Effects of electroporation on the tubulin cytoskeleton and directed migration of corneal fibroblasts cultured within collagen matrices

Cell Motility and the Cytoskeleton
D G Harkin, E D Hay

Abstract

Electroporation provides a useful method for loading fibroblasts with fluorescent probes for the cytoskeleton, but the possible deleterious effects of this loading technique on cell motility are unknown. We have used conventional and confocal microscopy of living cells and immunohistochemistry to examine the migration and cytoskeleton of chick embryo corneal fibroblasts electroporated while cultured within collagen gels. Fibroblasts cultured in collagen (1 mg/ml) are successfully electroloaded (0.5-1.0 kVcm-1/960 microF in DMEM/F12/20 mM Hepes, pH 7.2) with dextran (4-150 kDa) and immunoglobulin, but subsequently display uncoordinated pseudopodia and hence are unable to migrate effectively in any one direction. The lack of directed movement is due to depolymerization of microtubules and/or a perinuclear collapse of vimentin filaments, seemingly caused by millimolar levels of Ca2+ ions derived from culture medium following electroporation. Fibroblasts loaded in a buffer which resembles intracellular fluid (< or = 10 microM Ca2+) maintain their cytoskeleton and continue to migrate, when returned to culture medium within 10 min. Using this novel approach, we have loaded fibroblasts migrating through extracellular matrix (ECM) with...Continue Reading

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·J WehlandK Weber
Dec 1, 1992·The Journal of Cell Biology·C M Regen, A F Horwitz
Oct 1, 1992·The Journal of Cell Biology·J A Theriot, T J Mitchison
Aug 1, 1991·Biophysical Journal·T Y Tsong
Oct 1, 1991·Current Opinion in Cell Biology·J Engel
Feb 1, 1991·The American Journal of Physiology·A K WilsonP De Lanerolle
Jul 1, 1991·The Journal of Cell Biology·A K WilsonP de Lanerolle
Jan 1, 1989·Cell Motility and the Cytoskeleton·M E Bilozur, E D Hay
Jan 1, 1989·Methods in Cell Biology·P L Mcneil
Jan 1, 1987·Cell Motility and the Cytoskeleton·A KupferS J Singer
Mar 15, 1986·The Biochemical Journal·D E Knight, M C Scrutton
Mar 1, 1985·Clinical and Experimental Dermatology·A Al-NajjarC Harrington
Oct 1, 1985·Analytical Biochemistry·M Marotta, G Martino
Sep 1, 1972·The Journal of Cell Biology·T Elsdale, J Bard
Mar 1, 1970·Experimental Cell Research·M AbercrombieS M Pegrum
Aug 1, 1972·Experimental Cell Research·M AbercrombieS M Pegrum
Feb 1, 1969·The British Journal of Ophthalmology·A Garner
Jun 1, 1995·Computer Methods and Programs in Biomedicine·R T AmpratwumJ L Bert
Jan 1, 1994·Annual Review of Cell Biology·A Hall
May 31, 1994·Annals of the New York Academy of Sciences·J C Weaver
Apr 15, 1994·Journal of Immunological Methods·J LukasM Strauss
Jun 7, 1994·Proceedings of the National Academy of Sciences of the United States of America·A D Bershadsky, A H Futerman
Jan 1, 1993·Methods in Enzymology·G A Neil, U Zimmermann
Jan 1, 1993·Journal of Mathematical Biology·R B Dickinson, R T Tranquillo
Dec 1, 1995·Developmental Biology·K B ArtingerM Bronner-Fraser

❮ Previous
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Citations

May 27, 2003·Journal of Cell Science·Karen L Schmeichel, Mina J Bissell
Mar 24, 2012·Cancer Cell International·Viktoria N PehlivanovaRumiana D Tzoneva
Jul 13, 2001·The Journal of Cell Biology·R A WorthylakeK Burridge
Sep 13, 2003·Differentiation; Research in Biological Diversity·Nicole JohnsonCarolyn Larabell
Jun 30, 2000·Current Biology : CB·E G TallM J Rebecchi
Feb 12, 2002·Nature Reviews. Molecular Cell Biology·Deepika Walpita, Elizabeth Hay
Nov 4, 2000·The Journal of Biological Chemistry·J M WatsonH S Earp
Jan 5, 2000·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·E D Hay
May 28, 2020·Toxins·Nisha ShresthaDaniel Fologea
Dec 19, 2006·Molecular Cancer Therapeutics·Chryso KanthouMaja Cemazar
Nov 26, 2020·ACS Nano·Philip M GraybillRafael V Davalos
Apr 4, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Anna ChoromańskaJolanta Saczko
Jul 28, 2018·Chemical Reviews·Martin P StewartKlavs F Jensen

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