The transport of reactive intermediates in a co-cultivation system: the role of intercellular communication.

The transport of reactive intermediates through gap junctions was studied in a co-cultivation system consisting of primary chick-embryo liver cells and V79 Chinese hamster cells. The formation of gap junctions was studied by measurement of the incorporation of [3H]hypoxanthine in HGPRT deficient V79 mutant cells after co-cultivation with the hepatocytes. Under control conditions the heterologous gap junctions allowed for the transfer of [3H]hypoxanthine resulting in an average value of 13 grains/cell. Addition of the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA), inhibited this transfer in a dose related way to background values. The transfer of reactive intermediates was measured as the number of sister chromatid exchanges (SCEs) induced in the V79 cells. For both compounds tested, benzo[a]pyrene and dimethylnitrosamine, transport of reactive intermediates through gap junctions was observed. When the inhibitory effects of TPA were investigated at different time points after start of the co-cultivation, inhibition was measurable after 6 h and increased to a maximal inhibition of 50%, observed after 24 h. No effect of TPA was observed on the number of SCEs. When the metabolic cooperation deficient V79 mutant cell line MC--27, was used the effects were completely comparable to those of TPA for both compounds. The lower mutagenic effects in the MC--27 cells cannot be attributed to a lower intrinsic sensitivity for mutagens of these cells.