Regulation of Slo Splicing in the Urogenital Systme

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.