DOI: 10.1083/jcb.149.1.33. Fusion of Constitutive Membrane Traffic with the Cell Surface Observed by Evanescent Wave Microscopy

Fusion of Constitutive Membrane Traffic with the Cell Surface Observed by Evanescent Wave Microscopy

10.1083/jcb.149.1.33

Crossref journal-article

Rockefeller University Press

The Journal of Cell Biology (291)

Abstract

Monitoring the fusion of constitutive traffic with the plasma membrane has remained largely elusive. Ideally, fusion would be monitored with high spatial and temporal resolution. Recently, total internal reflection (TIR) microscopy was used to study regulated exocytosis of fluorescently labeled chromaffin granules. In this technique, only the bottom cellular surface is illuminated by an exponentially decaying evanescent wave of light. We have used a prism type TIR setup with a penetration depth of ∼50 nm to monitor constitutive fusion of vesicular stomatitis virus glycoprotein tagged with the yellow fluorescent protein. Fusion of single transport containers (TCs) was clearly observed and gave a distinct analytical signature. TCs approached the membrane, appeared to dock, and later rapidly fuse, releasing a bright fluorescent cloud into the membrane. Observation and analysis provided insight about their dynamics, kinetics, and position before and during fusion. Combining TIR and wide-field microscopy allowed us to follow constitutive cargo from the Golgi complex to the cell surface. Our observations include the following: (1) local restrained movement of TCs near the membrane before fusion; (2) apparent anchoring near the cell surface; (3) heterogeneously sized TCs fused either completely; or (4) occasionally larger tubular-vesicular TCs partially fused at their tips.

Bibliography

Toomre, D., Steyer, J. A., Keller, P., Almers, W., & Simons, K. (2000). Fusion of Constitutive Membrane Traffic with the Cell Surface Observed by Evanescent Wave Microscopy. The Journal of Cell Biology, 149(1), 33â40.

Authors 5

Derek Toomre (first)
Jürgen A. Steyer (additional)
Patrick Keller (additional)
Wolfhard Almers (additional)
Kai Simons (additional)

References 15 Referenced 129

10.1083/jcb.89.1.141 / J. Cell Biol. / Cell–substrate contacts illuminated by total internal reflection fluorescence by Axelrod (1981)
{'key': '2024062414162477700_Axelrodetal1992', 'first-page': '289', 'article-title': 'Total internal reflection fluorescence', 'volume-title': 'Topics in Fluorescence Spectrometry', 'author': 'Axelrod', 'year': '1992'} / Topics in Fluorescence Spectrometry / Total internal reflection fluorescence by Axelrod (1992)
10.1038/41213 / Nature / Cellular secretion. Now you see it, now you don't by Betz (1997)
10.1016/S0300-9084(00)80005-X / Biochimie / Physical techniques for the study of exocytosis in isolated cells by Henry (1998)
10.1083/jcb.143.6.1485 / J. Cell Biol. / Kinetic analysis of secretory protein traffic and characterization of Golgi to plasma membrane transport intermediates in living cells by Hirschberg (1998)
10.1016/S0005-2736(97)00128-4 / Biochim. Biophys. Acta / Cellular determinants of the lateral mobility of neural cell adhesion molecules by Jacobson (1997)
10.1083/jcb.140.3.659 / J. Cell Biol. / Visualization of the dynamics of synaptic vesicle and plasma membrane proteins in living axons by Nakata (1998)
10.1098/rstb.1999.0382 / Philos. Trans. R. Soc. Lond. B. Biol. Sci. / Evanescent-wave microscopya new tool to gain insight into the control of transmitter release by Oheim (1999)
10.1083/jcb.148.1.45 / J. Cell Biol. / Correlative light-electron microscopy reveals the tubular-saccular ultrastructure of carriers operating between Golgi apparatus and plasma membrane by Polishchuk (2000)
10.1126/science.272.5259.227 / Science / Protein sorting by transport vesicles by Rothman (1996)
10.1083/jcb.129.6.1559 / J. Cell Biol. / Barriers for lateral diffusion of transferrin receptor in the plasma membrane as characterized by receptor dragging by laser tweezersfence versus tether by Sako (1995)
10.1083/jcb.149.1.23 / J. Cell Biol. / Imaging constitutive exocytosis with total internal reflection fluorescence microscopy by Schmoranzer (2000)
10.1016/S0006-3495(99)77382-0 / Biophys. J. / Tracking single secretory granules in live chromaffin cells by evanescent-field fluorescence microscopy by Steyer (1999)
10.1038/41329 / Nature / Transport, docking and exocytosis of single secretory granules in live chromaffin cells by Steyer (1997)
10.1242/jcs.112.1.21 / J. Cell Sci. / Dual-color visualization of trans-Golgi network to plasma membrane traffic along microtubules in living cells by Toomre (1999)

Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 12:45 p.m.)
Deposited	1 year, 2 months ago (June 24, 2024, 10:16 a.m.)
Indexed	1 month, 3 weeks ago (July 11, 2025, 9:29 p.m.)
Issued	25 years, 5 months ago (April 3, 2000)
Published	25 years, 5 months ago (April 3, 2000)
Published Online	25 years, 5 months ago (April 3, 2000)
Published Print	25 years, 5 months ago (April 3, 2000)

Funders 0

None

BibTeX

@article{Toomre_2000, title={Fusion of Constitutive Membrane Traffic with the Cell Surface Observed by Evanescent Wave Microscopy}, volume={149}, ISSN={1540-8140}, url={http://dx.doi.org/10.1083/jcb.149.1.33}, DOI={10.1083/jcb.149.1.33}, number={1}, journal={The Journal of Cell Biology}, publisher={Rockefeller University Press}, author={Toomre, Derek and Steyer, Jürgen A. and Keller, Patrick and Almers, Wolfhard and Simons, Kai}, year={2000}, month=apr, pages={33–40} }

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