The Role of Metavinculin in Mediating the Response of Vascular Smooth Muscle Cells to Extracellular Matrix Stiffness

Abstract

Specifically, this research will achieve the following three objectives: 1) to characterize metavinculin distribution and expression in response to changes in substrate stiffness, 2) to evaluate the effect of structural modification of MVt on MVt-vinculin tail (Vt) heterodimer formation and on Vt’s propensities to bind F-actin and phosphoinositol 4,5 bisphosphate (PIP2) and to bundle F-actin, and 3) to determine MVt solution structure and locate MVt residues involved in actin induced MVt-Vt heterodimer. For the first objective, VSMCs will be cultured on the polyacrylamide gel based substrates with variable stiffness. The gene expression level will be quantified by RT-qPCR. The change of localization and relative population of metavinculin in VSMCs upon different stiffness will be visualized and quantified using confocal microscopy and fractionation, respectively. The immunofluorescence micrographs will be acquired using a Zeiss confocal microscope at Mercer University (Savannah Campus) Microscopy Core Facility. The PI’s lab has been granted access to this facility, and the PI’s college will provide funds to cover the user fees for this project.

Document Details

Document Type
DoD Grant Award
Publication Date
Feb 12, 2016
Source ID
W911NF1510429

Entities

People

  • Kai Shen

Organizations

  • Army Contracting Command
  • Office of the Secretary of Defense
  • Savannah State University

Tags

Readers

  • Cellular and Molecular Pathways of Apoptosis.
  • Molecular and Cellular Biochemistry
  • Research Science/Academic Research