[For details of research projects, staff, position availability, rotations and teaching materials click on the link "My Laboratory"]
The epithelial Tight Junctions form the gut barrier function preventing the diffusion of allergens, toxins and pathogens from the gut lumen into systemic circulation. Disruption of tight junctions and resulting endotoxemia play crucial role not only in the pathogenesis of most gastrointestinal diseases (IBD, celiac disease, colon cancer, etc) but also in other systemic diseases such as diabetes, alcoholic liver disease, sclerosing cholangitis, etc. The focus of studies in my laboratory is to understand the molecular organization of tight junctions and its regulation by protein phosphorylation-based cell signaling. Some of the right junction proteins currently addressed in my laboratory are occludin, claudins, E-cadherin and b-catenin. Signaling elements such as c-Src, PKC isoforms, PP2A and PTP1B are being studied in relation to phosphorylation of tight junction and adherens junction proteins on tyrosine, serine and threonine residues. Our recent studies (Suzuki etal 2009; Elias etal 2009) have identified a unique regulatory motif in occludin C-terminal domain. All cellular and molecular studies are conducted in close relation to understanding the pathogenesis IBD, colon cancer, alcoholic liver disease and cholangitis.
CURRENT TECHNIQUES UTILIZED
-Culture of epithelial cells on Transwell inserts and measuring transpithelial
-Measuring macromolecular flux across epithelial monolayers.
-Analysis of cell morphology by phase contrast microscopy
-Confocal microscopy to localize specific proteins in epithelial cells.
-Live cell imaging of fluorescent proteins.
-FRET and FRAP analysis to analyze protein-protein interactions and protein dynamics in live cells.
-Site directed mutagenesis.
-Construction of expression vectors, transfection and characterization.
-Cloning transfected cells.
-Immunoprecipitation and immunoblot analyses.
-Analysis of protein phosphorylation
-Identification of phosphorylation sites by mass spectrometry and point mutations.
-Breeding and maintenance of transgenic and knockout mice.
-Ischemia reperfusion-induced tissue injury.
-Chronic and acute ethanol administration
NIH R01 DK055532 (PI: R. K. Rao) ?Intestinal Mucosal Protection by Epidermal Growth Factor? 1998-2014
NIH R01 AA12307 (PI: R. K. Rao) ?Mechanism of Endotoxin Absorption in Alcoholics,? 2000-2015
Last modified 7/20/11 1:09 PM
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