Since the fusion pore of the secretory vesicle is resealed before complete dilation during ‘kiss-and-run’ exocytosis, their cargoes are not completely released. Although the transient fusion pore is kept open for several seconds, the precise mechanisms that control fusion pore maintenance, and their physiological significance, are not well understood. Using dual-colour TIRF (total internal reflection fluorescence) microscopy in neuroendocrine PC12 cells, we show that myosin II regulates the fusion pore dynamics during kiss-and-run exocytosis. The release kinetics of mRFP (monomeric red fluorescent protein)-tagged tPA (tissue plasminogen activator) and Venus-tagged BDNF (brain-derived neurotrophic factor), which show slower release kinetics than NPY (neuropeptide Y)–mRFP and insulin–mRFP, were prolonged by the overexpression of a wild-type form of the RLC (myosin II regulatory light chain). In contrast, overexpression of a dominant-negative form of RLC shortened the release kinetics. Using spH (synapto-pHluorin), a green fluorescent protein-based pH sensor inside the vesicles, we confirmed that the modulation of the release kinetics by myosin II is due to changes in the duration of fusion pore opening. In addition, we revealed that the amount of hormone released into the extracellular space upon stimulation was increased by overexpression of wild-type RLC. We propose that the duration of fusion pore opening is regulated by myosin II to control the amount of hormone released from a single vesicle.
Skip Nav Destination
Article navigation
August 2010
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
- PDF Icon PDF LinkEditorial Board
Research Article|
July 14 2010
Duration of fusion pore opening and the amount of hormone released are regulated by myosin II during kiss-and-run exocytosis
Ryo Aoki;
Ryo Aoki
1
*Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo 153-8902, Japan
Search for other works by this author on:
Tetsuya Kitaguchi;
Tetsuya Kitaguchi
1
†Life Function and Dynamics, ERATO, JST, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan
Search for other works by this author on:
Manami Oya;
Manami Oya
*Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo 153-8902, Japan
Search for other works by this author on:
Yu Yanagihara;
Yu Yanagihara
*Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo 153-8902, Japan
Search for other works by this author on:
Mai Sato;
Mai Sato
*Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo 153-8902, Japan
Search for other works by this author on:
Atsushi Miyawaki;
Atsushi Miyawaki
†Life Function and Dynamics, ERATO, JST, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan
‡Laboratory for Cell Function and Dynamics, Advanced Technology Development Group, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan
Search for other works by this author on:
Takashi Tsuboi
Takashi Tsuboi
2
*Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo 153-8902, Japan
2To whom correspondence should be addressed (email takatsuboi@bio.c.u-tokyo.ac.jp).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
December 02 2009
Revision Received:
May 05 2010
Accepted:
June 08 2010
Accepted Manuscript online:
June 08 2010
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2010 Biochemical Society
2010
Biochem J (2010) 429 (3): 497–504.
Article history
Received:
December 02 2009
Revision Received:
May 05 2010
Accepted:
June 08 2010
Accepted Manuscript online:
June 08 2010
Citation
Ryo Aoki, Tetsuya Kitaguchi, Manami Oya, Yu Yanagihara, Mai Sato, Atsushi Miyawaki, Takashi Tsuboi; Duration of fusion pore opening and the amount of hormone released are regulated by myosin II during kiss-and-run exocytosis. Biochem J 1 August 2010; 429 (3): 497–504. doi: https://doi.org/10.1042/BJ20091839
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.