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  Microtubule-dependent transport of secretory vesicles visualized in real time with a GFP-tagged secretory protein

Wacker-Schröder, I., Kaether, C., Kromer, A., Migala, A., Almers, W., & Gerdes, H. H. (1997). Microtubule-dependent transport of secretory vesicles visualized in real time with a GFP-tagged secretory protein. Journal of Cell Science, 110(13), 1453-1463. Retrieved from http://jcs.biologists.org/cgi/content/abstract/110/13/1453.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0024-BC72-E Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0024-BC73-C
Genre: Journal Article
Alternative Title : Microtubule-dependent transport of secretory vesicles visualized in real time with a GFP-tagged secretory protein

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JCellSci_110_1997_1453.pdf (Any fulltext), 517KB
 
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 Creators:
Wacker-Schröder, Irene1, Author              
Kaether, Christoph, Author
Kromer, A., Author
Migala, Andrea2, Author              
Almers, Wolfhard3, Author              
Gerdes, Hans Herman, Author
Affiliations:
1Emeritus Group Biophysics, Max Planck Institute for Medical Research, Max Planck Society, ou_1497712              
2Department of Biomedical Optics, Max Planck Institute for Medical Research, Max Planck Society, ou_1497699              
3Department of Molecular Cell Research, Max Planck Institute for Medical Research, Max Planck Society, ou_1497703              

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 Abstract: Biosynthetic transport from the trans-Golgi network (TGN) to the plasma membrane (PM) is mediated by secretory vesicles. We analyzed secretory vesicle transport in real time using a GFP-tagged secretory protein, hCgB-GFP, consisting of human chromogranin B (hCgB) and green fluorescent protein (GFP). The fusion protein was expressed transiently in Vero cells or in a stable clone after induction with butyrate. After arrest of the biosynthetic protein transport at 20 degrees C, fluorescent hCgB-GFP colocalized with TGN38, a marker of the TGN. Subsequent release of the secretion block at 37 degrees C led to the formation of green fluorescent vesicles. Confocal analysis revealed that these vesicles were devoid of TGN38 and of Texas Red-coupled transferrin and cathepsin D, markers of the endosomal/lysosomal pathway. As determined by fluorometry and metabolic labelling hCgB-GFP was secreted from the TGN to the PM with a t(1/2) of 20-30 minutes. Video-microscope analysis of green fluorescent vesicles showed brief periods of rapid directed movement with maximal velocities of 1 microm/second. Vesicle movement occurred in all directions, centrifugal, centripetal and circumferential, and 50% of the vesicles analyzed reversed their direction of movement at least once within an observation period of 45 seconds. In the presence of nocodazole the movement of fluorescent vesicles ceased. Concomitantly, secretion of hCgB-GFP was slowed but not completely blocked. We suggest that microtubules (MT) facilitate the delivery of secretory vesicles to the PM by a stochastic transport, thereby increasing the probability for a vesicle/target membrane encounter.

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Language(s): eng - English
 Dates: 1997-02-091997-04-291997-07-01
 Publication Status: Published in print
 Pages: 11
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 Rev. Type: Peer
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Title: Journal of Cell Science
Source Genre: Journal
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Publ. Info: Cambridge, U.K. : Co. of Biologists
Pages: - Volume / Issue: 110 (13) Sequence Number: - Start / End Page: 1453 - 1463 Identifier: ISSN: 0021-9533
CoNE: https://pure.mpg.de/cone/journals/resource/954925326678