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Related Experiment Videos

Doc2 enhances Ca2+-dependent exocytosis from PC12 cells

S Orita1, T Sasaki, R Komuro

  • 1Shionogi Institute for Medical Science, Japan.

The Journal of Biological Chemistry
|March 29, 1996
PubMed
Summary

Doc2 (Double C2) protein enhances calcium-dependent exocytosis. Overexpression of Doc2 stimulated growth hormone release from PC12 cells, while its inhibition reduced release, confirming its role in dense core vesicle exocytosis.

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Biochemistry

Background:

  • Doc2 (Double C2) is a novel protein with two C2-like domains that binds calcium and phospholipids.
  • Doc2 is highly expressed in the brain and concentrated in synaptic vesicle fractions.

Purpose of the Study:

  • To investigate the role of Doc2 in calcium-dependent exocytosis in cultured PC12 cells.
  • To determine if Doc2 influences the release of dense core vesicles.

Main Methods:

  • Utilized a growth hormone (GH) co-expression assay in PC12 cells to measure exocytosis.
  • Employed Northern and Western blot analyses to detect Doc2 expression.
  • Investigated the effects of Doc2 overexpression, antisense inhibition, and Doc2 mutants on GH release.

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Main Results:

  • Doc2 is present in PC12 cells.
  • Overexpression of Doc2 significantly stimulated Ca2+-dependent, high K+-induced GH release.
  • Inhibition of Doc2 expression or expression of Doc2 mutants impaired GH release.

Conclusions:

  • Doc2 plays a crucial role in enhancing Ca2+-dependent exocytosis of dense core vesicles from PC12 cells.
  • The findings suggest Doc2 is a key regulator of neurosecretory vesicle release.