TY - JOUR
T1 - Ca1+-triggered peptide secretion in single cells imaged with green fluorescent protein and evanescent-wave microscopy
AU - Lang, Thorsten
AU - Wacker, Irene
AU - Steyer, Jürgen
AU - Kaether, Christoph
AU - Wunderlich, Ilse
AU - Soldati, Thierry
AU - Gerdes, Hans Herman
AU - Almers, Wolfhard
PY - 1997/6
Y1 - 1997/6
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0030612594&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0030612594&partnerID=8YFLogxK
U2 - 10.1016/S0896-6273(00)80325-6
DO - 10.1016/S0896-6273(00)80325-6
M3 - Article
C2 - 9208853
AN - SCOPUS:0030612594
SN - 0896-6273
VL - 18
SP - 857
EP - 863
JO - Neuron
JF - Neuron
IS - 6
ER -