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Role of PIEZO Channels in Bladder Function and Dysfunction

PIEZO 通道在膀胱功能和功能障碍中的作用

基本信息

批准号：
10417071
负责人：
Gerard L Apodaca
金额：
$ 59.37万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10417071
关键词：
Affect Afferent Neurons Biological Assay Bladder Bladder Dysfunction Bowman&apos s space Cations Cell membrane Cells Communication Cyclophosphamide Cystitis Defect Epithelial Esthesia Event Exocytosis Functional disorder Goals Guns Hyperreflexia Image Ion Transport Knockout Mice Lead Lower urinary tract Mechanics Mediator of activation protein Membrane Membrane Protein Traffic Molecular Conformation Monitor Nature Neuraxis Overactive Bladder Pathway interactions Pelvis Physiological Piezo ion channels Play Process Reflex action Role Sensory Signal Transduction Stimulus Stratified Epithelium Stratum Basale Stretching Surface Testing Time Tissues Urodynamics Urothelial Cell Urothelium Viral afferent nerve allodynia conditional knockout gain of function gain of function mutation insight loss of function mechanical stimulus mechanotransduction multidisciplinary mutant novel patch clamp response tool

项目摘要

Abstract: During filling and voiding, the degree of tension in the bladder wall must be sensed and then relayed to the central nervous system, otherwise bladder dysfunction ensues. While there is a general understanding of the role of sensory neurons (i.e., wall mechanoceptors) in these events, we have fewer insights into how non-neuronal tissues contribute to tension sensation and transduction in the bladder. Our studies focus on the urothelium, which forms the direct interface between the bladder wall and the urinary space. This tissue responds to changes in tension by modulating its ion transport, membrane traffic, and release of mediators, which are hypothesized to alter bladder function, in part via a local urothelial:afferent reflex. However, we still have limited insights into how tension in the plasma membrane of urothelial cells is sensed, what is the nature of the downstream pathways that are activated in response to stretch, or how do these events contribute to bladder function and dysfunction. We hypothesize that urothelial-expressed PIEZO channels act as mechanosensors that in response to bladder filling promote Ca2+ entry, mediator release, and signaling to afferent nerve processes, promoting normal bladder function. We further hypothesize that dysregulation of PIEZO-dependent mechanotransduction in the urothelium will lead to bladder dysfunction. In Specific Aim 1, we will determine if PIEZO channels act as bona fide mechanosensors by demonstrating the following: (i) that functional PIEZO channels are expressed at the surface of urothelial cells; (ii) that urothelial expressed PIEZO channels respond to physiologically relevant stimuli (i.e., bladder filling); (iii) that PIEZO channels are required for mechanically regulated events including membrane traffic and mediator release; (iv) and that expression of loss-of-function or gain-of-function PIEZO mutants will lead to altered urothelial responses. In Specific Aim 2, we seek to understand how PIEZO channels promote urothelial:afferent signaling. Using novel tools, including ex vivo bladder imaging, we will explore the mechanisms by which PIEZO channels stimulate increases in intracellular Ca2+ ([Ca2+]i). Because PIEZO channels rapidly inactivate, there is likely to be a mechanism to amplify the original signal. Thus, we will define whether Ca2+-induced Ca2+ release or membrane depolarization act downstream of PIEZO channels to increase [Ca2+]i. We will also determine if the PIEZO-triggered rise in [Ca2+]i is critical for stimulating exocytosis and mediator release in the urothelium, and if urothelial PIEZO channels modulate urothelial:afferent signaling. In Specific Aim 3, we will begin to explore whether urothelial PIEZO channels contribute to bladder function and/or dysfunction. Specifically, we will determine whether loss of PIEZO expression or function results in bladder underactivity and if PIEZO channels with gain-of-function mutations lead to bladder overactivity. Finally, we will determine whether urothelial-expressed PIEZO channels contribute to the bladder hyperreflexia and pelvic allodynia associated with cyclophosphamide-induced cystitis.

摘要：在膀胱充盈和排尿过程中，必须检测膀胱壁的张力程度