Aparna Tiwari

Date of Graduation

Summer 2014

Degree

Master of Science in Cell and Molecular Biology

Department

Biomedical Sciences

Committee Chair

Jianjie Wang

Abstract

It was previously demonstrated that UTP (10-5M), an agonist of P2Y2R, increases vascular permeability. However, the molecular mechanism remains to be determined. VE-cadherin, a key component of the adherens junction, is responsible for regulating endothelial permeability by forming stable endothelial cell-cell adhesion. To determine VE-cadherin responseto UTP stimulation, tyrosine phosphorylation of VE-cadherin at Y658 (pY658) was measured using Western blotting in HUVEC. There was a 16% and 19% increase in pY658 level of VE-cadherin in 5 and 15 min UTP-treated samples, respectively,compared to the control sample (n=4, p<0.005 and p<0.05).To determine whether expression level of membrane-associated VE-cadherin decreases in response to 10-5M stimulation, flow cytometry was performed. The membrane-associated and total VE-cadherin expression levels were quantified. There was no change in membrane-associated VE-cadherin expression before and after UTP (10-5M) treatment (n=3, p>>0.05). The findings demonstrate that UTP-stimulation increases pY658 levels of VE-cadherin but does not affect the expression level of membrane-associated VE-cadherin within 15 min of UTP treatment.Further study using alternative approach is needed to validate flow cytometry data.

Keywords

VE-cadherin, adherens junctions, tyrosine phosphorylation at Y658 (pY658), UTP, vascular permeability

Subject Categories

Medical Molecular Biology

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© Aparna Tiwari

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