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

scholarly journals Ca2+-Dependent Activator Protein for Secretion Is Critical for the Fusion of Dense-Core Vesicles with the Membrane in Calf Adrenal Chromaffin Cells

1999 ◽  
Vol 19 (17) ◽  
pp. 7375-7383 ◽  
Author(s):  
Abdeladim Elhamdani ◽  
Thomas F. J. Martin ◽  
Judith A. Kowalchyk ◽  
Cristina R. Artalejo
Keyword(s):  
Chromaffin Cells ◽  
Dense Core ◽  
Adrenal Chromaffin Cells ◽  
Dense Core Vesicles ◽  
Activator Protein ◽  
Adrenal Chromaffin

scholarly journals Trachynilysin mediates SNARE-dependent release of catecholamines from chromaffin cells via external and stored Ca2+

2000 ◽  
Vol 113 (7) ◽  
pp. 1119-1125 ◽  
Author(s):  
F.A. Meunier ◽  
C. Mattei ◽  
P. Chameau ◽  
G. Lawrence ◽  
C. Colasante ◽  
...  
Keyword(s):  
Chromaffin Cells ◽  
Motor Nerve ◽  
Neuroendocrine Cells ◽  
Dense Core ◽  
Frog Neuromuscular Junction ◽  
Dense Core Vesicles ◽  
Protein Receptor ◽  
Large Dense Core Vesicles ◽  
Quantal Transmitter Release ◽  
Bovine Chromaffin Cells

Trachynilysin, a 159 kDa dimeric protein purified from stonefish (Synanceia trachynis) venom, dramatically increases spontaneous quantal transmitter release at the frog neuromuscular junction, depleting small clear synaptic vesicles, whilst not affecting large dense core vesicles. The basis of this insensitivity of large dense core vesicles exocytosis was examined using a fluorimetric assay to determine whether the toxin could elicit catecholamine release from bovine chromaffin cells. Unlike the case of the motor nerve endings, nanomolar concentrations of trachynilysin evoked sustained Soluble N-ethylmaleimide-sensitive fusion protein Attachment Protein REceptor-dependent exocytosis of large dense core vesicles, but only in the presence of extracellular Ca2+. However, this response to trachynilysin does not rely on Ca2+ influx through voltage-activated Ca2+ channels because the secretion was only slightly affected by blockers of L, N and P/Q types. Instead, trachynilysin elicited a localized increase in intracellular fluorescence monitored with fluo-3/AM, that precisely co-localized with the increase of fluorescence resulting from caffeine-induced release of Ca2+ from intracellular stores. Moreover, depletion of the latter stores inhibited trachynilysin-induced exocytosis. Thus, the observed requirement of external Ca2+ for stimulation of large dense core vesicles exocytosis from chromaffin cells implicates plasma membrane channels that signal efflux of Ca2+ from intracellular stores. This study also suggests that the bases of exocytosis of large dense core vesicles from motor nerve terminals and neuroendocrine cells are distinct.

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.

The GTPase Rab3a is associated with large dense core vesicles in bovine chromaffin cells and rat PC12 cells

1995 ◽  
Vol 108 (4) ◽  
pp. 1639-1649 ◽  
Author(s):  
F. Darchen ◽  
J. Senyshyn ◽  
W.H. Brondyk ◽  
D.J. Taatjes ◽  
R.W. Holz ◽  
...  
Keyword(s):  
Subcellular Localization ◽  
Pc12 Cells ◽  
Immunoelectron Microscopy ◽  
Chromaffin Cells ◽  
Secretory Granules ◽  
Dense Core ◽  
Dense Core Vesicles ◽  
Large Dense Core Vesicles ◽  
Bovine Chromaffin Cells ◽  
Specific Association

Small GTPases of the rab family control intracellular vesicle traffic in eukaryotic cells. Although the molecular mechanisms underlying the activity of the Rab proteins have not been elucidated yet, it is known that the function of these proteins is dependent on their precise subcellular localization. It has been suggested that Rab3a, which is mainly expressed in neural and endocrine cells, might regulate exocytosis. Recently, direct experimental evidence supporting this hypothesis has been obtained. Consistent with such a role for Rab3a in regulated exocytosis was the previously reported specific association of Rab3a with synaptic vesicles and with secretory granules in adrenal chromaffin cells. Since the latter result, based on subcellular fractionation, has been controversial, we have re-investigated the subcellular localization of this GTP-binding protein by using a combination of morphological techniques. Bovine chromaffin cells were labelled with an affinity-purified polyclonal anti-Rab3a antibody and analyzed by confocal microcopy. Rab3a was found to colocalize partially with dopamine beta-hydroxylase, a chromaffin granule marker. In agreement with this observation, immunoelectron microscopy revealed a specific staining of chromaffin granules. In addition to large dense core vesicles, some small vesicles were labelled. To eliminate the possibility that the staining was due to a Rab3a-related protein, we investigated by immunoelectron microscopy the localization of an epitope-tagged Rab3a expressed in rat PC12 cells. Secretory granules were specifically labelled, whereas clear microvesicles were not. These results provide further evidence supporting a specific association of the GTPase Rab3a with large dense core secretory vesicles.

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