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Analysis of regulated secretion using PC12 cells
1University of California at San Diego, La Jolla, California, USA.
Current Protocols in Cell Biology
|January 30, 2008
Summary
This study details protocols for maintaining and transfecting PC12 cells, a model for neurosecretion. Assays are provided to measure secretory granule exocytosis and cargo release in neuronal cells.
Area of Science:
- Neuroscience
- Cell Biology
Background:
- PC12 cells, derived from rat adrenal medulla, are a key model for studying neurosecretion and neuronal differentiation.
- These cells possess abundant secretory granules (large dense-core vesicles) crucial for storing and releasing various biomolecules.
Purpose of the Study:
- To provide standardized protocols for the maintenance and transfection of PC12 cells.
- To describe methods for quantifying secretory granule exocytosis and cargo release.
Main Methods:
- Cell culture and transfection techniques for PC12 cells.
- Development of secretion assays using radioactive catecholamine detection.
- Application of immunochemical and chemiluminescence methods for detecting released secretory proteins.
Main Results:
- Established protocols enable reliable PC12 cell culture and genetic manipulation.
- Quantifiable assays for measuring catecholamine release and regulated secretory protein secretion were validated.
- These methods facilitate the study of calcium-regulated exocytosis in a neuronal model.
Conclusions:
- The provided protocols and assays offer a comprehensive toolkit for investigating neurosecretory mechanisms in PC12 cells.
- This resource supports research into neuronal differentiation and the regulation of secretory granule exocytosis.
