MCB 130L Lecture 4 University of California, BerkeleyMCB 130L阅读4 加州大学伯克利分校.pptVIP

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MCB 130L Lecture 4 University of California, BerkeleyMCB 130L阅读4 加州大学伯克利分校.ppt

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MCB 130L Lecture 4 University of California, BerkeleyMCB 130L阅读4 加州大学伯克利分校

MCB 130L Lecture 4 Immunofluorescence of the Cytoskeleton Cell Biology ModuleOverview Fluorescence/Immunofluorescence Microscopy--Cytoskeleton Transfection Vital Staining Respiration Cell Signaling Cell Culturepropagation of cells outside the organism Primary cultures Cells collected directly from tissue (Harrison, 1907) Advantage: cells have been minimally modified Disadvantage: requires sacrifice of animal mortal; must be generated for each experiment heterogeneous cell population Cell lines Characterized by “immortality” A subset of cultured cells become “transformed” spontaneously Transformation of cells by expression of certain genes Derived from tumor cells (in vivo) Cell lines from different cell types have been derived Cytoskeleton Actin Structure Cell morphology and polarity Specialized cell structures such as epithelial microvilli, hair cell stereocilia, filopodia Tracks for myosin motors Cell motility Endocytosis, transport (protein, vesicles, organelles) Cytokinesis Muscle contraction Microtubules Structural cell morphology and polarity subcellular localization of organelles Tracks for kinesin and dynein motors intracellular transport (protein, vesicles, organelles) Motility cilia and flagella (specialized structures) Mitosis Mitotic spindle Drugs used in lab Taxol* Nocodazole* Latrunculin B* Tumor promoter (TPA or PMA) Taxol Isolated from pacific yew Binds and stabilizes microtubules Promotes lateral interactions between protofilaments Low dose- blocks mitosis High dose- increases polymerization actin unaffected Nocodazole Chemically synthesized Low dose--arrests mitosis High dose--rapidly depolymerizes microtubules actin unaffected Latrunculin B Isolated from red sea sponge Binds actin monomers and inhibits polymerization Causes loss of actin fibers (collapse onto nucleus) due to continued disassembly Alters microtubule morphology as well Phorbol Myristate Acetate Tumor promoter Increases frequency w/ which certain ch