Imaging the secretory pathway: the past and future impact of live cell optical techniques.
Classically, the secretory pathway has been studied using a combination of electron microscopic, biochemical and genetic approaches. In the last 20 years with the arrival of molecular biology and epitope tagging, fluorescence microscopy has become more important than previously. Moreover, with the common availability of Green Fluorescent Protein (GFP) and confocal microscopes in the last 10 years, live cell imaging has become a major experimental approach. This review highlights the impact of the recent introduction of single-cell quantitative time-lapse imaging and photobleach techniques on the study of the secretory pathway, and the potential impact of those optical techniques which may play a significant future role in the study of the Golgi apparatus and the secretory pathway. Particular attention is paid to techniques (Fluorescence Resonance Energy Transfer, Fluorescence Correlation Spectroscopy) which can monitor protein-protein interactions in living cells.
Presley
John F
JF
McGill University, Department of Anatomy and Cell Biology, 3640 University, Montreal, QC, Canada H3A 2B2. john.presley@mcgill.ca
D000818:Animals; D004721:Endoplasmic Reticulum / Q000378:metabolism* / Q000648:ultrastructure; D036681:Fluorescence Recovery After Photobleaching; D031541:Fluorescence Resonance Energy Transfer; D006056:Golgi Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D049452:Green Fluorescent Proteins / Q000032:analysis; D008853:Microscopy; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D021381:Protein Transport; D013050:Spectrometry, Fluorescence
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins