The Influence of Nicotine Withdrawal on Oxygen Sensing

项目摘要/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计划，调查 尼古丁戒断对氧感觉的影响。研究和职业发展计划一直是 精心设计，为沃尔曼博士提供必要的技能，以建立一种新的、独立的 研究和包括以下目标：1)在研究方法和方法方面获得进一步的专门知识 技术，2)发展教学和指导技能，以及3)发展助学金写作技能和进一步的学术 领导才能。环境：主要导师Fregosi博士是研究体内呼吸的专家 并将为目标1号在K99阶段的实验提供技术指导。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方面的专家，以及 奥韦斯博士是活体GCaMP成像方面的专家，他将在 K99相。这两位导师以及咨询委员会的其他成员将帮助确保Dr。 沃尔曼在这项提议上的成功。除了在芝加哥与Prabhakar博士一起动手培训外 和佛罗里达州的奥韦斯，这里提出的所有工作都将在Fregosi实验室进行。Fregosi博士有 NIH的资金和在研究项目的培训阶段支持Wollman博士所需的资源。 亚利桑那大学医学院是该提案指导阶段的理想环境，因为 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 戒断改变颈动脉小体和孤束核(NTS)神经元，这些关键神经结构 都与氧气感应有关。初步数据显示，长期接触尼古丁对 低氧的换气反应，然而，大鼠在尼古丁48小时后对低氧的反应迟钝 戒烟。为了建立在这些新颖的初步数据的基础上，我设计了以下具体目标。目标1：实现 测试尼古丁戒断对10%、12%或15%缺氧的呼吸反应的影响，并测试其能力 急性剂量的尼古丁来挽救这种反应。目的2：检测尼古丁戒断对氧的影响 大鼠颈动脉体区NTS神经元的激活和膈神经的呼吸运动输出 活着。目的：检测尼古丁戒断对颈动脉体乙酰胆碱受体功能的影响 球体细胞和NTS神经元的体外培养。这项工作将使用体积描记术进行，这是一种独特的 结合使用纤维光度法的体内钙成像和同时进行的膈神经记录，以及 全细胞膜片钳技术，对切除的颈动脉小体中的球状细胞和脑干切片中的NTS神经元进行观察。这个 拟议的培训计划将使沃尔曼博士能够学习新的实验技术，成功地完成 拟议的研究和过渡到一个独立的、由外部资金资助的顶级终身教职职位(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.