent, extramurally-funded tenure-track position at a top-tier (R1) research institution.