The unitary conductances and permeability sequences of the rat connexin40 (rCx40) gap junction channels to seven monovalent cations and anions were studied in rCx40-transfected neuroblastoma 2A (N2A) cell pairs using the dual whole cell recording technique. Chloride salt cation substitutions (115 mM principal salt) resulted in the following junctional maximal single channel current-voltage relationship slope conductances (γj in pS): CsCl (153), RbCl (148), KCl (142), NaCl (115), LiCl (86), TMACl (71), TEACl (63). Reversible block of the rCx40 channel was observed with TBA. Potassium anion salt γj are: Kglutamate (160), Kacetate (160), Kaspartate (158), KNO3 (157), KF (148), KCl (142), and KBr (132). Ion selectivity was verified by measuring reversal potentials for current in rCx40 gap junction channels with asymmetric salt solutions in the two electrodes and using the Goldman-Hodgkin-Katz equation to calculate relative permeabilities. The permeabilities relative to Li+ are: Cs+ (1.38), Rb+ (1.32), K+ (1.31), Na+ (1.16), TMA+ (0.53), TEA+ (0.45), TBA+ (0.03), Cl− (0.19), glutamate− (0.04), and NO3− (0.14), assuming that the monovalent anions permeate the channel by forming ion pairs with permeant monovalent cations within the pore thereby causing proportionate decreases in the channel conductance. This hypothesis can account for why the predicted increasing conductances with increasing ion mobilities in an essentially aqueous channel were not observed for anions in the rCx40 channel. The rCx40 effective channel radius is estimated to be 6.6 Å from a theoretical fit of the relationship of relative permeability and cation radius.
Monovalent Cation Permeation through the Connexin40 Gap Junction Channel : Cs, Rb, K, Na, Li, TEA, TMA, TBA, and Effects of Anions Br, Cl, F, Acetate, Aspartate, Glutamate, and NO3
Address correspondence to Dr. Dolores A. Beblo, Department of Pharmacology, SUNY Health Science Center at Syracuse, 750 East Adams Street, Syracuse, NY 13210. Fax: 315-464-8014.
The rat Cx40 transfected N2A cells were courteously provided by Dr. Eric C. Beyer, Washington University School of Medicine, St. Louis, MO 63110. We thank Mark G. Chilton for maintaining the N2A cell cultures. We wish to thank Dr. Peter R. Brink, Dept. of Physiology and Biophysics, SUNY Health Science Center, Stony Brook, NY 11794 for helpful discussions about the biophysical determinants of ionic permeability within a pore and Dr. Joseph D. Robinson for his helpful comments on the manuscript.
The research was supported by National Institute of Health grants HL-42220 and HL-45466.
Abbreviations used in this paper: N2A, neuroblastoma 2A; rCx40, rat connexin40.
Dolores A. Beblo, Richard D. Veenstra; Monovalent Cation Permeation through the Connexin40 Gap Junction Channel : Cs, Rb, K, Na, Li, TEA, TMA, TBA, and Effects of Anions Br, Cl, F, Acetate, Aspartate, Glutamate, and NO3. J Gen Physiol 1 April 1997; 109 (4): 509–522. doi: https://doi.org/10.1085/jgp.109.4.509
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