title: Use of calcium imaging for analysis of neuronal gap junction coupling.
abstract: We recently used Western blots for connexin 36 and neuronal dye coupling with neurobiotin to measure developmental decrease in neuronal gap junction coupling in cell cultures. To ask whether Ca2+ imaging also can be used to measure changes in the amount of neuronal gap junction coupling, we defined a Ca2+ coupling coefficient as the percentage of neurons with bicuculline-induced increases in intracellular Ca2+ that are suppressed by blocking gap junctions. We demonstrate in rat and mouse hypothalamic neuronal cultures that the Ca2+ coupling coefficient decreases during culture development, this decrease is prevented by manipulations that also prevent developmental decrease in neuronal gap junction coupling, and the coefficient is low in cultures lacking connexin 36. The results indicate that Ca2+ imaging is a useful tool to quantify the amount of neuronal gap junction coupling in cultures.
authors.affiliation: Department of Cell and Molecular Biology, Tulane University, New Orleans, USA.
authors.forename: Harsha
authors.initials: H
authors.lastname: Arumugam
authors.forename: Janna V
authors.initials: JV
authors.lastname: Denisova
authors.forename: Rachael L
authors.initials: RL
authors.lastname: Neve
authors.forename: Roderick A
authors.initials: RA
authors.lastname: Corriveau
authors.forename: Andrei B
authors.initials: AB
authors.lastname: Belousov
mesh_terms: D018750:6-Cyano-7-nitroquinoxaline-2,3-dione / Q000494:pharmacology; D000704:Analysis of Variance; D000777:Anesthetics / Q000494:pharmacology; D000818:Animals; D001640:Bicuculline / Q000494:pharmacology; D050882:CREB-Binding Protein / Q000378:metabolism / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D017630:Connexins / Q000172:deficiency; D003952:Diagnostic Imaging / Q000379:methods*; D004622:Embryo, Mammalian; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D018756:GABA Antagonists / Q000494:pharmacology; D017629:Gap Junctions / Q000378:metabolism*; D006034:Glycyrrhetinic Acid / Q000494:pharmacology; D006221:Halothane / Q000494:pharmacology; D007033:Hypothalamus, Middle / Q000166:cytology; D051379:Mice; D018345:Mice, Knockout; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D016194:Receptors, N-Methyl-D-Aspartate / Q000172:deficiency; D014633:Valine / Q000031:analogs & derivatives / Q000494:pharmacology; D000097004:Gap Junction delta-2 Protein
chemical_list: D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; D000777:Anesthetics; D001640:Bicuculline; D050882:CREB-Binding Protein; D002118:Calcium; D017630:Connexins; D018691:Excitatory Amino Acid Antagonists; D018756:GABA Antagonists; D006034:Glycyrrhetinic Acid; D006221:Halothane; D016194:Receptors, N-Methyl-D-Aspartate; D014633:Valine; D000097004:Gap Junction delta-2 Protein; C056639:2-amino-5-phosphopentanoic acid
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