Zn2+ and Cd2+ increase the cyclic GMP level in PC12 cells by inhibition of the cyclic nucleotide phosphodiesterase

In the present study, the influence of the heavy metal ions Cd2+ and Zn2+ on cGMP metabolism in the neurosecretory rat pheochromocytoma (PC12) cell line has been investigated. Cadmium and zinc ions showed a concentration-dependent increase of intracellular cGMP levels as determined by radioimmunoassay: a 20-fold increase in cGMP concentration was found after 15 min of incubation with 20 muM Cd2+; and a 7-fold increase in cGMP was found after incubation with 50 muM Zn2+ (control: 6.05 +/- 2.1 pmol cGMP/mg protein). To obtain further mechanistic informations, the effects of Cd2+ and Zn2+ on intracellular 3',5'-cyclic nucleotide phosphodiesterase have been studied by a high performance liquid chromatography-based phosphodiesterase-assay. The cellular cGMP hydrolysis was found to be inhibited by these ions with an IC50 value of 6 +/- 0.7 muM for Cd2+ and 13 +/- 2.5 muM for Zn2+. Hence, dose-dependent increase in cellular cGMP content is due to an inhibition of cGMP hydrolysis and not due to an increase in cGMP synthesis. Cd2+ and Zn2+ were taken up by PC12 cells as determined by atomic absorption spectroscopy, all measurements were performed in a subtoxic concentration range. Our data illustrate that zinc and cadmium ions are efficient inhibitors of the cGMP-stimulated cyclic nucleotide PDEII in PC12 cells resulting in elevated cellular cGMP concentrations. Therefore, subtoxic doses of these metals may disturb intracellular cGMP/cAMP-signalling pathways leading to an impaired or altered gene expression. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.