Sulfhydryl reagent-induced insulin release and ⁴⁵ca⁺⁺ fluxes in syrian hamster insulinoma cells

Dispersed single cell suspensions of Syrian hamster insulinoma cells were used to study the effects of a variety of sulfhydryl-binding reagents on insulin release and ⁴⁵Ca⁺⁺ flux. Incubation of cells with several organic mercurials resulted in a rapid increase in ⁴⁵Ca⁺⁺ uptake as well as increased efflux in cells which had been prelabeled with ⁴⁵Ca⁺⁺. Concomitant with increased calcium uptake was a 4 to 5-fold increase in insulin released into the medium. Incubation with alkylating reagents such as iodoacetamide and iV-ethyl maleimide or dithiol reagents such as 5, 5′-dithiobis (2-nitrobenzoic acid) failed to stimulate either⁴⁵Ca⁺⁺ flux or insulin release. Elimination of medium calcium or preincubation of cells with iV-ethyl maleimide resulted in approximately 50% inhibition of mercurial-induced insulin release from these cells. 8-(N, N₂-diethylamino)Octyl- 3, 4, 5, -trimethoxybenzoate or a-isopropyl-a [(N-methyl-iV-homoveratryl) γ-aminopropyl]3, 4, 5′-trimethoxyphenylacetonitrite hydrochloride, agents which block potassium (40 min) -stimulated calcium flux and insulin release, failed to inhibit mercurialinduced calcium flux or insulin secretion. These results indicate that sulfhydryl-binding reagents, through their interaction with critical thiol groups, promote insulin release in these insulinoma cells by inducing changes in calcium fluxes. It is possible that these thiol groups regulate calcium metabolism and, thus, insulin release under physiological conditions.