Titration of synaptotagmin I expression differentially regulates release of norepinephrine and neuropeptide Y.
Neuroscience
; 218: 78-88, 2012 Aug 30.
Article
em En
| MEDLINE
| ID: mdl-22609930
ABSTRACT
Synaptotagmin (syt) I is a Ca(2+) sensor that has been thought to trigger all vesicle secretion with similar mechanisms. However, given the calcium and stimulation requirements of small clear, and large dense core vesicles, we hypothesized that syt I expression differentially regulates vesicle release. Therefore, in this study, we generated multiple stable cell lines of PC12 cells that each had a different and stable level of syt I expression. We determined the functional effects of titrated syt I expression on transmitter release from the two vesicle types, and showed that the transmitters, norepinephrine (NE) and neuropeptide Y (NPY), each have a threshold level of syt I expression required for their release that is different for the two transmitter types. We used carbon fiber amperometry to measure release of NE from single vesicles, and found that release ranged from 50% to 100% in the syt I-targeted cells compared to release from control cells. We used an immunoassay to measure NPY release and found that NPY release was abolished in cells that had abolished syt I expression, but cell lines that expressed 50-60% of control levels of syt I exhibited NPY release levels comparable to release of NPY from control cells. Furthermore, the vesicle fusion pore exhibited a reduced open duration when syt I was abolished, but a longer open duration time for 50% syt I expression than control cells. Therefore, vesicles have a threshold for syt I that is required to control opening of the fusion pore, expansion, and full fusion to release large dense core proteins, but not for full fusion of the small molecules like NE.
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Base de dados:
MEDLINE
Assunto principal:
Neuropeptídeo Y
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Norepinefrina
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Vesículas Secretórias
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Sinaptotagmina I
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Neurônios
Idioma:
En
Ano de publicação:
2012
Tipo de documento:
Article