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

探索 TRPV4 在调节健康大脑脉络丛功能和中枢神经系统稳态中的作用

• 批准号: RGPIN-2022-03565
• 负责人: Zechariah, Anil
• 金额: $ 2.04万
• 依托单位: Memorial University of Newfoundland
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=760897
• 关键词: Exploring; Role; TRPV4; Regulating; Choroid

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.

长期愿景：我的研究项目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）浸没在脉络丛（CP）上皮细胞产生的脑脊液（CSF）中。除了CSF的产生，CP还积极合成和分泌影响关键脑功能（包括神经发生和能量稳态）的多种信号分子。离子通道活化对CP的分泌功能是必不可少的，因为它们运输合成CP分泌组所需的整合元素。因此，钙进入和信号传导是这一机制的关键组成部分，但值得注意的是，这一过程的调节剂在很大程度上仍未确定。我们已经检测到CP中的瞬时受体电位香草酸型4（TRPV4）、非选择性、钙渗透性、阳离子通道的强烈存在，并且已经观察到其强烈影响CP细胞中的钙信号传导的能力。短期目标：我们假设TRPV4是CP功能的主要调节因子。通过这三个研究目标，我们将：目标1：确定TRPV4在健康大脑中的存在。利用新型转基因报告基因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稳健表达。目的2：确定TRPV4在正常脑CP分泌组中的调节作用。在CP中TRPV4的药理学调节后，我们将确定其在健康大脑中钙信号传导和分泌组组成中的作用。 目的3：探讨正常人脑CP功能的增龄性变化。通过使用年轻和老年的，否则健康的小鼠，我们将调查的影响TRPV4和CP功能的老化及其对维持大脑稳态的影响在老年大脑。 HQP培训：拟议的目标考虑到了人力资源合格人员培训的所有方面，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，使他们能够过渡到各种科学职业。 重要性：尽管CP对大脑内稳态的贡献是公认的，但驱动其复杂功能的分子机制却令人惊讶地未被探索。拟议的研究将推进脉络丛的多方面作用的基础知识，并将提供新的观念，在中枢神经系统的稳态和健康的老龄化。从更广泛的角度来看，我们的研究见解将影响我们对多种脑细胞类型如何相互作用并有助于集体调节大脑功能的理解。