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，尿路是排出体内尿液的唯一途径，也是最主要的 下尿路感染和性传播疾病的入口点。尿路畸形是 在人类最常见的出生缺陷中，我们对分子发育的了解 尿路落后于中枢神经系统、肠道和四肢等其他系统数年，甚至数十年。前面的 尿路的后(A-P)轴从膀胱延伸至尿道口，尿道口在 阴茎男性的阴茎，女性的阴茎在阴道口和阴蒂之间在细胞和组织水平上， 尿道管沿A-P轴表现出广泛的不均一性。上皮结构沿这条轴线变化， 前列腺前部、前列腺部、膜部和阴茎尿路的衬里，每一层都有不同的特征。 此外，形态发生过程发生在沿泌尿生殖窦的离散的A-P位置。例如, 副性腺，如前列腺和球状尿道腺，在特定的时间从泌尿生殖窦萌发。 轴向水平。肌肉括约肌也在高度局部化的区域发育。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还不清楚。除了这一点， 发育意义，细胞的A-P特性可能影响成体功能。男性和女性在 细胞类型鉴定和沿尿路A-P轴的分子免疫学特征影响定植 病原体对尿路和膀胱造成的伤害吗？在这里，我们建议沿 泌尿生殖窦可以作为A-P区划的一个基本发育问题。 类似的过程在胃肠道中得到了广泛的研究，从而形成了一幅详细的图像。 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。以我们的GUDMAP2项目为基础，该项目侧重于 LUT的背腹(D-V)轴，本研究旨在识别和绘制细胞的分子区划 顺着小鼠和人尿路的A-P轴，从膀胱到尿道口。我们将使用激光 显微切割法分离前列腺前部、前列腺膜和阴茎中的尿路细胞 并使用RNAseq来鉴定它们的转录图谱。到时候我们会的 小鼠和人尿路的切片和整体原位杂交比较分析 我们将使用光学投影断层成像技术在3D中绘制基因表达的A-P模式图。我们的 目标是生成泌尿外科研究社区所需的基因表达数据的基础 研究沿尿路的细胞类型特征如何与先天性缺陷的发生和疾病有关。

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.