Ca1+-triggered peptide secretion in single cells imaged with green fluorescent protein and evanescent-wave microscopy

Thorsten Lang, Irene Wacker, Jürgen Steyer, Christoph Kaether, Ilse Wunderlich, Thierry Soldati, Hans Herman Gerdes, Wolfhard Almers

Research output: Contribution to journalArticlepeer-review

194 Scopus citations

Abstract

Green fluorescent protein fused to human chromogranin B or neuropeptide Y was expressed in PC12 cells and caused bright, punctate fluorescence. The fluorescent points colocalized with the endogenous secretory granule marker dopamine β-hydroxylase. Stimulation of live PC12 cells with elevated [K+], or of permeabilized PC12 cells with Ca2+, led to Ca2+ dependent loss of fluorescence from neurites. Ca2+ stimulated secretion of both fusion proteins equally well. In living cells, single fluorescent granules were imaged by evanescent-wave fluorescence microscopy. Granules were seen to migrate; to stop, as if trapped by plasmalemmal docking sites; and then to disappear abruptly, as if through exocytosis. Evidently, GFP fused to secreted peptides is a fluorescent marker for dense-core secretory granules and may be used for time-resolved microscopy of single granules.

Original languageEnglish (US)
Pages (from-to)857-863
Number of pages7
JournalNeuron
Volume18
Issue number6
DOIs
StatePublished - Jun 1997

ASJC Scopus subject areas

  • Neuroscience(all)

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