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From the Department of Pharmacology, SUNY Health Science Center at Syracuse, Syracuse, New York 13210
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 (
j 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 j 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.
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