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[43] Permeabilizing mammalian cells to macromolecules

1989; Academic Press; Linguagem: Inglês

10.1016/s0076-6879(89)71046-6

1557-7988

Gary A. Weisman, Kevin D. Lustig, Ilan Friedberg, Leon Heppel,

Viral Infectious Diseases and Gene Expression in Insects

This chapter presents the procedures that increase the plasma membrane permeability of mammalian cells to normally impermeant compounds. Permeabilization methods described include treatment of cells with adenosine 5'-triphosphate (ATP), ionophores, Sendai virus, detergents, and Staphylococcus aureus α-toxin. Physical methods for altering the normal permeability barrier of cultured cells, including scrapeloading, treatment with hypertonic medium, and transient exposure to an electric field, are discussed. In addition, it is suggested that alterations in the plasma membrane permeability of cultured cells can be detected by assaying for the release of a prelabeled pool of normally impermeant metabolites or by measuring the uptake of normally impermeant probes into the cells. Fluorescent, radioactive, and colorimetric probes of various sizes can be used to examine the kinetics of channel formation and to approximate the size of membrane channels.