Regulation of Slo Splicing in the Urogenital Systme

描述（由申请人提供）： 博士Kevin Davies有多年的分子生物学家经验。他最近进入了一个新的研究领域，泌尿学。这个K 01应用程序将使他获得泌尿生殖生理学的基本概念的经验。然后，他将处于理想的位置，将分子生物学技术应用于泌尿学领域的基本问题，作为一名独立的研究人员。 六处阿尔伯特爱因斯坦医学院的泌尿外科是一个获得泌尿生殖生理学经验的绝佳环境。它有一个活跃的多学科研究小组，其中包括泌尿外科领域的领导者，如阿诺德梅尔曼博士和乔治基督博士。该部门进行临床和基础研究，并积极将这项工作从实验室转化为临床治疗。 为了获得泌尿生殖生理学方面的经验，本研究项目将在体内和体外研究激素对泌尿生殖平滑肌组织中Slo基因剪接的影响。我们的工作假设是平滑肌肌细胞中的SIo亚型表达受激素调节，而且，这些亚型表达的变化与泌尿生殖生理/功能障碍有关。具体来说，我们将研究激素（应激轴激素和性激素）对雌性大鼠体内Slo基因剪接的影响。将在这些动物的平滑肌组织（主动脉、结肠、膀胱和阴道）中监测Slo基因剪接的变化。Slo基因的不同剪接形式将表征其细胞和生理功能。此外，将开发一种报告基因构建体，其将允许对培养的大鼠平滑肌细胞（主动脉、结肠、膀胱和阴道）中的剪接进行真实的时间分析。将分析激素在体外系统中影响剪接的能力，以及对决定Slo剪接的信号通路的调查。 该研究项目将使戴维斯博士对泌尿生殖生理学有深入的了解，同时使他能够利用他在分子生物学方面的经验。

DESCRIPTION (provided by applicant): Dr. Kevin Davies has several years of experience as a molecular biologist. He has recently entered a new field of study, Urology. This K01 application will allow him to gain experience in the basic concepts of urogenital physiology. He will then be in an ideal position to apply the techniques of molecular biology to fundamental problems in the filed of Urology as an independent researcher. The Dept. of Urology at Albert Einstein College of Medicine is an excellent environment to gain experience in urogenital physiology. It has an active multi-disciplinary research group, which includes leaders in the filed of Urology, such as Dr, Arnold Melman and Dr. George Christ. The Dept conducts both clinical and basic research, and is active in translating this work from the laboratory to clinical therapies. In order to gain experience in urogenital physiology this research project will investigate the effect of hormones on splicing of the Slo gene in urogenital smooth muscle tissue in vivo and in vitro. Our working hypothesis is that SIo isoform expression in smooth muscle myocytes is modulated by hormones, and moreover, that these changes in isoform expression are relevant to urogenital physiology/dysfunction. Specifically we will look at the effects of hormones (stress axis hormones and sex hormones on the splicing of the Slo gene in female rats in vivo. Changes in the splicing of the Slo gene will be monitored in the smooth muscle tissue of these animals (aorta, colon, bladder and vagina). The different splice forms of the Slo gene will be characterized for their cellular and physiological function. In addition, a reporter construct will be developed which will allow real time analysis of splicing in cultured rat smooth muscle cells (aorta, colon, bladder and vagina). The ability of hormones to effect splicing in this in vitro system will be analyzed, as well as investigations into the signaling pathways that determine Slo splicing. This research project will allow Dr. Davies to develop a thorough understanding of urogenital physiology, while at the same time allowing him to utilize his experience in molecular biology.