Single cell transcriptome profiling to define cell types in brain nuclei controlling bladder function

整体-摘要 这个良性泌尿外科跨学科研究发展中心的目标是将 由世界级神经科学家和良性泌尿学研究人员组成的团队在贝斯以色列医院共同努力 女执事医疗中心将集体承担一项科学项目，制定一项教育 推广计划和建立中心网站，为泌尿外科提供宝贵的资源 社区。该科学项目将使用最先进的技术，单细胞RNA-seq(scRNA-seq)， 同时对数以千计的脑桥排尿神经元进行转录组分析 蓝斑中央核(PMC-LC)和中脑导水管周围灰质腹外侧区(PAGVL)。这些 大脑区域在控制膀胱充盈和排尿方面起着关键作用。由此产生的RNA表达 Profile将识别这两个关键区域的神经元亚群。要确定这些中的哪一个 亚群最致力于控制膀胱该项目将：1.确定 从PMC-LC投射到骶髓并接受PAGVL输入的亚群，以及PAGVL神经元 投射到PMC-LC并接受骶髓输入。2.确定哪些亚群改变了它们的RNA 对多尿症模型作出反应的表达模式。神经元亚群最有可能控制 将通过建立在其中选择性表达Cre重组酶的小鼠株系来研究膀胱， 使我们能够确定它们在正常小鼠和膀胱功能障碍小鼠中的功能作用(例如： 脑变性或前列腺源性慢性尿路梗阻)。该中心将补充和 通过让所有团队成员的专业知识参与教育外展来利用科学项目 旨在提高对良性泌尿系统疾病研究的概况和了解的努力。这个 科学工作将辅之以1)一个活跃的研讨会计划，邀请外部演讲者， 2)每年培训1-2名以欧洲为中心的暑期研究本科生，3)定期 为关键人员安排的内部中心实验室会议，向全体员工介绍他们的进展/问题 小组，4)在第一年资金完成后，中心调查人员和5)a)年终务虚会 为期一天的研讨会，题为‘神经病学的新前沿’，有两位BIDMC中心的参与者参加， 其他哈佛神经科学家和至少一位受邀的外部主旨演讲者将在 资金的第二年结束。静修和顶石研讨会将向良性的人广泛宣传。 泌尿科研究社区。所有这些研究和教育活动都将在 通过由行政核心主任建立和运行的中心网站广泛访问。 该网站最终将托管一个交互式的精选神经图谱和转录组数据库， 解释性材料，可供更广泛的社区使用和下载。此资源将 允许对许多良性泌尿系统疾病的神经基础进行有针对性的调查。

Overall - Abstract The goal of this Developmental Center for Interdisciplinary Research in Benign Urology is to bring together a team of world-class neuroscientists and benign urology researchers at the Beth Israel Deaconess Medical Center who will collectively undertake a scientific project, develop an educational outreach program and build a Center website in order to provide an invaluable resource for the urology community. The scientific project will use a state of the art technique, single cell RNA-seq (scRNA-seq), to perform simultaneous transcriptome profiling of thousands of neurons from the pontine micturition center-locus coeruleus (PMC-LC) and ventrolateral periaqueductal gray (PAGVL) brain regions. These brain regions play a critical role in controlling bladder filling and voiding. The resulting RNA expression profiles will identify the neuron subpopulations in these two crucial regions. To determine which of these subpopulations are most engaged in controlling the bladder the project will: 1. Determine which subpopulations project from PMC-LC to sacral cord and receive PAGVL input, and which PAGVL neurons project to PMC-LC and receive sacral cord input. 2. Determine which subpopulations alter their RNA expression patterns in response to a model of polyuria. Neuron subpopulations most likely to control the bladder will be studied by developing mouse lines which selectively express Cre recombinase in them, permitting us to define their functional role in normal mice and mice with disorders of bladder function (e.g. brain degeneration or prostate-induced chronic urethral obstruction). The Center will supplement and leverage the scientific project by engaging the expertise of all team members in educational outreach efforts designed to enhance the profile and understanding of benign urological disease research. The scientific efforts will be supplemented by 1) an active seminar program featuring invited outside speakers, 2) the training of 1 - 2 summer research undergraduates each year in uro-centric projects, 3) a regularly scheduled internal Center `lab' meeting for key personnel to present their progress/problems to the entire group, 4) an end-of-year retreat after the first year of funding is complete of Center investigators and 5) a day long symposium titled `New Frontiers in NeuroUrology' featuring both BIDMC Center participants, other Harvard neuroscientists and at least one invited external keynote speaker that will take place at the end of year 2 of funding. The retreat and the capstone symposium will be advertised broadly to the benign urology research community. All of these research and educational initiatives will be advertised on and made broadly accessible through a Center website to be built and run by the Administrative Core Director. This website will ultimately host an interactive curated neural atlas and transcriptome database with interpretive material, available for the broader community to utilize and download. This resource will permit targeted investigations of the neural underpinnings of numerous benign urological diseases.