Drop-seq Project: Project

Drop-seq 项目：项目

• 批准号: 9656579
• 负责人: Mark L. Zeidel
• 金额: $ 24.69万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9656579
• 关键词: Area; Atlases; Benign; Bladder; Bladder Control; Bladder Dysfunction; Brain; Brain Injuries; Brain region; Catalogs; Cell Nucleus; Cell physiology; Cells; Communities; Complex; Defect; Dementia; Disease; Dissociation; Electrodes; Elements; Expression Profiling; Fire - disasters; Follow-Up Studies; Frequencies; Genetic Transcription; Glutamates; Hypothalamic structure; Incontinence; Knowledge; Label; Lateral; Link; Maps; Medical Care Costs; Methods; Modernization; Monitor; Mus; Neurons; Neurophysiology - biologic function; Neurotransmitters; Organ; Overactive Bladder; Pathway interactions; Pattern; Physiological; Physiology; Polyuria; Pontine structure; Population; Publishing; Reporting; Research; Signal Transduction; Specificity; Stimulus; Stretching; Stroke; Symptoms; Tissues; Transgenic Organisms; Translating; Update; Urinary Incontinence; Urination; Urine; Urologic Diseases; Work; adeno-associated viral vector; base; bladder pain; cell type; droplet sequencing; improved; in vivo; interest; locus ceruleus structure; lower urinary tract symptoms; median eminence; member; midbrain central gray substance; mind control; mouse model; neural circuit; neuroregulation; noradrenergic; relating to nervous system; response; single-cell RNA sequencing; symptom treatment; tool; transcriptome; transcriptomics; urologic; web site

Project - Abstract Millions of people suffer from urinary incontinence, urgency, frequency and bladder pain; conditions which are often grouped under the catchall of lower urinary tract symptoms (LUTS). Little is known of the underlying mechanisms that cause these symptoms, but disruptions to brain function as seen in dementia, stroke and brain injury often result in LUTS. Neural circuit defects may in fact contribute to or cause many benign urological disorders. Neural control of urine storage and voiding is highly complex and is coordinated in specific regions of the brain. Modern transgenic “tools” enable specific neuron populations (such as glutamatergic neurons in a brain region like the ventrolateral periaqueductal gray, PAGVLGLUT neurons) to be mapped, monitored, and manipulated in vivo. While these approaches represent an enormous advance in our ability to understand how particular brain circuits control organ function, we still lack the ability to identify and work with specific subpopulations of neurons which have discrete functions. For example, although PAGVLGLUT neurons share a major neurotransmitter and are located in a discrete brain area, this group of neurons is made up of many different subpopulations, each of which has a different set of inputs, a distinct set of projections, and of course a different function. What is needed is a method to identify each of these subpopulations, and to develop approaches to target them selectively, so that we can tease apart their discrete functions as well as their connections. Single cell RNA-seq (scRNA-seq) involves dissociation of brain regions to single cells, followed by sequencing of the RNA expression pattern of each cell. This approach provides an unbiased method for defining neuron subpopulations by means of their transcriptional profiles within a heterogenous tissue. Members of our research team have used this approach both to create an atlas of neuron subpopulations from the arcuate hypothalamic nucleus and median eminence, and to deduce/validate the function of these cell types. In the proposed studies, we will create an atlas of neuron subpopulations in the pontine micturition center- locus coeruleus (PMC-LC) and the PAGVL regions, which are known to regulate bladder function. Follow- up studies will identify neural subpopulations that form the bladder-controlling neurocircuit, and those that alter significantly their RNA expression profiles in response to forced profound polyuria. Our results will guide strategies to access and manipulate genetically these discrete, functionally important neuron subpopulations. The atlas and its mapping and functional annotations will be published and will be reported with regular updates on the P20 website. The proposed studies will provide the urological research community with tools of unprecedented specificity, permitting vastly improved mapping of neural circuits controlling normal voiding, and direct means of determining how disease states alter neural function so as to cause bladder dysfunction.

项目 - 摘要 数以百万计的人患有尿失禁，紧迫性，频率和膀胱疼痛；条件 通常将其分组在较低的尿路症状（LUTS）下。对 引起这些症状的基本机制，但对脑功能的干扰如痴呆所见， 中风和脑损伤通常会导致LUTS。实际上，神经回路缺陷可能导致或引起许多 良性泌尿科疾病。尿液储存和空隙的神经控制高度复杂，是 在大脑的特定区域协调。现代转基因“工具”启用特定的神经元种群 （例如诸如腹外侧灰色，pagvlglut之类的大脑区域中的谷氨酸能神经元 神经元）要在体内映射，监测和操纵。这些方法代表 我们了解特定脑电路如何控制器官功能的能力的巨大进步，我们仍然 缺乏识别和与具有离散神经元的特定亚群的能力 功能。例如，尽管Pagvlglut神经元共享一个主要的神经递质，并且位于A 离散的大脑区域，这组神经元由许多不同的亚群组成，每个亚群都有 一组不同的输入，一组不同的项目，当然还有不同的功能。需要的是 识别这些亚群中的每一个的方法，并开发方法以选择性地针对它们，因此 我们可以教导他们的离散功能及其连接。单细胞RNA-seq （SCRNA-SEQ）涉及将脑区域解离为单个细胞，然后对RNA进行测序 每个单元的表达模式。这种方法提供了定义神经元的公正方法 亚群通过其在异源组织中的转录曲线。我们的成员 研究团队已经使用这种方法来创建来自弧形的神经元亚群的地图集 下丘脑核和中位数的隆起，并推断/验证这些细胞类型的功能。在 拟议的研究，我们将在庞蒂定排尿中心中创建一个神经元亚群的地图 层层（PMC-LC）和PAGVL区域，已知可以调节膀胱功能。跟随- UP研究将确定构成膀胱控制神经循环的神经亚群，以及那些 响应强迫深层多尿液而显着改变其RNA表达谱。我们的结果将会 指南访问和操纵这些离散，功能上重要的神经元的策略 亚群。地图集及其映射和功能注释将出版，将是 报告了P20网站上的定期更新。拟议的研究将提供泌尿外科 具有前所未有的特异性工具的研究社区，允许大大改善神经元的映射 控制正常空隙的电路，并直接确定疾病状态如何改变神经 功能以引起膀胱功能障碍。