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Unraveling the diverse functions of the exocyst trafficking complex in vivo with novel mouse models Noemi Polgar1, Amanda J. Lee1, Josephine A. Napoli1, Brent A. Fujimoto1, Kadee-Kalia Tamashiro1, Ben Fogelgren1 1 Department of Anatomy, Biochemistry and Physiology; John A. Burns School of Medicine, University of Hawaii. Honolulu, HI Background and Objective: The exocyst protein complex is responsible for tethering subsets of secretory vesicles to certain sites on the plasma membrane. The exocyst is required for specialized exocytic pathways, such as the insulin-induced delivery of the GLUT4 glucose transporter. It has also been implicated in a variety of cellular processes, including endocytic recycling, epithelial barrier maintenance, primary cilia formation, cell autophagy, and pathogenic infection. However, investigations of the exocyst have been hampered due to the early embryonic lethality of exocyst gene knockout mice. Methods: We have generated a novel mouse model to enable in vivo studies of exocyst function via tissue-specific inactivation of Sec10, a central exocyst subunit. Using Cre recombinase mouse strains we knocked out Sec10 and the exocyst in tissues both during embryonic development and in adult mice. We utilized tdTomato reporter strains to confirm specific Cre activity, and to fate-map Sec10-knockout cells. Results: We have successfully created several tissue-specific knockout mouse lines, aiming to evaluate exocyst function in vivo in primary ciliogenesis, epithelial barrier-formation, urinary tract development, as well as GLUT4-mediated glucose uptake. Discussion and Conclusions: We believe our unique conditional Sec10 mouse line will prove to be a useful resource in investigating exocyst-mediated trafficking in disease and development. Grant support: This work was supported by grants from the National Institutes of Health [grant numbers K01DK087852, R03DK100738, P20GM103456-06A1-8293 to B.F.]; Hawaii Community Foundation [grant number 12ADVC-51347 to B.F.]; the University of Alabama at Birmingham (UAB) Hepato/Renal Fibrocystic Diseases Core Center (HRFDCC) [grant number 5P30DK074038, Pilot award to B.F.); the March of Dimes [Basil O’Connor Starter Scholar Research Award, grant number #5-FY14-56 to B.F.]; and the University of Hawaii at Manoa Research Centers in Minority Institute, BRIDGES program [grant number 5G12MD007601, Pilot award to B.F.].
