Central Regulation of Sodium Appetite via Synergistic Action of RAAS-sensitive Neurons

项目总结 摄入钠的动机，通常被称为“钠(或盐)胃口”，是一种与生俱来的神经。 由肾素-血管紧张素-醛固酮系统(RAAS)调节的对钠缺乏的反应。受损的 这一系统的功能导致不适当的钠摄取，可能对 心血管健康。K99/R00提案中的学习和职业发展活动旨在 为候选人Jon Resch博士提供必要的培训，使其成为一名独立的调查员 一项研究计划，研究神经控制钠食欲的方法。最近两个RAAS敏感神经元 已有证据表明，群体可以调节钠的食欲：大脑皮质核团中的醛固酮感受神经元 孤束(NTSHsd11b2神经元)和血管紧张素II(AngII)感觉神经元的亚群 穹隆下器(SFO)。重要的是，NTSHsd11b2神经元和Angii敏感的SFO神经元都是必需的 对于缺钠诱导的食欲，NTSHsd11b2神经元需要并发的AngII信号来驱动 快速而强劲的钠消耗。此外，这两个对RAAS敏感的人群都促进钠 通过投射到终纹腹侧床核(VlBNST)摄取。这一点很强烈 提示vlBNST是调节钠摄取的神经过程的关键部位。 然而，vlBNST功能的复杂性和神经化学的异质性构成了显著的 寻找和研究其中调节钠食欲的神经元是一项挑战。为了澄清 控制钠摄取的神经回路，这项拟议的研究将(1)确定Angii的位置 使NTSHsd11b2神经元能够驱动钠摄取的信号，(2)破译RAAS- 敏感的输入到vlBNST，(3)使用高通量的单细胞转录产物来产生一个分子 对vlBNST神经元进行“普查”，以及(4)确定vlBNST“钠摄取”神经元的分子特征。 这些实验的结果将为未来许多关于钠摄取的研究奠定基础。 由BNST和下游回路控制，这些神经元通过这些回路产生动机 摄入钠。拟议的研究和培训将在联合国儿童基金会内分泌科内进行 贝丝以色列女执事医疗中心的医学部，并将确保Resch博士的成功 向科学独立过渡。Resch博士将接受基于CRISPR/CAS9方法的培训 来自他的主要导师布拉德福德·洛厄尔博士的小鼠基因工程和单细胞转录 来自他的顾问委员会成员Evan Rosen博士和Linus Tsai博士。此外，通过收购 上述技术专长、课程安排、出席科学会议和实验室管理 在最初的K99颁奖期间，Resch博士将从他的主要导师那里培养一个独立的 研究钠食欲的神经控制的研究计划。

PROJECT SUMMARY The motivation to consume sodium, often referred to as "sodium (or salt) appetite", is a hard-wired neural response to sodium deficiency regulated by the Renin-Angiotensin-Aldosterone System (RAAS). Impaired function of this system results in inappropriate sodium ingestion that can have deleterious effects on cardiovascular health. The studies and career development activities in this K99/R00 proposal are designed to provide the candidate, Dr. Jon Resch, with the training necessary to become an independent investigator with a research program examining the neural control of sodium appetite. Recently two RAAS-sensitive neuronal populations have been shown to regulate sodium appetite: aldosterone-sensing neurons in the nucleus of the solitary tract (NTSHsd11b2 neurons) and a subpopulation of angiotensin II (AngII)-sensing neurons in the subfornical organ (SFO). Importantly, both NTSHsd11b2 neurons and AngII-sensing SFO neurons are necessary for deficiency-induced sodium appetite, and NTSHsd11b2 neurons require concurrent AngII signaling to drive rapid and robust sodium consumption. Furthermore, both RAAS-sensitive populations promote sodium ingestion via projections to the ventral lateral bed nucleus of the stria terminalis (vlBNST). This strongly suggests that the vlBNST is the critical site where neural processing occurs to coordinate sodium appetite. However, the functional complexity and neurochemical heterogeneity of the vlBNST poses a significant challenge to finding and investigating the neurons within it that regulate sodium appetite. In order to elucidate the neural circuits that control sodium appetite, the proposed research will (1) confirm the site of AngII signaling that enables NTSHsd11b2 neurons to drive sodium appetite, (2) decipher the wiring diagram of RAAS- sensitive inputs to the vlBNST, (3) use high-throughput single-cell transcriptomics to generate a "molecular census" of vlBNST neurons, and (4) identify the molecular signature of vlBNST "sodium appetite" neurons. The results of these experiments will form the foundation for many future studies regarding sodium appetite control by the BNST and the downstream circuits through which these neurons produce the motivation to consume sodium. The proposed research and training will be conducted within the Endocrine Division of the Department of Medicine at Beth Israel Deaconess Medical Center, and will ensure the Dr. Resch's successful transition to scientific independence. Dr. Resch will receive training in CRISPR/Cas9-based methods for mouse genetic engineering from his primary mentor, Dr. Bradford Lowell, and in single-cell transcriptomics from his advisory committee members, Drs. Evan Rosen and Linus Tsai. Furthermore, through acquiring the aforementioned technical expertise, coursework, attendance of scientific meetings, and lab management training from his primary mentor during the initial K99 award period, Dr. Resch will cultivate an independent research program studying the neural control of sodium appetite.