Exploring the Role of TRPV4 in Regulating Choroid Plexus Functions and CNS Homeostasis in the Healthy Brain

长远眼光：我的研究计划围绕着识别和表征调节大脑功能的基本分子参与者。概述：中枢神经系统(CNS)浸泡在脉络丛(CP)上皮细胞产生的脑脊液(CSF)中。除了脑脊液的产生外，CP还能主动合成和分泌多种信号分子，影响关键的脑功能，包括神经发生和能量平衡。离子通道激活对CP的分泌功能是必不可少的，因为它们运输合成CP分泌体所需的整合元素。因此，钙离子进入和信号传递是这一机制的关键组成部分，但值得注意的是，这一过程的调节因素在很大程度上仍然不确定。我们检测到CP中存在非选择性的非选择性瞬时受体电位香草酸4型(TRPV4)阳离子通道，并观察到其对CP细胞钙信号的强烈影响。短期目标：我们假设TRPV4是CP功能的主要调节者。通过这三个研究目标，我们将：目标1：建立TRPV4在健康大脑中的存在。利用一种新的转基因报告TRPV4小鼠和免疫组织化学，我们将证实在我们的初步实验中观察到的TRPV4在雄性和雌性动物中的强健表达。目的2：确定TRPV4在正常脑内对CP分泌组的调节作用。在对CP中的TRPV4进行药理学调节后，我们将确定它在健康大脑中钙信号和分泌体组成中的作用。目的3：探讨健康人脑CP功能的增龄性变化。通过使用年轻和老年的健康小鼠，我们将研究衰老对TRPV4和CP功能的影响，以及它对老年脑内维持脑内稳态的影响。HQP培训：拟议的目标考虑了HQP培训的方方面面，包括发展强大的智力、分析和研究技能。提案中概述的实验旨在促进他们对离子通道生物学和脑生理学的兴趣，并将提供分子生物学技术、细胞成像和动物实验方面的高级培训，使他们能够过渡到不同的科学职业。意义：尽管CP对大脑动态平衡的贡献已得到很好的认识，但驱动其复杂功能的分子机制令人惊讶地未被探索。拟议的研究将促进对脉络丛多方面作用的基础知识，并将为中枢神经系统的动态平衡和健康衰老提供新的认识。从更广泛的角度来看，我们的研究洞察力将影响我们对多种脑细胞类型如何相互作用的理解，并对大脑功能的集体调节做出贡献。

Long term vision: My research program centers around the identification and characterization of fundamental molecular participants that regulate brain function. Overview: The central nervous system (CNS) is immersed in the cerebrospinal fluid (CSF) produced by the epithelial cells of the choroid plexus (CP). In addition to the production of CSF, CP also actively synthesize and secrete diverse signaling molecules that influence critical brain functions including neurogenesis and energy homeostasis. Ion channel activation is essential to the secretory functions of CP as they transport the integral elements required for the synthesis of the CP secretome. Calcium entry and signaling is thus, a crucial component of this mechanism but remarkably, the regulators of this process remain largely undetermined. We have detected strong presence of Transient Receptor Potential Vanilloid-type 4 (TRPV4), nonselective, calcium permeant, cation channel in the CP and have observed its ability to robustly influence calcium signaling in the cells of the CP. Short term objectives: We hypothesize that TRPV4 is a master regulator of CP functions. Through the three research aims, we will: Aim 1: Establish the presence of TRPV4 in the healthy brain. Utilizing a novel transgenic reporter TRPV4 mice and immunohistochemistry we will confirm the robust expression of TRPV4 observed in our preliminary experiments, in both male and female animals. Aim 2: Determine the role of TRPV4 in regulating CP secretome in the healthy brain. After pharmacological modulation of TRPV4 in the CP, we will define its role in calcium signaling and secretome composition in the healthy brain. Aim 3: Ascertain the age-related changes in CP functions in the healthy brain. By using young and aged, otherwise healthy mice we will investigate the impact of aging in TRPV4 and CP functions and its influence on the maintenance of brain homeostasis in the aged brain. HQP training: The proposed aims have considered all aspects of HQP training, including the development of strong intellectual, analytical and research skills. The experiments outlined in the proposal are designed to promote their interest in ion-channel biology and brain physiology and will also provide advanced training in molecular biological techniques, cellular imaging and animal experiments, enabling their transition into diverse scientific careers. Significance: Even though the contribution of CP to brain homeostasis is well recognized, the molecular machinery that drives its complex functions are surprisingly unexplored. The proposed studies will advance fundamental knowledge on the multifaceted roles of the choroid plexus and will provide new perceptions on homeostasis and healthy aging in the CNS. On a broader point of view, our investigative insights will impact on our understanding of how multiple brain cell types interact and contribute to the collective regulation of brain functions.