• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to project page

1997 Fiscal Year Final Research Report Summary

The role of endothelial cell on neutrophil transendothelial migration

Research Project

Project/Area Number 08671507
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Thoracic surgery
Research InstitutionAkita University

Principal Investigator

MINAMIYA Yoshihiro  Akita University School of Medicine Assistant Professor, 医学部, 講師 (30239321)

Co-Investigator(Kenkyū-buntansha) KITAMURA Michihiko  Akita University School of Medicine Associate Professor, 医学部, 助教授 (10153131)
ENOMOTO Katsuhiko  Akita University School of Medicine Professor, 医学部, 教授 (20151988)
Project Period (FY) 1996 – 1997
Keywordsneutrphil / endothelial cell / transendothelial migration / myosin light chain / myosin light chain kinase
Research Abstract

Although extravasation of neutrophils is a critical step in acute inflammation, the role of the endothelial cytoskeleton in neutrophil transmigration has not been fully investigated. We used an in vitro model of neutrophil transmigration across a monolayr of human umbilical endothelial cells (HUVEC) cultured on amniotic membrane. Human neutrophils were allowed to migrate across the HUVEC monolayr in response to a gradient leukotriene B_4 and then the number of migrated neutrophils were counted microscopically. We also followed endothelial F-actin and myosin filament formation using rhodamine-phalloidin and anti-myosin antibody staining. Myosin light chain (MLC) phosphorylation in endothelial cells was determined by immunoprecipitation of [^<32>P] labeled HUVEC with anti-myosin polyclnal antibody. Normally, neutrophil migration induced F-actin formation, myosin filament formation and MLC phosphorylation in HUVEC.When HUVEC was pretreated with the myosin light chain kinase (MLCK) inhibitor, ML-9, neutrophil migration was diminished and F-actin formation, myosin filament formation and MLC phosphorylation were inhibited. Pretreatments of HUVEC with the intracellular calcium ion chelator, bis-(O-aminophenoxyl) ethane-N,N,N', N'-tetraacetic acid acetoxymethyl ester (BAPTA/AM), and the calmodulin antagonist, trifluoperazine, had similar effects. These results indicate that a calcium/calmodulin-dependent MLCK in endothelial cells regulates neutrophil transendothelial migration.

URL: 

Published: 1999-03-16  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi