Deciphering Dead-end Docking of Large Dense Core Vesicles in Bovine Chromaffin Cells

Overview

Journal J Neurosci

Specialty Neurology

Date 2013 Oct 25

PMID 24155316

Citations 11

Authors

Sandra Hugo

Ekta Dembla

Mahantappa Halimani

Ulf Matti

Jens Rettig

Ute Becherer

Affiliations

Soon will be listed here.

Abstract

Large dense core vesicle (LDCV) exocytosis in chromaffin cells follows a well characterized process consisting of docking, priming, and fusion. Total internal reflection fluorescence microscopy (TIRFM) studies suggest that some LDCVs, although being able to dock, are resistant to calcium-triggered release. This phenomenon termed dead-end docking has not been investigated until now. We characterized dead-end vesicles using a combination of membrane capacitance measurement and visualization of LDCVs with TIRFM. Stimulation of bovine chromaffin cells for 5 min with 6 μm free intracellular Ca2+ induced strong secretion and a large reduction of the LDCV density at the plasma membrane. Approximately 15% of the LDCVs were visible at the plasma membrane throughout experiments, indicating they were permanently docked dead-end vesicles. Overexpression of Munc18-2 or SNAP-25 reduced the fraction of dead-end vesicles. Conversely, expressing open-syntaxin increased the fraction of dead-end vesicles. These results indicate the existence of the unproductive target soluble N-ethylmaleimide-sensitive factor attachment protein receptor acceptor complex composed of 2:1 syntaxin-SNAP-25 in vivo. More importantly, they define a novel function for this acceptor complex in mediating dead-end docking.

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References

de Wit H, Walter A, Milosevic I, Gulyas-Kovacs A, Riedel D, Sorensen J . Synaptotagmin-1 docks secretory vesicles to syntaxin-1/SNAP-25 acceptor complexes. Cell. 2009; 138(5):935-46. DOI: 10.1016/j.cell.2009.07.027. View

Liu Y, Schirra C, Stevens D, Matti U, Speidel D, Hof D . CAPS facilitates filling of the rapidly releasable pool of large dense-core vesicles. J Neurosci. 2008; 28(21):5594-601. PMC: 6670619. DOI: 10.1523/JNEUROSCI.5672-07.2008. View

Bhaskar K, Shareef M, Sharma V, Shetty A, Ramamohan Y, Pant H . Co-purification and localization of Munc18-1 (p67) and Cdk5 with neuronal cytoskeletal proteins. Neurochem Int. 2003; 44(1):35-44. DOI: 10.1016/s0197-0186(03)00099-8. View

Dulubova I, Sugita S, Hill S, Hosaka M, Fernandez I, Sudhof T . A conformational switch in syntaxin during exocytosis: role of munc18. EMBO J. 1999; 18(16):4372-82. PMC: 1171512. DOI: 10.1093/emboj/18.16.4372. View

von Ruden L, Neher E . A Ca-dependent early step in the release of catecholamines from adrenal chromaffin cells. Science. 1993; 262(5136):1061-5. DOI: 10.1126/science.8235626. View

Johns L, Levitan E, Shelden E, Holz R, Axelrod D . Restriction of secretory granule motion near the plasma membrane of chromaffin cells. J Cell Biol. 2001; 153(1):177-90. PMC: 2185529. DOI: 10.1083/jcb.153.1.177. View

Perez Bay A, Ibanez L, Marengo F . Rapid recovery of releasable vesicles and formation of nonreleasable endosomes follow intense exocytosis in chromaffin cells. Am J Physiol Cell Physiol. 2007; 293(5):C1509-22. DOI: 10.1152/ajpcell.00632.2006. View

Bar-On D, Wolter S, van de Linde S, Heilemann M, Nudelman G, Nachliel E . Super-resolution imaging reveals the internal architecture of nano-sized syntaxin clusters. J Biol Chem. 2012; 287(32):27158-67. PMC: 3411058. DOI: 10.1074/jbc.M112.353250. View

Ashery U, Varoqueaux F, Voets T, Betz A, Thakur P, Koch H . Munc13-1 acts as a priming factor for large dense-core vesicles in bovine chromaffin cells. EMBO J. 2000; 19(14):3586-96. PMC: 313963. DOI: 10.1093/emboj/19.14.3586. View

10.

de Wit H, Cornelisse L, Toonen R, Verhage M . Docking of secretory vesicles is syntaxin dependent. PLoS One. 2007; 1:e126. PMC: 1762430. DOI: 10.1371/journal.pone.0000126. View

11.

Sorensen J, Wiederhold K, Muller E, Milosevic I, Nagy G, de Groot B . Sequential N- to C-terminal SNARE complex assembly drives priming and fusion of secretory vesicles. EMBO J. 2006; 25(5):955-66. PMC: 1409717. DOI: 10.1038/sj.emboj.7601003. View

12.

Tian J, Wu Z, Unzicker M, Lu L, Cai Q, Li C . The role of Snapin in neurosecretion: snapin knock-out mice exhibit impaired calcium-dependent exocytosis of large dense-core vesicles in chromaffin cells. J Neurosci. 2005; 25(45):10546-55. PMC: 1803083. DOI: 10.1523/JNEUROSCI.3275-05.2005. View

13.

Verhage M, Sorensen J . Vesicle docking in regulated exocytosis. Traffic. 2008; 9(9):1414-24. DOI: 10.1111/j.1600-0854.2008.00759.x. View

14.

Tellam J, McIntosh S, James D . Molecular identification of two novel Munc-18 isoforms expressed in non-neuronal tissues. J Biol Chem. 1995; 270(11):5857-63. DOI: 10.1074/jbc.270.11.5857. View

15.

Kaplan J . Nitrophenyl-EGTA, a photolabile chelator that selectively binds Ca2+ with high affinity and releases it rapidly upon photolysis. Proc Natl Acad Sci U S A. 1994; 91(1):187-91. PMC: 42911. DOI: 10.1073/pnas.91.1.187. View

16.

Pobbati A, Chan S, Lee I, Song H, Hong W . Structural and functional similarity between the Vgll1-TEAD and the YAP-TEAD complexes. Structure. 2012; 20(7):1135-40. DOI: 10.1016/j.str.2012.04.004. View

17.

Zilly F, Sorensen J, Jahn R, Lang T . Munc18-bound syntaxin readily forms SNARE complexes with synaptobrevin in native plasma membranes. PLoS Biol. 2006; 4(10):e330. PMC: 1570500. DOI: 10.1371/journal.pbio.0040330. View

18.

Speidel D, Bruederle C, Enk C, Voets T, Varoqueaux F, Reim K . CAPS1 regulates catecholamine loading of large dense-core vesicles. Neuron. 2005; 46(1):75-88. DOI: 10.1016/j.neuron.2005.02.019. View

19.

Weninger K, Bowen M, Choi U, Chu S, Brunger A . Accessory proteins stabilize the acceptor complex for synaptobrevin, the 1:1 syntaxin/SNAP-25 complex. Structure. 2008; 16(2):308-20. PMC: 2856644. DOI: 10.1016/j.str.2007.12.010. View

20.

Walter A, Wiederhold K, Bruns D, Fasshauer D, Sorensen J . Synaptobrevin N-terminally bound to syntaxin-SNAP-25 defines the primed vesicle state in regulated exocytosis. J Cell Biol. 2010; 188(3):401-13. PMC: 2819690. DOI: 10.1083/jcb.200907018. View