Light-Evoked Recovery from Wortmannin-Induced Inhibition of Catecholamine Secretion and Synaptic Transmission

Wortmannin (WT) is known to inhibit catecholamine (CA) secretion in chromaffin cells. This effect was found to be sensitive to UV light in experiments designed to perform simultaneous monitoring of changes in [Ca2+]i and CA secretion in perfused rat adrenal medullas. When the change in [Ca2+]i was measured using calcium green-1 (490 nm excitation), a 35-min treatment with 10 μM WT caused a 69% inhibition of CA secretion evoked by excess (30 mM) extracellular K+ and a moderate inhibition of the [Ca2+]i response. In contrast, the same treatment of fura-2-loaded cells with WT caused only an 11% inhibition of the high-K+-evoked secretion and no significant attenuation of the [Ca2+]i response. However, during interruption of fluorometry with fura-2, the inhibitory effect of WT developed at a rate similar to that exhibited in calcium green-1-loaded cells. The WT-induced inhibition of high-K+- or bradykinin-evoked secretory responses, which was otherwise irreversible, was reversed by exposing WT-treated chromaffin cells to 380-nm light. When WT was reapplied to the cells of which the secretory ability had been restored by light irradiation, the secretory response was inhibited with a time course similar to that shown during the initial treatment with WT. The photosensitive effect of WT was also demonstrated using bullfrog sympathetic ganglia in which WT-induced inhibition of synaptic transmission was reversed by irradiation with 380-nm light. These results suggest that UV light removes the inhibitory effects of WT by disrupting the covalent bond formed between WT and a target molecule which remains to be determined, although myosin light chain kinase has been reported as the target molecule in both cases examined in this study.