Gap junctions in vascular smooth muscle provide a cell-to-cell conduction pathway that may contribute to regulation and coordination of changes in vascular tone. A7r5 cells, a cell line derived from embryonic rat aorta, express connexin 43 (Cx43) and connexin 40 (Cx40). Gap junction channels with three distinct unitary conductances (70, 108, and 141 pS) were observed in these cells. Events of each channel type were equally common, with an approximate frequency of 30-35%; however, the frequency at which each channel type was observed in individual cell pairs ranged between 10 and 65%. Treatment of the cells for 24-72 h with an antisense oligodeoxynucleotide (ODN) to Cx43 reduced the relative frequency of the 108- and 141-pS channel events, whereas comparable treatment with antisense Cx40 ODN reduced the frequency at which 70-pS channel events were observed. The simplest explanation of these findings is that Cx43 forms the 108- and 141-pS channels, whereas Cx40 forms the 70-pS channels in A7r5 cells.