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Title

Ca2+-triggered peptide secretion in single cells imaged with green fluorescent protein and evanescent-wave microscopy

Authors
Lang, T
Wacker, I
Steyer, J
Kaether, C
Wunderlich, I
Gerdes, H H
Almers, W
Published in Neuron. 1997, vol. 18, no. 6, p. 857-63
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 beta-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.
Keywords AnimalsCalcium/physiologyCell DegranulationChromogranins/secretionDopamine beta-Hydroxylase/metabolismGreen Fluorescent ProteinsHumansLuminescent Proteins/diagnostic useMicroscopy, Fluorescence/methodsNeurites/metabolismNeuropeptide Y/secretionPC12 CellsPeptides/secretionRatsRecombinant Fusion ProteinsVideo Recording
Identifiers
PMID: 9208853
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LANG, T et al. Ca2+-triggered peptide secretion in single cells imaged with green fluorescent protein and evanescent-wave microscopy. In: Neuron, 1997, vol. 18, n° 6, p. 857-63. https://archive-ouverte.unige.ch/unige:18931

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Deposited on : 2012-03-20

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