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Methods Mol Biol. 2016;1437:193-201. doi: 10.1007/978-1-4939-3664-9_14.

A Functional Assay to Assess Connexin 43-Mediated Cell-to-Cell Communication of Second Messengers in Cultured Bone Cells.

Author information

1
Department of Orthopaedics, University of Maryland School of Medicine, Baltimore, MD, USA.
2
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.

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.

KEYWORDS:

Cell-to-cell communication; Connexin 43; Gap junctions; Luciferase reporter assay; Osteoblast; Parachute assay; Transient transfection

PMID:
27207296
PMCID:
PMC4959905
DOI:
10.1007/978-1-4939-3664-9_14
[Indexed for MEDLINE]
Free PMC Article

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