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General anesthetics attenuate gap junction coupling in P19 cell line
Author(s) -
Wentlandt Kirsten,
Carlen Peter L.,
Kushnir Moshe,
Naus Christian C.,
ElBeheiry Hossam
Publication year - 2005
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.20577
Subject(s) - gap junction , calcein , halothane , biophysics , flow cytometry , chemistry , anesthetic , coupling (piping) , intracellular , anesthesia , medicine , materials science , biology , biochemistry , immunology , membrane , metallurgy
Abstract Gap junction communication is widespread throughout the mammalian nervous system among neurons as well as glia. We addressed the hypothesis that general anesthetics attenuate gap junction mediated coupling in P19 cell line that can differentiate into neuronal‐like cells and astrocytes and oligodendrocytes. We characterized the extent of dye coupling over time in the P19 cell line using colocalization of chlormethylbenzamido‐1,1 dioctadecyl‐3,3,3′,3′‐tetramethylindocarbocyamine (CM‐DiI) and calcein‐AM in donor and recipient cells in cocultures. After seeding, the gap junction permeant dye calcein spreads from donor to recipient cells. CM‐DiI and calcein fluorescence identified donor and recipient cells, respectively. The extent of intercellular connections was evaluated using cell counting and flow cytometry up to 2 hr after treatment. Clinically relevant concentrations of the intravenous anesthetics propofol (15 μM) and thiopental (10 μM) attenuated gap junction permeability in P19 cell cultures. In contrast, halothane, a volatile anesthetic in a concentration (0.64 mM) relevant to its free aqueous EC 50 had no effect on gap junction coupling; however, very high halothane concentrations (2.8 mM) blocked dye transfer by ∼90%. The results indicate that halothane concentrations pertinent to clinical anesthesia were unable to attenuate gap junction communication in a cell line that can express neuronal and glial gap junction proteins; however, clinically relevant concentrations of propofol and thiopental depressed gap junction coupling. © 2005 Wiley‐Liss, Inc.