Measurement of gap junctional communication by fluorescence activated cell sorting.
Cell-to-cell communication via gap junctions has played a fundamental role in the orderly development of multicellular organisms. Current methods for measuring this function apply mostly to homotypic cell populations. The newly introduced Fluorescence Activated Cell Sorting (FACS) method, albeit with some limitations, is simple, reliable, and quantitative in measuring the dye transfer via gap junctions in both homotypic and heterotypic cell populations. In the homotypic setting, the result in dye transfer from the FACS method is comparable to the scrape-loading and microinjection methods. Using this FACS method, we observed a decline of cell-to-cell communication in transformed and cancer cells. We also observed a differential degree of communication between two heterotypic cell populations depending on the direction of dye transfer.
Kiang
D T
DT
Department of Medicine, University of Minnesota, Minneapolis 55455.
Kollander
R
R
Lin
H H
HH
LaVilla
S
S
Atkinson
M M
MM
D000818:Animals; D002450:Cell Communication / Q000502:physiology*; D002452:Cell Count; D002460:Cell Line; D002461:Cell Line, Transformed; D002469:Cell Separation / Q000379:methods*; D000242:Cyclic AMP / Q000502:physiology; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000502:physiology / Q000648:ultrastructure; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D017629:Gap Junctions / Q000502:physiology*; D006801:Humans; D008325:Mammary Neoplasms, Experimental / Q000473:pathology* / Q000503:physiopathology / Q000648:ultrastructure; D051379:Mice; D015203:Reproducibility of Results; D013997:Time Factors; D014407:Tumor Cells, Cultured
D005456:Fluorescent Dyes; D000242:Cyclic AMP