LOCALIZATION AND FUNCTION OF AN EXTRACELLULAR MATRIX PROTEIN, TINAGL1, IN RENAL EPITHELIAL CELLS

Abstract

Theextracellular matrix (ECM)playsimportant rolesin cell adhesion and communication. We arestudying the localization and function of a small ECMprotein, tubulo-interstitial nephritis antigen-like protein 1 (Tinagl1),in mammalian cell lines. Previous studies conducted in zebrafish suggest that Tinagl1is imperativefor function of motile cilia; however, it is still uncertainas to how this presumed basement membrane protein could regulate cilia, and whether thisfunction has relevance inmammalian cells havingonly primary, non-motile cilia.We will useimmunostaining ofTinagl1 in mouse renal collecting duct epithelial (IMCD3) cellsto determine if it is only on basal side or if it can also be detected on cilia on the apical side. We will establishthe knockdown of tinagl1expression using siRNA methodswith measurements of mRNA andprotein levels. Using appropriate assays, we will examine iftheknockdown of Tinagl1 causes theloss of primary cilia, changes in cell viability and programmed cell death, and/or the detachment of cell-cell or cell-matrix adhesions. Thisproject will showbasic functionsof Tinagl1 in renal epithelial cell behavior, which will eventually give us moreinformation about itsanticipated rolesin cell adhesion, cell survival, and primary cilia homeostasis.