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The Journal of Neuroscience, November 15, 1999, 19(22):9848-9855
Functional Properties of Channels Formed by the Neuronal Gap Junction Protein Connexin36
Midituru Srinivas1, Renato Rozental1, 2, Takashi Kojima1, Rolf Dermietzel3, Mark Mehler1, 2, Daniele F. Condorelli4, John A. Kessler1, 2, and David C. Spray1 1 Departments of Neuroscience and Neurology, Albert Einstein College of Medicine, Bronx, New York 10461, 2 Department of Internal Medicine, Federal University of Goias, 74000 Goias, Brazil, 3 Department of Neuroanatomy/Molecular Brain Research, Ruhr University, D-44780 Bochum, Germany, and 4 Institute of Biochemistry, University of Catania, 95125 Catania, Italy
The expression and functional properties of connexin36 (Cx36) were examined in two communication-deficient cell lines (N2A-neuroblastoma and PC-12 cells) transfected with Cx36 and in hippocampal neurons that express the connexin endogenously. Transfected cells expressed the expected 2.9 kb Cx36 transcript and Cx36 immunoreactivity, whereas nontransfected cells were devoid of Cx36. The relationship between steady-state junctional conductance (gj) and transjunctional voltage was well described by a two-state Boltzmann equation. The half-inactivation voltage (V0), the ratio of minimal to maximal gj (gmin/gmax), and the equivalent gating charge were ± 75 mV, 0.55, and 1.75, respectively, indicating that Cx36 exhibits very low voltage sensitivity. Conductance of single Cx36 channels measured with patch pipettes containing 130 mM CsCl was 10-15 pS (n = 15 cell pairs); despite this low unitary conductance, Cx36 channels were permeable to the dye Lucifer yellow. Hippocampal neurons expressed Cx36 both in vivo and in culture. The electrophysiological properties of channels in cultured hippocampal neurons were similar to those of the channels expressed by the transfected cell lines, and the neuronal channels were similarly permeable to Lucifer yellow. The unique combination of weak voltage sensitivity, small unitary conductance, and permeation by anions as large as second messenger molecules endows Cx36 gap junction channels with properties well suited for mediating flexible electrical and biochemical interactions between neurons.
Key words: gap junction; connexin; Cx36; electrotonic synapse; electrical coupling; hippocampus; transfection; single channels; voltage sensitivity
Copyright © 1999 Society for Neuroscience 0270-6474/99/19229848-08$05.00/0
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