Synaptotagmin VII modulates the kinetics of dense‐core vesicle exocytosis in PC12 cells
書誌事項
- 公開日
- 2007-03-19
- 権利情報
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- http://onlinelibrary.wiley.com/termsAndConditions#vor
- DOI
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- 10.1111/j.1365-2443.2007.01070.x
- 公開者
- Wiley
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説明
<jats:p>In our previous study, we showed that PC12 cell lines stably expressing synaptotagmin (Syt) VII have greater ability to release hormones Ca<jats:sup>2+</jats:sup>‐dependently than the original PC12 cells. However, the precise molecular mechanism of the enhancement of hormone secretion by Syt VII has never been elucidated. In this study, we established a PC12 cell line that stably expresses Syt VII‐green fluorescent protein (Syt VII‐GFP) or its Ca<jats:sup>2+</jats:sup>‐binding‐site‐deficient mutant (D172N/D303N substitutions; Syt VII‐DN‐GFP), and examined the effect of Syt VII‐GFP expression on the kinetics of dense‐core vesicle exocytosis by total internal reflection fluorescence (TIRF) microscopy. Both Syt VII‐GFP and Syt VII‐DN‐GFP co‐localized well with dense‐core vesicle markers, monomeric red fluorescent protein (mRFP)‐tagged neuropeptide Y (NPY‐mRFP) and cyan fluorescent protein (CFP)‐tagged tissue plasminogen activator (tPA‐CFP). Expression of Syt VII‐GFP enhanced the number of dense‐core vesicle exocytotic events, whereas expression of Syt VII‐DN‐GFP or knockdown of Syt VII‐GFP with specific small interfering RNA (siRNA) attenuated the number of exocytotic events. Monitoring individual tPA‐CFP release events revealed that “full release” events are increased in Syt VII‐GFP‐expressing cells, but not in Syt VII‐DN‐GFP‐expressing or Syt VII‐silenced cells. Our data indicate that Syt VII modulates the kinetics of Ca<jats:sup>2+</jats:sup>‐dependent dense‐core vesicle exocytosis in neuroendocrine PC12 cells, possibly by modulating fusion pore opening.</jats:p>
収録刊行物
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- Genes to Cells
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Genes to Cells 12 (4), 511-519, 2007-03-19
Wiley