title: A novel, highly sensitive method for assessing gap junctional coupling.
abstract: To assess gap junctional intercellular communication we have developed a tracer-based methodology which is both highly sensitive and potentially adaptable for in vivo measurements. We found that injection of serotonin revealed significantly more intercellular communication than that injection of the most permeant synthetic tracer currently in use, neurobiotin. Furthermore, mechanical tracer loading steps can be replaced by transfection with human serotonin transporter and the inclusion of serotonin in the medium. Tracer and transporter are detected using immunocytochemical techniques and the presence of cells that are tracer-positive but transporter-negative indicates junctional communication. Tracer loading in vivo using transgenesis, electroporation or viral transduction to direct expression of transporter should be more easily accomplished than with mechanical loading methods.
authors: Department of Neurobiology, Harvard Medical School, 220 Longwood Avenue, Boston, MA 02115, United States.
authors: Mingli
authors: M
authors: Hou
authors: Yaqiao
authors: Y
authors: Li
authors: David L
authors: DL
authors: Paul
mesh_terms: D002450:Cell Communication / Q000502:physiology*; D004396:Coloring Agents / Q000378:metabolism; D017629:Gap Junctions / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D012701:Serotonin / Q000378:metabolism*; D050486:Serotonin Plasma Membrane Transport Proteins / Q000235:genetics; D014162:Transfection; D019076:Transgenes
chemical_list: D004396:Coloring Agents; D050486:Serotonin Plasma Membrane Transport Proteins; D012701:Serotonin
keywords: Connexin; Dye transfer; Gap junction; Serotonin
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