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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 (g_j) and transjunctional voltage was well described by a two-state Boltzmann equation. The half-inactivation voltage (V₀), the ratio of minimal to maximal g_j(g_min/g_max), 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 …