Migration of transformed renal epithelial cells is regulated by K+ channel modulation of actin cytoskeleton and cell volume.
Migration of transformed renal epithelial (MDCK-F) cells depends on the polarized activity of a Ca2+-sensitive K+ channel (IK channel; Pflügers Arch 432:R87-R93, 1996). This study was aimed at elucidating the functional link between the IK channel and the actin cytoskeleton which is required for cell locomotion. We monitored migration of MDCK-F cells with video microscopy, quantified filamentous actin with phalloidin binding, and measured the intracellular Ca2+ concentration ([Ca2+]i) with the fluorescent dye fura-2/AM. We compared the effects of IK channel activation or inhibition with those of hypotonic swelling or hypertonic shrinkage. IK channel inhibition with charybdotoxin (CTX) or cell swelling (omission of up to 50 mmol/l NaCl) as well as IK channel activation with 1-ethyl-2-benzimidazolinone (1-EBIO) or cell shrinkage (addition of up to 100 mmol/l mannitol) reduce the rate of migration dose-dependently by up to 80%, i.e., to the same extent as cytochalasin D. Inhibition of migration is accompanied either by actin depolymerization (CTX and cell swelling) or by actin polymerization (1-EBIO and cell shrinkage). Changes of migration and phalloidin binding induced by CTX and cell swelling or by 1-EBIO and cell shrinkage, respectively, are linearly correlated with each other. CTX and cell swelling elicit a rise of [Ca2+]i whereas 1-EBIO and cell shrinkage induce a slight decrease of [Ca2+]i in most MDCK-F cells. Taken together IK-channel-dependent perturbations of cell volume and anisotonicity elicit virtually identical effects on migration, actin filaments and [Ca2+]i. We therefore suggest that cell volume - possibly via [Ca2+]i - is the link between IK channel activity, actin filaments and migration. We propose a model for how temporal and local changes of cell volume can support the migration of MDCK-F cells.
Schwab
A
A
Physiologisches Institut, Universität Würzburg, Röntgenring 9, D-97070 Würzburg, Germany. albrecht.schwab@mail.uni-wuerzburg.de
Schuricht
B
B
Seeger
P
P
Reinhardt
J
J
Dartsch
P C
PC
D000199:Actins / Q000502:physiology*; D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology; D002118:Calcium / Q000032:analysis; D002120:Calcium Channel Agonists / Q000494:pharmacology; D002460:Cell Line; D002465:Cell Movement; D048429:Cell Size; D018999:Charybdotoxin / Q000494:pharmacology; D003599:Cytoskeleton / Q000502:physiology*; D004285:Dogs; D004847:Epithelial Cells / Q000648:ultrastructure; D016257:Fura-2; D007668:Kidney / Q000648:ultrastructure*; D009994:Osmolar Concentration; D010590:Phalloidine / Q000378:metabolism; D015221:Potassium Channels / Q000502:physiology*; D012965:Sodium Chloride / Q000008:administration & dosage
D000199:Actins; D001562:Benzimidazoles; D002120:Calcium Channel Agonists; D015221:Potassium Channels; D018999:Charybdotoxin; D010590:Phalloidine; D012965:Sodium Chloride; C102635:1-ethyl-2-benzimidazolinone; D002118:Calcium; D016257:Fura-2