PC12 cells as a model for studies of regulated secretion in neuronal and endocrine cells.
Pheochromocytoma-derived cell lines such as PC12 cells maintain a differentiated neuroendocrine phenotype and have been widely used as a convenient model system for a wide variety of cell biological studies on neurotrophin action, monoamine biogenesis, protein trafficking, and secretory vesicle dynamics. This chapter reviews a number of methods that are useful for studies of the regulated dense core vesicle secretory pathway. This includes protocols for maintaining cells and preserving their phenotype. A variety of assays are discussed for monitoring secretion in intact or permeable cells and in transfected cells. Specific methods for immunocytochemical studies in permeable cells are discussed. Finally, protocols for high-efficiency PC12 cell transfections and the isolation of stably transfected cell lines are provided.
Martin
T F
TF
Department of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706, USA.
Grishanin
R N
RN
D000818:Animals; D012634:Bodily Secretions / Q000502:physiology*; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002478:Cells, Cultured / Q000166:cytology / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D008954:Models, Biological; D009419:Nerve Tissue Proteins / Q000032:analysis; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D009490:Neurosecretory Systems / Q000166:cytology / Q000378:metabolism*; D016716:PC12 Cells; D051381:Rats
D009419:Nerve Tissue Proteins