Forskolin-induced cyclic AMP signaling in single adherent bovine oviductal cells: effect of dichlorodiphenyltrichloroethane (DDT) and tris(4-chlorophenyl)methanol (TCPM)

Toxicology in Vitro : an International Journal Published in Association with BIBRA
R Pöhland, U Tiemann

Abstract

The influence of tris(4-chlorophenyl)methanol (TCPM) and dichlorodiphenyltrichloroethane (o,p'DDT) on forskolin induced cAMP signalling in single adherent bovine oviductal cells was investigated. An increase in the intracellular cAMP levels was measured indirectly by an increase in the 520/580 nm fluorescence emission ratio of the protein kinase A fluorosensor (FICRhR). FICRhR was microinjected into single cells, and the 520/580 nm fluorescence emission ratio was monitored by image cytometry with an image analysis system as a measure of intracellular cAMP concentration ([cAMP](i)). Applications of dibutyryl cAMP and forskolin caused time- and dose-dependent effects on [cAMP](i) in single oviductal cells. The addition of 16 or 32 microM TCPM or DDT for 1 h to the culture medium decreased the intracellular cAMP concentration significantly, whereas 8 microM was not able to influence the [cAMP](i). In the presence of both pesticides at 16 microM the forskolin (30 microM)-induced [cAMP](i) was significantly reduced after 1 h of incubation. It is suggested that TCPM can have the same influence compared with DDT on cells responsible for reproduction.

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