Roles for Serotonin in Autonomic Innervation of the Lower Urinary Tract

血清素在下尿路自主神经支配中的作用

基本信息

批准号：
8734258
负责人：
Karen Elaine Ritter
金额：
$ 2.73万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-05 至 2017-09-04
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8734258
关键词：
Adult Age Apoptotic Area Autonomic ganglion Biological Models Bladder Bladder Control Bladder mucosa Cells Cleaved cell Clinical Data Coitus Data Databases Development Disease Drug or chemical Tissue Distribution Elements Enteral Enteric Nervous System Equilibrium Exhibits Fluorescence Functional disorder Ganglia Gene Expression Gene Expression Profile Gene Family Genes Genetic Transcription Genital system Genome HTR3A gene Histopathology Human In Situ Hybridization Intestines Investigation Knock-in Mouse Knock-out Knockout Mice Knowledge Lead Lower urinary tract Maintenance Measures Molecular Profiling Mus Mutant Strains Mice Nerve Fibers Neural Crest Neuraxis Neuroglia Neurons Neuropeptides Norepinephrine Output Pathology Pattern Pelvis Peripheral Peripheral Nerves Peripheral Nervous System Phenotype Physiological Play Population Postmenopause Process Proliferating Prosencephalon Receptor Gene Reporter Research Role Serotonin Serotonin Receptors 5-HT-3 Serotonin Receptors 5-HT4 Signal Transduction Sorting - Cell Movement Staining method Stains Stem cells Stress Surveys Techniques Testing Time Tissue-Specific Gene Expression Tissues Transgenes Transgenic Organisms Urinary Incontinence Urinary system Urination Urine Woman Work base caspase-3 cell type density fetal inhibitor/antagonist insight member migration mouse model mutant nerve stem cell nerve supply nervous system development neurodevelopment neurogenesis neuron development neuronal survival novel therapeutics older women postnatal progenitor protein expression public health relevance receptor receptor expression relating to nervous system research study reuptake segregation serotonin 5 receptor success transcriptome sequencing treatment strategy urine overflow incontinence urologic

项目摘要

DESCRIPTION (provided by applicant): Decades of research have demonstrated the importance of serotonin (5-HT) signaling for the appropriate development and survival of neuronal cell populations throughout the central and enteric nervous systems. Expression of 5-HT4 receptor is necessary for neural crest (NC)-derived enteric neurons to differentiate, proliferate, and renew during postnatal development. In addition, 5-HT1 receptor expression promotes development of thalamocortical neurons in the fetal forebrain in both mice and humans. Despite thorough investigation of 5-HT signaling in these areas, there have been very few studies inspecting the role of 5-HT in development of other peripheral nervous system components. Yet, there are several lines of evidence pointing to a fundamental role for 5-HT signaling in the development and maintenance of the autonomic innervation of the lower urinary tract (LUT). Public gene expression databases indicate expression of the 5-HT3 receptor in the fetal mouse LUT. Our own preliminary work validates and extends this finding by identifying expression of the 5HT3A and 5-HT3B receptor subunits in NC-derived neuronal progenitors. The physiological relevance of 5-HT signaling in the LUT is confirmed by knock-in studies in mice. Mutant mice expressing a hypersensitive form of the 5-HT3A receptor gene, Htr3a vs/vs, display a hypertrophic bladder phenotype in conjunction with urinary incontinence. Finally, intriguing clinical data has shown profound success in treating stress and urge urinary incontinence in postmenopausal women using serotonin and norepinephrine reuptake inhibitors (SNRIs). These findings prompt an investigation of the role of 5-HT signaling in the development and maintenance of LUT innervation. The hypothesis that 5-HT signaling promotes the survival, differentiation, and balance of lineages derived from NC-derived progenitors innervating the LUT will be thoroughly tested in three specific aims. In Aim 1, the spatial, temporal and cell-type specific patterns of 5-HT3 receptor subunit expression during the development and maturation of LUT autonomic innervation will be determined via in situ hybridization and immunohistochemical localization. In Aim 2, the role of the 5-HT3A receptor in the neural development of the LUT will be defined. Migratory patterns, survival and balance of neuronal lineages in the context of Htr3a loss will be investigated using an Htr3a knockout (Htr3a-/-) mouse model crossed to a transgenic Htr3a-EGFP fluorescent reporter. In Aim 3, alterations in gene expression during fetal LUT neurogenesis that occur as a result of Htr3a loss will be determined by flow-cytometric isolation of NC-derived neuronal progenitors from the LUT. Gene expression within Htr3a positive populations will be quantified by RNA-Seq analysis and differential gene expression validated by quantitative PCR. Elucidating the processes of neurogenesis in the development of the LUT is critical for identifying disease mechanisms and developing novel therapeutic strategies for the treatment of deficits of autonomic innervation and urinary incontinence.

描述（由申请人提供）：数十年的研究证明了5-羟色胺（5-HT）信号传导对整个中枢和肠神经系统中神经元细胞群体的适当发育和存活的重要性。 5-HT4受体的表达对于神经rest（NC）衍生的肠神经元是必需的，以在产后发育过程中区分，增殖和更新。此外，5-HT1受体表达促进了小鼠和人类胎儿前脑中丘脑皮质神经元的发展。尽管在这些地区进行了5-HT信号的彻底研究，但很少有研究检查5-HT在其他周围神经系统成分发展中的作用。然而，有几条证据表明5-HT信号在下泌尿道的自主神经支配（LUT）中的发展和维持中起着基本作用。公共基因表达数据库表明5-HT3受体在胎儿小鼠LUT中的表达。我们自己的初步工作通过在NC衍生的神经元祖细胞中识别5HT3A和5-HT3B受体亚基的表达来验证并扩展了这一发现。 5-HT信号在LUT中的生理相关性通过小鼠的敲门研究证实。表达5-HT3A受体基因HTR3A与/VS的过敏形式的突变小鼠与尿失禁结合表现出肥大性膀胱表型。最后，有趣的临床数据在治疗压力和使用5-羟色胺和去甲肾上腺素再摄取抑制剂（SNRIS）的绝经后妇女（SNRIS）方面取得了深远的成功。这些发现促使研究了5-HT信号在LUT神经的发展和维持中的作用。 5-HT信号传导的假设促进了从NC衍生的祖细胞中衍生的谱系的生存，分化和平衡。在AIM 1中，空间，时间和细胞类型 LUT自主神经神经神经神经神经神经神经神经神经支配期间5-HT3受体亚基表达的特定模式将通过原位杂交和免疫组织化学定位确定。在AIM 2中，将定义5-HT3A受体在LUT神经发育中的作用。将使用HTR3A敲除（HTR3A - / - ）小鼠模型研究的迁移模式，神经元谱系的生存和平衡，在HTR3A损失的背景下进行研究。在AIM 3中，由于HTR3A损失而发生的胎儿LUT神经发生过程中的基因表达改变将通过从LUT的NC衍生神经元祖细胞进行流动性分离来确定。 HTR3A阳性种群中的基因表达将通过RNA-seq分析和通过定量PCR验证的差异基因表达来量化。阐明LUT发展中神经发生过程对于鉴定疾病机制和开发新的治疗策略至关重要，以治疗自主神经神经神经支配和尿失禁的缺陷。