Genetic targeting of a small fluorescent zinc indicator to cell surface for monitoring zinc secretion

Daliang Li, Lin Liu, Wen Hong Li

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Numerous mammalian cells contain Zn<sup>2+</sup> in their secretory granules. During secretion, Zn<sup>2+</sup> is coreleased with granular cargos into extracellular medium so Zn<sup>2+</sup> serves as a convenient surrogate marker for tracking the dynamics of secretion. Fluorescent Zn<sup>2+</sup> sensors that can be selectively targeted to cells of interest would be invaluable tools for imaging Zn<sup>2+</sup> release in multicellular systems including tissues and live animals. Exploiting the HaloTag labeling technology and using an optimized linker, we have engineered a fluorescent Zn<sup>2+</sup> indicator that displayed a 15-fold fluorescence enhancement upon Zn<sup>2+</sup> 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 Zn<sup>2+</sup>/insulin granules and revealed heterogeneity in secretory activity along the cell membrane that was uncoupled from cellular Ca<sup>2+</sup> 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.

Original languageEnglish (US)
Pages (from-to)1054-1063
Number of pages10
JournalACS Chemical Biology
Volume10
Issue number4
DOIs
StatePublished - Apr 17 2015

Fingerprint

Zinc
Islets of Langerhans
Imaging techniques
Monitoring
Paracrine Communication
Optics and Photonics
Exocytosis
Secretory Vesicles
Insulin-Secreting Cells
Cell membranes
Photonics
Labeling
Animals
Color
Biomarkers
Fluorescence
Cells
Cell Membrane
Insulin
Tissue

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

Cite this

Genetic targeting of a small fluorescent zinc indicator to cell surface for monitoring zinc secretion. / Li, Daliang; Liu, Lin; Li, Wen Hong.

In: ACS Chemical Biology, Vol. 10, No. 4, 17.04.2015, p. 1054-1063.

Research output: Contribution to journalArticle

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