Electric field-induced polarization of charged cell surface proteins does not determine the direction of galvanotaxis.
Galvanotaxis, that is, migration induced by DC electric fields, is thought to play a significant role in development and wound healing, however, the mechanisms by which extrinsic electric fields orchestrate intrinsic motility responses are unknown. Using mammalian cell lines (3T3, HeLa, and CHO cells), we tested one prevailing hypothesis, namely, that electric fields polarize charged cell surface molecules, and that these polarized molecules drive directional motility. Negatively charged sialic acids, which contribute the bulk of cell surface charge, redistribute preferentially to the surface facing the direction of motility, as measured by labeling with fluorescent wheat germ agglutinin. We treated cells with neuraminidase to remove sialic acids; as expected, this decreased total cell surface charge. We also changed cell surface charge independent of sialic acid moieties, by conjugating cationic avidin to the surface of live cells. Neuraminidase inhibited the electric field-induced directional polarization of membrane ruffling and alpha4 integrin, while avidin treatment actually reversed the directional polarization of sialic acids. Neuraminidase treatment inhibited directionality but did not alter speed of motility. Surprisingly, avidin treatment did not significantly alter either directionality or speed of motility. Thus, our results demonstrate that electric field-induced polarization of charged species indeed occurs. However, polarization of the bulk of charged cell surface proteins is neither necessary nor sufficient to cause motility, thus contradicting the second part of our hypothesis. Because neuraminidase inhibited directional motility, we also conclude that sialic acids are required constituents of some cell surface molecule(s) through which electric fields mount a polarized transmembrane response.
Department of Biological Sciences, Columbia University, New York, New York, USA.
Erik I
EI
Finkelstein
Pen-hsiu Grace
PH
Chao
Clark T
CT
Hung
Jeannette Chloë
JC
Bulinski
D000818:Animals; D001360:Avidin / Q000737:chemistry; D016466:CHO Cells; D054443:Cell Migration Assays / Q000379:methods; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D016764:Cell Polarity / Q000502:physiology*; D006224:Cricetinae; D003412:Cricetulus; D004560:Electricity; D004566:Electrodes; D005455:Fluorescent Antibody Technique / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000502:physiology*; D051379:Mice; D008858:Microscopy, Phase-Contrast / Q000379:methods; D041681:NIH 3T3 Cells; D009439:Neuraminidase / Q000737:chemistry / Q000494:pharmacology; D012794:Sialic Acids / Q000737:chemistry / Q000502:physiology
D008565:Membrane Proteins; D012794:Sialic Acids; D001360:Avidin; D009439:Neuraminidase