Granule matrix property and rapid “kiss-and-run” exocytosis contribute to the different kinetics of catecholamine release from carotid glomus and adrenal chromaffin cells at matched quantal size

2012 ◽  
Vol 90 (6) ◽  
pp. 791-801 ◽  
Author(s):  
Nan Wang ◽  
Andy K. Lee ◽  
Lei Yan ◽  
Michael R. Simpson ◽  
Amy Tse ◽  
...  

Catecholamine-containing small dense core granules (SDCGs, vesicular diameter of ∼100 nm) are prominent in carotid glomus (chemosensory) cells and some neurons, but the release kinetics from individual SDCGs has not been studied in detail. In this study, we compared the amperometric signals from glomus cells with those from adrenal chromaffin cells, which also secrete catecholamine but via large dense core granules (LDCGs, vesicular diameter of ∼200–250 nm). When exocytosis was triggered by whole-cell dialysis (which raised the concentration of intracellular Ca2+ ([Ca2+]i) to ∼0.5 µmol/L), the proportion of the type of signal that represents a flickering fusion pore was 9-fold higher for glomus cells. Yet, at the same range of quantal size (Q, the total amount of catecholamine that can be released from a granule), the kinetics of every phase of the amperometric spike signals from glomus cells was faster. Our data indicate that the last phenomenon involved at least 2 mechanisms: (i) the granule matrix of glomus cells can supply a higher concentration of free catecholamine during exocytosis; (ii) a modest elevation of [Ca2+]i triggers a form of rapid “kiss-and-run” exocytosis, which is very prevalent among glomus SDCGs and leads to incomplete release of their catecholamine content (and underestimation of their Q value).


Neuron ◽  
2019 ◽  
Vol 102 (1) ◽  
pp. 173-183.e4 ◽  
Author(s):  
Quanfeng Zhang ◽  
Bin Liu ◽  
Qihui Wu ◽  
Bing Liu ◽  
Yinglin Li ◽  
...  


1999 ◽  
Vol 341 (1) ◽  
pp. 179-183 ◽  
Author(s):  
Nathalie CHARVIN ◽  
Geoff WILLIAMS ◽  
Robert D. BURGOYNE

Doc2 is a C2-domain-containing protein that is highly expressed in the nervous system and has a constitutively expressed isoform. It has been implicated as a potential Ca2+ sensor in regulated exocytosis, and has been suggested to be associated with synaptic vesicles. To examine whether Doc2 is associated with synaptic-like microvesicles (SLMVs) or dense-core granules in neuroendocrine cells, we examined the distribution of Doc2 in subcellular fractionation of chromaffin cells of the adrenal medulla and in PC12 cells. Doc2 did not co-distribute with SLMVs from either cell type, but did appear to co-distribute with dense-core granules from PC12 cells. In contrast, it was not associated with the dense-core granules during subcellular fractionation of the adrenal medulla, and nor did it appear to be associated with endosomes, cis-Golgi or the trans-Golgi network. In contrast, Doc2 co-distributed under all conditions with a mitochondrial marker. We conclude that Doc2 is not a general component of regulated secretory vesicles, but may instead be associated with mitochondria.



1997 ◽  
Vol 8 (2) ◽  
pp. 133-140 ◽  
Author(s):  
Kevin D. Gillis ◽  
Robert H. Chow


2001 ◽  
Vol 29 (4) ◽  
pp. 467-472 ◽  
Author(s):  
R. D. Burgoyne ◽  
R. J. Fisher ◽  
M. E. Graham ◽  
L. P. Haynes ◽  
A. Morgan

The study of regulated exocytosis uniquely allows the direct measurement of intracellular membrane fusion events in real time. We have exploited this to examine factors that regulate not only the extent but also the dynamics of single fusion/release events. The general strategy used has been to assess exocytosis in transiently transfected PC12 or adrenal chromaffin cells. We aimed to design mutant constructs based on in vitro biochemistry, in some cases informed by knowledge of protein structure. Using this approach we have demonstrated an inhibitory role for the putative Rab3 effector Noc2 that requires interaction with Rab3. Using carbon-fibre amperometry on adrenal chromaffin cells, we have demonstrated regulation of the kinetics of single granule release events consistent with changes in fusion pore dynamics and switches between full fusion and ‘kiss-and-run’ fusion. These studies have demonstrated a late role for cysteine string protein in exocytosis. In addition, they have focused attention on a key role for Munc18 in the regulation of post-fusion events that affect fusion pore dynamics.



PLoS ONE ◽  
2013 ◽  
Vol 8 (9) ◽  
pp. e75845 ◽  
Author(s):  
Virginia García-Martínez ◽  
José Villanueva ◽  
Cristina J. Torregrosa-Hetland ◽  
Robert Bittman ◽  
Ashlee Higdon ◽  
...  




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