title: Connexin 43 expression on peripheral blood eosinophils: role of gap junctions in transendothelial migration.
abstract: Eosinophils circulate in the blood and are recruited in tissues during allergic inflammation. Gap junctions mediate direct communication between adjacent cells and may represent a new way of communication between immune cells distinct from communication through cytokines and chemokines. We characterized the expression of connexin (Cx)43 by eosinophils isolated from atopic individuals using RT-PCR, Western blotting, and confocal microscopy and studied the biological functions of gap junctions on eosinophils. The formation of functional gap junctions was evaluated measuring dye transfer using flow cytometry. The role of gap junctions on eosinophil transendothelial migration was studied using the inhibitor 18-a-glycyrrhetinic acid. Peripheral blood eosinophils express Cx43 mRNA and protein. Cx43 is localized not only in the cytoplasm but also on the plasma membrane. The membrane impermeable dye BCECF transferred from eosinophils to epithelial or endothelial cells following coculture in a dose and time dependent fashion. The gap junction inhibitors 18-a-glycyrrhetinic acid and octanol did not have a significant effect on dye transfer but reduced dye exit from eosinophils. The gap junction inhibitor 18-a-glycyrrhetinic acid inhibited eosinophil transendothelial migration in a dose dependent manner. Thus, eosinophils from atopic individuals express Cx43 constitutively and Cx43 may play an important role in eosinophil transendothelial migration and function in sites of inflammation.
authors.affiliation: Pulmonary Research Group, Department of Medicine, University of Alberta, 550 HMRC, Edmonton, AB, Canada T6G 2S2.
authors.forename: Harissios
authors.initials: H
authors.lastname: Vliagoftis
authors.affiliation: Pulmonary Research Group, Department of Medicine, University of Alberta, 550 HMRC, Edmonton, AB, Canada T6G 2S2.
authors.forename: Cory
authors.initials: C
authors.lastname: Ebeling
authors.affiliation: Pulmonary Research Group, Department of Medicine, University of Alberta, 550 HMRC, Edmonton, AB, Canada T6G 2S2.
authors.forename: Ramses
authors.initials: R
authors.lastname: Ilarraza
authors.affiliation: Pulmonary Research Group, Department of Medicine, University of Alberta, 550 HMRC, Edmonton, AB, Canada T6G 2S2.
authors.forename: Salahaddin
authors.initials: S
authors.lastname: Mahmudi-Azer
authors.affiliation: Pulmonary Research Group, Department of Medicine, University of Alberta, 550 HMRC, Edmonton, AB, Canada T6G 2S2.
authors.forename: Melanie
authors.initials: M
authors.lastname: Abel
authors.affiliation: Pulmonary Research Group, Department of Medicine, University of Alberta, 550 HMRC, Edmonton, AB, Canada T6G 2S2.
authors.forename: Darryl
authors.initials: D
authors.lastname: Adamko
authors.affiliation: Pulmonary Research Group, Department of Medicine, University of Alberta, 550 HMRC, Edmonton, AB, Canada T6G 2S2.
authors.forename: A Dean
authors.initials: AD
authors.lastname: Befus
authors.affiliation: Pulmonary Research Group, Department of Medicine, University of Alberta, 550 HMRC, Edmonton, AB, Canada T6G 2S2.
authors.forename: Redwan
authors.initials: R
authors.lastname: Moqbel
mesh_terms: D002460:Cell Line; D002469:Cell Separation; D004396:Coloring Agents / Q000378:metabolism; D018031:Connexin 43 / Q000097:blood* / Q000378:metabolism*; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D004804:Eosinophils / Q000166:cytology* / Q000378:metabolism*; D005452:Fluoresceins / Q000378:metabolism; D017629:Gap Junctions / Q000378:metabolism*; D006801:Humans; D018613:Microscopy, Confocal; D055806:Microvessels / Q000166:cytology; D057705:Transendothelial and Transepithelial Migration
chemical_list: D004396:Coloring Agents; D018031:Connexin 43; D005452:Fluoresceins; C043829:2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein
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