ZIGIR, a Granule-Specific Zn2+ Indicator, Reveals Human Islet α Cell Heterogeneity

Abstract

Numerous mammalian cells contain abundant Zn²⁺ in their secretory granules, yet available Zn²⁺ sensors lack the desired specificity and sensitivity for imaging granular Zn²⁺. We developed a fluorescent zinc granule indicator, ZIGIR, that possesses numerous desired properties for live cell imaging, including >100-fold fluorescence enhancement, membrane permeability, and selective enrichment to acidic granules. The combined advantages endow ZIGIR with superior sensitivity and specificity for imaging granular Zn²⁺. ZIGIR enables separation of heterogenous β cells based on their insulin content and sorting of mouse islets into pure α cells and β cells. In human islets, ZIGIR facilitates sorting of endocrine cells into highly enriched α cells and β cells, reveals unexpectedly high Zn²⁺ activity in the somatostatin granule of some δ cells, and uncovers variation in the glucagon content among human α cells. We expect broad applications of ZIGIR for studying Zn²⁺ biology and Zn²⁺-rich secretory granules and for engineering β cells with high insulin content for treating diabetes.

Keywords: FACS islet cell; TIRF imaging; ZIGIR; Zinc sensor; ZnT8; alpha cell heterogeneity; beta cell; delta cell; dense core granule; zinc imaging.