Genetic Targeting of a Small Fluorescent Zinc Indicator to Cell Surface for Monitoring Zinc Secretion
journal contributionposted on 17.04.2015, 00:00 by Daliang Li, Lin Liu, Wen-Hong Li
Numerous mammalian cells contain Zn2+ in their secretory granules. During secretion, Zn2+ is coreleased with granular cargos into extracellular medium so Zn2+ serves as a convenient surrogate marker for tracking the dynamics of secretion. Fluorescent Zn2+ sensors that can be selectively targeted to cells of interest would be invaluable tools for imaging Zn2+ release in multicellular systems including tissues and live animals. Exploiting the HaloTag labeling technology and using an optimized linker, we have engineered a fluorescent Zn2+ indicator that displayed a 15-fold fluorescence enhancement upon Zn2+ binding while reacting efficiently with a HaloTag enzyme in a cellular environment. Two-color imaging of ZIMIR-HaloTag and a red-emitting calcium indicator in pancreatic islet beta cells demonstrated that photoactivation of a channelrhodopsin was able to induce exocytosis of Zn2+/insulin granules and revealed heterogeneity in secretory activity along the cell membrane that was uncoupled from cellular Ca2+ activity. This integrated photonic approach for imaging and controlling the release of large dense core granules provides exquisite cellular selectivity and should facilitate future studies of stimulus-secretion coupling and paracrine signaling in secretory cells.
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Zinc Indicatorsecretory activityfluorescence enhancementsecretory granulesGenetic Targetingfuture studiessecretory cellsimagingcore granulesHaloTag enzymecell membraneZnpancreatic islet beta cellsextracellular mediumphotonic approachCell SurfaceMonitoring Zinc SecretionNumerousoptimized linkermulticellular systems