Bovine adrenomedullary cells in culture have been used to study the role of myosin in vesicle transport during exocytosis. Amperometric determination of calcium-dependent catecholamine release from individual digitonin-permeabilized cells treated with 3μM wortmannin or 20mM 2,3-butanedione monoxime (BDM) and stimulated by continuous as well as repetitive calcium pulses showed alteration of slow phases of secretion when compared with control untreated cells. The specificity of these drugs for myosin inhibition was further supported by the use of peptide-18, a potent peptide affecting myosin light-chain kinase activity. These results were supported also by studying the impact of these myosin inhibitors on chromaffin granule mobility using direct visualization by dynamic confocal microscopy. Wortmannin and BDM affect drastically vesicle transport throughout the cell cytoplasm, including the region beneath the plasma membrane. Immunocytochemical studies demonstrate the presence of myosin types II and V in the cell periphery. The capability of antibodies to myosin II in abrogating the secretory response from populations of digitonin-permeabilized cells compared with the modest effect caused by anti-myosin V suggests that myosin II plays a fundamental role in the active transport of vesicles occurring in the sub-plasmalemmal area during chromaffin cell secretory activity.
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December 2002
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Research Article|
December 01 2002
The role of myosin in vesicle transport during bovine chromaffin cell secretion
Patricia ÑECO;
Patricia ÑECO
Instituto de Neurociencias, Centro Mixto CSIC-Universidad Miguel Hernández, Campus de San Juan, 03550 Alicante, Spain
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Anabel GIL;
Anabel GIL
Instituto de Neurociencias, Centro Mixto CSIC-Universidad Miguel Hernández, Campus de San Juan, 03550 Alicante, Spain
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María del MAR FRANCÉS;
María del MAR FRANCÉS
Instituto de Neurociencias, Centro Mixto CSIC-Universidad Miguel Hernández, Campus de San Juan, 03550 Alicante, Spain
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Salvador VINIEGRA;
Salvador VINIEGRA
Instituto de Neurociencias, Centro Mixto CSIC-Universidad Miguel Hernández, Campus de San Juan, 03550 Alicante, Spain
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Luis M. GUTIÉRREZ
Luis M. GUTIÉRREZ
1
Instituto de Neurociencias, Centro Mixto CSIC-Universidad Miguel Hernández, Campus de San Juan, 03550 Alicante, Spain
1To whom correspondence should be addressed (e-mail Luisguti@umh.es).
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Publisher: Portland Press Ltd
Received:
July 10 2002
Revision Received:
August 28 2002
Accepted:
September 12 2002
Accepted Manuscript online:
September 12 2002
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2002
2002
Biochem J (2002) 368 (2): 405–413.
Article history
Received:
July 10 2002
Revision Received:
August 28 2002
Accepted:
September 12 2002
Accepted Manuscript online:
September 12 2002
Citation
Patricia ÑECO, Anabel GIL, María del MAR FRANCÉS, Salvador VINIEGRA, Luis M. GUTIÉRREZ; The role of myosin in vesicle transport during bovine chromaffin cell secretion. Biochem J 1 December 2002; 368 (2): 405–413. doi: https://doi.org/10.1042/bj20021090
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