title: A Functional Assay to Assess Connexin 43-Mediated Cell-to-Cell Communication of Second Messengers in Cultured Bone Cells.
abstract: Cell-to-cell transfer of small molecules is a fundamental way by which multicellular organisms coordinate function. Recent work has highlighted the complexity of biologic responses downstream of gap junctions. As the connexin-regulated effectors are coming into focus, there is a need to develop functional assays that allow specific testing of biologically relevant second messengers. Here, we describe a modification of the classic gap junction parachute assay to assess biologically relevant molecules passed through gap junctions.
authors.affiliation: Department of Orthopaedics, University of Maryland School of Medicine, Baltimore, MD, USA.
authors.forename: Joseph P
authors.initials: JP
authors.lastname: Stains
authors.affiliation: Department of Medicine, Division of Bone and Mineral Diseases, Musculoskeletal Research Center, Washington University, 660 S. Euclid Ave., Campus Box 8301, St Louis, MO, 63110, USA. rcivitel@dom.wustl.edu.
authors.forename: Roberto
authors.initials: R
authors.lastname: Civitelli
mesh_terms: D000818:Animals; D001842:Bone and Bones / Q000166:cytology; D002450:Cell Communication / Q000502:physiology*; D002460:Cell Line; D002478:Cells, Cultured; D018031:Connexin 43 / Q000502:physiology*; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism; D057075:Enzyme Assays / Q000379:methods*; D017629:Gap Junctions / Q000502:physiology*; D049410:Luciferases, Renilla / Q000378:metabolism; D051379:Mice; D051966:Phospholipase C gamma / Q000378:metabolism; D051496:Receptor, Fibroblast Growth Factor, Type 1 / Q000378:metabolism; D015290:Second Messenger Systems / Q000502:physiology*; D014162:Transfection / Q000379:methods
chemical_list: D018031:Connexin 43; D050659:Core Binding Factor Alpha 1 Subunit; C543076:GJA1 protein, mouse; C493716:Runx2 protein, mouse; D049410:Luciferases, Renilla; C496349:Fgfr1 protein, mouse; D051496:Receptor, Fibroblast Growth Factor, Type 1; D051966:Phospholipase C gamma; C117399:Plcg1 protein, mouse
keywords: Cell-to-cell communication; Connexin 43; Gap junctions; Luciferase reporter assay; Osteoblast; Parachute assay; Transient transfection