期刊
JOURNAL OF CELLULAR PHYSIOLOGY
卷 219, 期 1, 页码 162-172出版社
WILEY
DOI: 10.1002/jcp.21660
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资金
- NCRR NIH HHS [P41 RR001395] Funding Source: Medline
- NIGMS NIH HHS [R21 GM71768] Funding Source: Medline
Many cells respond directionally to small DC electrical fields (EFs) by an unknown mechanism, but changes in intracellular Ca2+ are widely assumed to be involved. We have used zebrafish (Danio rerio) keratocytes in an effort to understand the nature of the EF-cell interaction. We find that the adult zebrafish integument drives substantial currents outward through wounds produced by scale removal, establishing that keratocytes near the wound will experience endogenous EFs. Isolated keratocytes in culture turn toward the cathode in fields as small as 7 mV mm(-1), and the response is independent of cell size. Epidermal sheets are similarly sensitive. The frequency of intracellular Ca2+ spikes and basal Ca2+ levels were increased by EFs, but the spikes were not a necessary aspect of migration or EF response. Two-photon imaging failed to detect a pattern of gradients of Ca2+ across the lamellipodia during normal or EF-induced turning but did detect a sharp, stable Ca2+ gradient at the junction of the lamellipodium and the cell body. We conclude that gradients of Ca2+ within the lamellipodium are not required for the EF response. Immunostaining revealed an anode to cathode gradient of integrin P I during EF-induced turning, and interference with integrin function attenuated the EF response. Neither electrophoretic redistribution of membrane proteins nor asymmetric perturbations of the membrane potential appear to be involved in the EF response, and we propose a new model in which hydrodynamic forces generated by electro-osmotic water flow mediate EF-cell interactions via effects on focal adhesions.
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