Mapping serotonin receptors in lower urinary tract innervation

描述(申请人提供)：膀胱、骨盆或泌尿生殖道底的复发性疼痛是慢性盆腔疼痛综合征的典型特征，包括间质性膀胱炎和慢性前列腺炎。尽管努力确定疾病标志物，但这些疾病的致病病理尚不清楚。支配下尿路的痛感受器神经元的细胞体位于背根节和许多其他感觉神经元之间，在炎症和感染后可能会产生不适当的活动。这些过程如何导致伤害性感受器的敏感性改变，并为慢性疼痛奠定基础，目前尚不完全清楚。在整个神经系统中，5-羟色胺(5-HT)对神经元的神经发生、分化和维持发挥着广泛的作用，并且可以作为一种炎性神经调节剂。通过5-羟色胺受体(5-HTRs)传递的信号已被证明可以调节伤害性感觉，并有助于感觉神经元的敏化。下尿路中通过5-HTRs传递信号的作用尚未被详细研究，因为在该系统中甚至还没有对这些受体的细胞类型特异性分布进行评估。这项研究计划旨在通过确定5-HTRs在下尿路发育和成熟过程中盆腔神经的分布来产生基础知识，从而增加对膀胱和尿路伤害性感受过程的了解。将获得5-羟色胺能腰部和骶部感觉神经元的转录图谱，以获得表达5-HTRs的盆腔感觉神经元的分子指纹。这一结果将建立一个框架，使5-羟色胺信号在泌尿生殖道中的作用可能与其他伤害性感受器有关。为了进一步阐明5-羟色胺信号和伤害性感受之间的联系，我们将产生在伤害性感觉神经元中表达多顺反子荧光报告蛋白的转基因报告鼠。多光谱转基因表达将能够同时定位细胞核、质膜和伤害性感受器内细胞内钙的波动。这些记者将能够直接成像细胞对药物干预和环境攻击的反应。所获得的信息将促进未来对下尿路慢性内脏疼痛的研究，从而满足纳入伤害性生殖器发育分子解剖学项目(NGUDMAP)的标准。

DESCRIPTION (provided by applicant): Recurrent pain of the bladder, pelvis or urogenital floor is a defining feature of chronic pelvic pain syndromes including Interstitial Cystitis and Chronic Prostatitis. The causative pathology of these disorders is not known despite efforts to identify disease markers. The cell bodies of pain- sensing neurons, nociceptors, that innervate the lower urinary tract reside in the dorsal root ganglia amidst many other sensory neurons and can develop inappropriate activity following inflammation and infection. How these processes lead to altered sensitivity of nociceptors and set the stage for chronic pain is not completely understood. Throughout the nervous system serotonin (5-HT) exerts a wide range of effects on neurogenesis, differentiation, and maintenance of neurons and can act as an inflammatory neuromodulator. Signaling through serotonin receptors (5-HTRs) has been shown to modulate nociception and contributes to sensitization of the sensory neurons. The effects of signaling through 5-HTRs in the lower urinary tract have not been investigated in detail because even the cell-type specific distribution of these receptors has not been evaluated in this system. This research proposal aims to generate fundamental knowledge that will increase understanding of nociceptive processes in the bladder and urethra by identifying the distribution of 5-HTRs in pelvic innervation during development and maturation of the lower urinary tract. Transcriptional profiles of serotonergic lumbar and sacral sensory neurons will be derived to obtain a molecular fingerprint of the pelvic sensory neurons that express 5-HTRs. The results will establish a framework so that role of 5- HT signaling in the urogenital tract may be related to other nociceptors. To further elaborate links between 5-HT signaling and nociception, we will generate transgenic reporter mice that express multi-cistronic fluorescent reporter proteins in nociceptive sensory neurons. Multi- spectral transgene expression will enable simultaneous localization of cell nuclei, splasma membrane, and fluctuations in intracellular calcium within nociceptors. These reporters will enable direct imaging of cellular responses to pharmacological interventions and environmental assaults. The information gained will advance future studies of chronic visceral pain in the lower urinary tract and thus meet the criteria for inclusion in the Nociceptive GenitoUrinary Development Molecular Anatomy Project (nGUDMAP).