Nitric Oxide in Bladder Neural-Epithelial Signaling

膀胱神经上皮信号传导中的一氧化氮

• 批准号: 9119803
• 负责人: LORI A BIRDER
• 金额: $ 30.24万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-15 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9119803
• 关键词: Area; Bladder; Bladder Dysfunction; Bladder Urothelial Cell; Calcium Oscillations; Cell Communication; Cell Surface Receptors; Cell surface; Cells; Chemical Stimulation; Chemicals; Clinical Management; Coculture Techniques; Communication; Coupled; Data; Dinoprostone; Dyes; Epithelial; Epoprostenol; Event; Exhibits; Exocytosis; Fluorescent Dyes; Funding; Future; Goals; Image; Imaging Techniques; Ion Channel; Link; Mechanical Stimulation; Mechanics; Mediator of activation protein; Membrane; Molecular; Movement; Nerve; Nervous system structure; Neurons; Nitric Oxide; Nociception; Pathology; Pathway interactions; Pharmacology; Physical environment; Play; Property; Proteins; RNA Interference; Receptor Cell; Recycling; Research; Role; Sensory; Series; Signal Transduction; Small Interfering RNA; Stimulus; TRP channel; TRPV1 gene; Techniques; Tissues; Toxin; Transducers; Urothelial Cell; Urothelium; Vesicle; cellular targeting; chemical addition; chemical release; detector; imaging modality; insight; intercellular communication; interdisciplinary approach; novel; optical imaging; patch clamp; protein function; relating to nervous system; research study; response; sensor; sensory mechanism; urinary bladder epithelium

DESCRIPTION (Provided by Applicant): Traditionally sensory signaling in the urinary bladder has been attributed to direct activation of bladder afferents. New findings have highlighted urinary bladder urothelial cells as key players in the transduction of sensory events. We have shown that urothelial cells exhibit a number of "neuron-like" properties (including expression of sensor molecules that allow them to respond to chemical/thermal/mechanical stimuli and to release chemical mediators) making it likely that urothelial cells communicate "directly" with bladder nerves or indirectly via urothelial cell-cell interactions. Urothelial cells are constantly exposed to various forms of mechanical stimuli, yet the mechanisms by which these cells respond are not well defined. Mechanical forces are thought to "initiate" a series of signaling events (calcium waves) throughout the urothelium. In turn, the accompanying release of chemical factors induced by mechanical stimuli could activate urothelial cell surface receptors and amplify the signal within and adjacent to the urothelial cell. Although these data suggest that urothelial cells play an important role in cell-cell signaling, this is a relatively unexplored area with little information known about the mechanism for sensory transduction or the mode of communication between cells. Using a multidisciplinary approach involving pharmacology, siRNA, transmitter release, patch clamp and novel imaging techniques, our goals are to evaluate how urothelial cells receive and integrate multiple stimuli. Specific Aim 1 will characterize urothelial cell mechanosensors and their mechano-transduction pathways. Evidence has shown that TRPV1 is essential for mechanically-evoked purinergic signaling by the urothelium, yet the mechanisms by which urothelial cells respond to mechanical forces are not well defined. This aim in part will examine the involvement of TRP channels in urothelial mechanotransduction. Specific Aim 2 will evaluate the mechanism by which various transmitters are released from urothelial cells. The aim will utilize imaging with fluorescent dyes to study how chemical and physical stimuli stimulate movement and release of transmitters from vesicles in urothelial cells. Specific Aim 3 will elucidate the responses evoked by mechano-stimulation in coupled and adjacent cellular targets. This aim will utilize conditions of increasing complexity (signaling within a single cell; urothelial-neuron co-cultures and intact tissue) to examine mechanisms for cell-cell interactions. These imaging methods will enable us to evaluate urothelial-cell signaling between different regions of the bladder as well as within the urothelial layers. Results from these studies will help us to understand how urothelial cells receive and integrate multiple stimuli thus providing an important "link" in the transfer of information from the urinary bladder to the nervous system. Understanding these mechanisms may provide important insight for the identification of novel targets for the future clinical management of bladder dysfunctions.

描述（申请人提供）：传统上，膀胱中的感官信号传导归因于膀胱传入的直接激活。新发现突出了膀胱尿路上皮细胞，作为感觉事件转导的关键参与者。我们已经表明，尿路上皮细胞表现出许多“神经元样”特性（包括传感器分子的表达，使它们能够对化学/热/机械刺激响应并释放化学介质的响应），使尿尿细胞可能与尿道细胞神经或间接的细胞 - 细胞相互作用“直接”“直接“直接”传达。尿路上皮细胞不断暴露于各种形式的机械刺激中，但是这些细胞响应的机制尚未得到很好的定义。人们认为机械力“启动”整个尿皮细胞中的一系列信号事件（钙波）。反过来，机械刺激引起的化学因子的随附释放可以激活尿路上皮细胞表面受体，并扩大尿路上皮细胞内部和附近的信号。尽管这些数据表明尿路上皮细胞在细胞 - 细胞信号传导中起重要作用，但这是一个相对未开发的区域，几乎没有有关感觉转导机制或细胞之间的通信方式的信息。使用涉及药理学，siRNA，发射机释放，斑块夹和新型成像技术的多学科方法，我们的目标是评估尿路上皮细胞如何接受和整合多种刺激。特定的目标1将表征尿路上皮细胞机械传感器及其机械转移途径。有证据表明，TRPV1对于尿路上皮的机械诱发的嘌呤能信号至关重要，但是尿路上皮细胞对机械力反应的机制尚未得到很好的定义。该目的部分将检查TRP通道参与尿路上皮机械转移。具体目标2将评估从尿路上皮细胞中释放出各种发射器的机制。该目的将利用荧光染料的成像来研究化学和物理刺激如何刺激尿路上皮细胞中囊泡发射器的运动和释放。具体的目标3将阐明在耦合和相邻的细胞靶标中通过机械刺激引起的响应。该目标将利用增加复杂性（单个细胞内的信号传导；尿路上神经元共培养和完整的组织）的条件来检查细胞 - 细胞相互作用的机制。这些成像方法将使我们能够评估膀胱不同区域和尿路上皮层之间的尿路细胞信号传导。这些研究的结果将有助于我们了解尿路上皮细胞如何接收和整合多种刺激，从而在信息从泌尿膀胱转移到神经系统的信息中提供了重要的“链接”。了解这些机制可能会为鉴定膀胱功能障碍的未来临床管理的新靶标提供重要的见解。

项目成果

Layer-dependent role of collagen recruitment during loading of the rat bladder wall.

• DOI: 10.1007/s10237-017-0968-5
• 发表时间: 2018-04
• 期刊: Biomechanics and modeling in mechanobiology
• 影响因子: 3.5
• 作者: Cheng F;Birder LA;Kullmann FA;Hornsby J;Watton PN;Watkins S;Thompson M;Robertson AM
• 通讯作者: Robertson AM