Red, yellow, green go! – a novel tool for microscopic segregation of secretory vesicle pools according to their age

2003 ◽  
Vol 31 (4) ◽  
pp. 851-856 ◽  
Author(s):  
U.K. Wiegand ◽  
R.R. Duncan ◽  
J. Greaves ◽  
R.H. Chow ◽  
M.J. Shipston ◽  
...  
Keyword(s):  
Chromaffin Cells ◽  
Fluorescence Emission ◽  
Secretory Vesicle ◽  
Live Cells ◽  
Adrenal Chromaffin Cells ◽  
Vesicle Pools ◽  
Large Dense Core Vesicles ◽  
Confocal Fluorescence ◽  
Cargo Proteins ◽  
Adrenal Chromaffin

Large dense-core vesicles (LDCVs) were labelled in cultured bovine adrenal chromaffin cells expressing fluorescent chimaeric ‘cargo’ proteins that were targeted to these secretory vesicles. When the cells were stimulated with nicotine 48 h after transduction, the fractional loss of fluorescent LDCVs was much greater than the fractional catecholamine secretion, implying selective release of newly assembled vesicles. This was confirmed using a fluorescent ‘timer’ construct that changes its fluorescence emission from green to red over several hours, and by measurement of the location and mobility of LDCVs in live cells by confocal fluorescence microscopy. Newly assembled (green) LDCVs were located mostly in peripheral regions of the cells, were virtually immobile and could be released by nicotine, but not by Ba2+; in contrast, older (red) LDCVs were centrally located and relatively mobile, and their exocytotic release was triggered by Ba2+, but not by nicotine. We describe the image restoration procedure that is necessary in order to analyse the behaviour of LDCVs labelled with this construct.

scholarly journals Two distinct secretory vesicle–priming steps in adrenal chromaffin cells

2010 ◽  
Vol 190 (6) ◽  
pp. 1067-1077 ◽  
Author(s):  
Yuanyuan Liu ◽  
Claudia Schirra ◽  
Ludwig Edelmann ◽  
Ulf Matti ◽  
JeongSeop Rhee ◽  
...  
Keyword(s):  
Chromaffin Cells ◽  
Secretory Vesicle ◽  
Adrenal Chromaffin Cells ◽  
Attachment Protein ◽  
Calcium Dependent ◽  
Open Conformation ◽  
Large Dense Core Vesicles ◽  
Priming Process ◽  
Adrenal Chromaffin ◽  
Protein Attachment

Priming of large dense-core vesicles (LDCVs) is a Ca2+-dependent step by which LDCVs enter a release-ready pool, involving the formation of the soluble N-ethyl-maleimide sensitive fusion protein attachment protein (SNAP) receptor complex consisting of syntaxin, SNAP-25, and synaptobrevin. Using mice lacking both isoforms of the calcium-dependent activator protein for secretion (CAPS), we show that LDCV priming in adrenal chromaffin cells entails two distinct steps. CAPS is required for priming of the readily releasable LDCV pool and sustained secretion in the continued presence of high Ca2+ concentrations. Either CAPS1 or CAPS2 can rescue secretion in cells lacking both CAPS isoforms. Furthermore, the deficit in the readily releasable LDCV pool resulting from CAPS deletion is reversed by a constitutively open form of syntaxin but not by Munc13-1, a priming protein that facilitates the conversion of syntaxin to the open conformation. Our data indicate that CAPS functions downstream of Munc13s but also interacts functionally with Munc13s in the LDCV-priming process.

scholarly journals Vesicle Pools: Lessons from Adrenal Chromaffin Cells

2011 ◽  
Vol 3 ◽  
Author(s):  
David R. Stevens ◽  
Claudia Schirra ◽  
Ute Becherer ◽  
Jens Rettig
Keyword(s):  
Chromaffin Cells ◽  
Adrenal Chromaffin Cells ◽  
Vesicle Pools ◽  
Adrenal Chromaffin

scholarly journals Impaired maturation of large dense-core vesicles in muted-deficient adrenal chromaffin cells

2015 ◽  
Vol 128 (7) ◽  
pp. 1365-1374 ◽  
Author(s):  
Z. Hao ◽  
L. Wei ◽  
Y. Feng ◽  
X. Chen ◽  
W. Du ◽  
...  
Keyword(s):  
Chromaffin Cells ◽  
Dense Core ◽  
Adrenal Chromaffin Cells ◽  
Dense Core Vesicles ◽  
Large Dense Core Vesicles ◽  
Adrenal Chromaffin

Morphological Observations on the Granule Types in Rabbit Extra-Adrenal Chromaffin Cells.

1975 ◽  
Vol 33 ◽  
pp. 318-319
Author(s):  
Joe A. Mascorro ◽  
Robert D. Yates
Keyword(s):  
Chromaffin Cells ◽  
Sodium Phosphate ◽  
Ganglion Cells ◽  
Ultrastructural Level ◽  
Osmic Acid ◽  
Adrenal Chromaffin Cells ◽  
Potassium Iodate ◽  
Perfusion Fluid ◽  
New Zealand White Rabbits ◽  
Adrenal Chromaffin

Extra-adrenal chromaffin organs (abdominal paraganglia) constitute rich sources of catecholamines. It is believed that these bodies contain norepinephrine exclusively. However, the present workers recently observed epinephrine type granules in para- ganglion cells. This report investigates catecholamine containing granules in rabbit paraganglia at the ultrastructural level.New Zealand white rabbits (150-170 grams) were anesthetized with 50 mg/kg Nembutal (IP) and perfused with 3% glutaraldehyde buffered with 0.2M sodium phosphate, pH 7.3. The retroperitoneal tissue blocks were removed and placed in perfusion fluid for 4 hours. The abdominal paraganglia were dissected from the blocks, diced, washed in phosphate buffer and fixed in 1% osmic acid buffered with phosphate. In other animals, the glutaraldehyde perfused tissue blocks were immersed for 1 hour in 3% glutaraldehyde/2.5% potassium iodate buffered as before. The paraganglia were then diced, separated into two vials and washed in the buffer. A portion of this tissue received osmic acid fixation.

scholarly journals Stimulatory effect of serotonin on Na+-dependent Ca2+ efflux from bovine adrenal chromaffin cells in culture

1997 ◽  
Vol 73 ◽  
pp. 226
Author(s):  
Kazuo Minakuchi ◽  
Hitoshi Houchi ◽  
Masanori Yoshizumi ◽  
Yasuko Ishimura ◽  
Kyoji Morita ◽  
...  
Keyword(s):  
Chromaffin Cells ◽  
Adrenal Chromaffin Cells ◽  
Cells In Culture ◽  
Adrenal Chromaffin
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