ABSTRACT
Confocal microscopy with Zinquin, a fluorogenic Zn(2+)-specific indicator, was used for spatially and temporally resolved measurement of Zn2+ efflux from single pancreatic beta-cells. When cells were incubated in buffer containing Zinquin, application of insulin secretagogues evoked an increase in fluorescence around the surface of the cell, indicative of detection of Zn2+ efflux from the cell. The fluorescence increases corresponded spatially and temporally with measurements of exocytosis obtained simultaneously by amperometry. When images were taken at 266-ms intervals, the detection limit for Zn2+ was approximately 0.5 microM. With this image frequency, it was possible to observe bursts of fluorescence which were interpreted as fluctuations of Zn2+ level due to exocytosis. The average intensity of these fluorescence bursts corresponded to a Zn2+ concentration of approximately 7 microM. Since insulin is co-stored with Zn2+ in secretory vesicles, it was concluded that the Zn2+ efflux corresponded to exocytosis of insulin/Zn(2+)-containing granules from the beta-cell. Exocytosis sites identified by this technique were frequently localized to one portion of the cell, indicative of active areas of release.
Subject(s)
Fluorescent Dyes , Islets of Langerhans/metabolism , Quinolones , Tosyl Compounds , Zinc/analysis , Animals , Cells, Cultured , Exocytosis , Extracellular Space/chemistry , Intracellular Fluid/chemistry , Islets of Langerhans/chemistry , Mice , Microscopy, Confocal , Microscopy, Fluorescence , Zinc/metabolismABSTRACT
Volatiles emitted byAnastrepha suspensa andAnastrepha ludens males were trapped and compared by GC-MS. Both species emitted previously reported nine-carbon alcohols, (Z)-3-nonenol and (Z,Z)-3,6-nonadienol, and epimeric lactones, anastrephin and epianastrephin. A third isomeric lactone, the macrolide suspensolide, which was previously known only fromA. suspensa is reported here for the first time in the volatiles ofA. ludens. We also report that both species released different proportions of the same three sesquiterpene hydrocarbons:α-farnesene,ß-bisabolene, andα-trans-bergamotene. Theß-bisabolene was isolated in sufficient quantity fromA. suspensa volatiles to establish its absolute configuration as the (R)-(+)-enantiomer. Some plausible biosynthetic relationships of farnesol to the isoprenoid lactones and sesquiterpenes identified in this study are discussed. Finally, we report thatA. suspensa produces the monoterpene (Z)-ß-ocimene whileA. ludens volatiles contained limonene.
