GUDMAP: Mapping molecular regionalization of cell types along the anterior-posterior axis of the urethra

项目摘要 在哺乳动物中，尿道是从身体排泄尿液的唯一途径，并且是主要的排泄途径。 下尿路感染和性传播疾病的入口点。尿道畸形是 人类最常见的出生缺陷之一，但我们对这些缺陷的分子发育的理解， 尿道比其他系统，如中枢神经系统、肠道和四肢滞后数年，甚至数十年。前向 尿道的后（A-P）轴从膀胱延伸到尿道口，尿道口在尿道的尖端处开口。 男性的阴茎或女性的阴道口和阴蒂之间。在细胞和组织水平， 尿道管沿A-P轴沿着表现出广泛的异质性。上皮结构沿该轴沿着变化， 前列腺前尿道、前列腺尿道、膜尿道和阴茎尿道的衬里各有不同的特征。 此外，形态发生过程发生在离散的A-P位置沿沿着尿生殖窦。比如说， 附属性腺，如前列腺和尿道球腺，在特定的时间从尿生殖窦中出芽。 轴向水平。肌肉括约肌也在高度局限的区域发育。尽管这种广泛的解剖学 区域化，建立这些模式的分子解剖学尚不清楚。除此之外 发育意义，细胞的A-P特性可能影响成年功能。男性和女性在 尿道A-P轴沿着的细胞类型和分子免疫学特征影响定植 尿道和膀胱被病原体感染在这里，我们提出，规范的细胞类型的身份沿着 尿生殖窦可作为A-P分区的基本发育问题。 人们已经对胃肠道中的类似过程进行了广泛研究，得出了详细的图像 肠道区域化，括约肌发育，器官位置和身份的规范，以及 控制这些器官内的细胞类型身份。在我们的GUDMAP 2项目的基础上， 背腹（D-V）轴的LUT，这项研究的目的是确定和映射的分子区域化的细胞 沿着小鼠和人尿道的A-P轴，从膀胱到尿道口。我们将使用激光 捕获显微切割以从前列腺前、前列腺、膜和阴茎分离尿道细胞 区域的小鼠和人类尿道，并使用RNAseq来确定其转录谱。然后我们将 对小鼠和人尿道上皮的切片和整体进行原位杂交比较分析 安装，我们将使用光学投影断层扫描，以地图A-P模式的基因表达的3D。我们 目标是为泌尿学研究社区提供必要的基因表达数据基础， 研究尿道沿着的细胞类型与先天性缺陷和疾病的发展之间的关系。

Project Summary In mammals, the urethra is the sole pathway for excretion of urine from the body and it is the primary point of entry for lower urinary tract infections and sexually transmitted diseases. Urethral malformations are among the most common birth defects in humans, yet our understanding of the molecular development of the urethra lags years, perhaps decades, behind other systems, such as the CNS, gut, and limbs. The anterior to posterior (A-P) axis of the urethra extends from the bladder to the urethral meatus, which opens at the tip of the penis in males or between the vaginal opening and the clitoris in females. At the cell and tissue levels, the urethral tube shows extensive heterogeneity along its A-P axis. Epithelial structure varies along this axis, with the linings of the pre-prostatic, prostatic, membranous, and penile urethra each having a distinctive character. Moreover, morphogenetic processes occur at discrete A-P positions along the urogenital sinus. For example, the accessory sex glands, such as the prostate and bulbourethral glands bud off the urogenital sinus at specific axial levels. Muscular sphincters also develop at highly localized regions. Despite this extensive anatomical regionalization, the molecular anatomy that establishes these patterns is not understood. In addition to this developmental significance, A-P identity of cells may influence adult function. Do male-female differences in cell type identity and molecular immunologic profiles along the A-P axis of the uretha influence colonization of the urethra and bladder by pathogens? Here we propose that specification of cell type identity along the urogenital sinus can be approached as a fundamental developmental problem of A-P regionalization. Analogous processes have been studied extensively in the gastrointestinal tract, resulting in a detailed picture of gut regionalization, sphincter development, specification of the positions and identities of organs, and control of cell type identities within those organs. Building on our GUDMAP2 project, which focused on the dorsoventral (D-V) axis of the LUT, this study aims to identify and map the molecular regionalization of cells along the A-P axis of the mouse and human urethra, from the bladder to the urethral meatus. We will use laser capture microdissection to isolate urethral cells from the pre-prostatic, prostatic, membranous, and penile regions of mouse and human urethra and use RNAseq to identify their transcriptional profiles. We will then carry out a comparative in situ hybridization analysis of mouse and human urethrae in sections and whole mounts, and we will use Optical Projection Tomography to map A-P patterns of gene expression in 3D. Our goal is to generate the foundation of gene expression data necessary for the urology research community to study how cell type identity along the urethra relates to development of congenital defects and to disease.