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First published online March 12, 2004
doi: 10.1242/10.1242/jcs.00986

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Roles of microtubules, cell polarity and adhesion in electric-field-mediated motility of 3T3 fibroblasts

Erik Finkelstein^1,*, Winston Chang^1,*, P.-H. Grace Chao³, Dorota Gruber¹, Audrey Minden¹, Clark T. Hung³ and J. Chloë Bulinski^1,2,

¹ Department of Biological Sciences, Columbia University, New York, NY 10027, USA
² Department of Anatomy and Cell Biology, and Department of Pathology, Colleges of Arts and Sciences, and Physicians and Surgeons
³ Department of Biomedical Engineering, School of Engineering and Applied Sciences, Columbia University, New York, NY 10027, USA

Author for correspondence (e-mail: jcb4{at}columbia.edu)

Accepted 13 November 2003

Direct-current electric fields mediate motility (galvanotaxis) of many cell types. In 3T3 fibroblasts, electric fields increased the proportion, speed and cathodal directionality of motile cells. Analogous to fibroblasts' spontaneous migration, we initially hypothesized that reorientation of microtubule components modulates galvanotaxis. However, cells with intact microtubules did not reorient them in the field and cells without microtubules still migrated, albeit slowly, thus disproving the hypothesis. We next proposed that, in monolayers wounded and placed in an electric field, reorientation of microtubule organizing centers and stable, detyrosinated microtubules towards the wound edge is necessary and/or sufficient for migration. This hypothesis was negated because field exposure mediated migration of unoriented, cathode-facing cells and curtailed migration of oriented, anode-facing cells. This led us to propose that ablating microtubule detyrosination would not affect galvanotaxis. Surprisingly, preventing microtubule detyrosination increased motility speed, suggesting that detyrosination inhibits galvanotaxis. Microtubules might enhance adhesion/de-adhesion remodeling during galvanotaxis; thus, electric fields might more effectively mediate motility of cells poorly or dynamically attached to substrata. Consistent with this hypothesis, incompletely spread cells migrated more rapidly than fully spread cells. Also, overexpression of PAK4, a Cdc42-activated kinase that decreases adhesion, enhanced galvanotaxis speed, whereas its lack decreased speed. Thus, electric fields mediate fibroblast migration via participation of microtubules and adhesive components, but their participation differs from that during spontaneous motility.

Key words: Electric field, Cell motility, Microtubule, Adhesion

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