Analysis of regulated secretion using PC12 cells.
The catecholamine-secreting PC12 cell line derived from the rat adrenal medulla has long been considered a model system for neurosecretion and neuronal differentiation. PC12 cells contain a large number of secretory granules (otherwise known as large dense-core vesicles) for storage of small molecules, processing enzymes, neuropeptides, and peptide hormones. Secretory granule exocytosis in PC12 cells is tightly regulated by calcium and occurs in response to a secretagogue. This unit provides protocols for maintenance and transfection of PC12 cells. Several secretion assays are described to measure the release of secretory granule cargo molecules by detection of radioactive catecholamine, or by immunochemical or chemiluminescence detection of transfected regulated secretory proteins.
Taupenot
Laurent
L
University of California at San Diego, La Jolla, California, USA.
D000469:Alkaline Phosphatase / Q000235:genetics; D000818:Animals; D020013:Calcium Signaling / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods; D053379:Chromogranin A / Q000235:genetics; D003594:Cytoplasmic Granules / Q000378:metabolism; D005089:Exocytosis / Q000502:physiology; D017930:Genes, Reporter; D019382:Human Growth Hormone / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007202:Indicators and Reagents; D007553:Isotope Labeling / Q000379:methods; D008163:Luminescent Measurements; D009638:Norepinephrine / Q000378:metabolism*; D016716:PC12 Cells / Q000378:metabolism*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014316:Tritium / Q000032:analysis
D053379:Chromogranin A; D007202:Indicators and Reagents; D011993:Recombinant Fusion Proteins; D014316:Tritium; D019382:Human Growth Hormone; D000469:Alkaline Phosphatase; D009638:Norepinephrine