Role of K(+) channel expression in polyamine-dependent intestinal epithelial cell migration.
Polyamines are essential for cell migration during early mucosal restitution after wounding in the gastrointestinal tract. Activity of voltage-gated K(+) channels (Kv) controls membrane potential (E(m)) that regulates cytoplasmic free Ca(2+) concentration ([Ca(2+)](cyt)) by governing the driving force for Ca(2+) influx. This study determined whether polyamines are required for the stimulation of cell migration by altering K(+) channel gene expression, E(m), and [Ca(2+)](cyt) in intestinal epithelial cells (IEC-6). The specific inhibitor of polyamine synthesis, alpha-difluoromethylornithine (DFMO, 5 mM), depleted cellular polyamines (putrescine, spermidine, and spermine), selectively inhibited Kv1.1 channel (a delayed-rectifier Kv channel) expression, and resulted in membrane depolarization. Because IEC-6 cells did not express voltage-gated Ca(2+) channels, the depolarized E(m) in DFMO-treated cells decreased [Ca(2+)](cyt) as a result of reduced driving force for Ca(2+) influx through capacitative Ca(2+) entry. Migration was reduced by 80% in the polyamine-deficient cells. Exogenous spermidine not only reversed the effects of DFMO on Kv1.1 channel expression, E(m), and [Ca(2+)](cyt) but also restored cell migration to normal. Removal of extracellular Ca(2+) or blockade of Kv channels (by 4-aminopyridine, 1-5 mM) significantly inhibited normal cell migration and prevented the restoration of cell migration by exogenous spermidine in polyamine-deficient cells. These results suggest that polyamine-dependent intestinal epithelial cell migration may be due partially to an increase of Kv1.1 channel expression. The subsequent membrane hyperpolarization raises [Ca(2+)](cyt) by increasing the driving force (the electrochemical gradient) for Ca(2+) influx and thus stimulates cell migration.
Wang
J Y
JY
Department of Surgery, University of Maryland School of Medicine, Baltimore, Maryland 21201, USA. jwang@smail.umaryland.edu
Wang
J
J
Golovina
V A
VA
Li
L
L
Platoshyn
O
O
Yuan
J X
JX
D000818:Animals; D016375:Antisense Elements (Genetics); D002118:Calcium / Q000378:metabolism / Q000493:pharmacokinetics; D015220:Calcium Channels / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects*; D002478:Cells, Cultured; D000518:Eflornithine / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000737:chemistry / Q000166:cytology* / Q000201:enzymology; D015870:Gene Expression / Q000187:drug effects / Q000502:physiology; D007091:Image Processing, Computer-Assisted; D007422:Intestines / Q000166:cytology; D051662:Kv1.1 Potassium Channel; D065108:Ornithine Decarboxylase Inhibitors; D011073:Polyamines / Q000494:pharmacology*; D015221:Potassium Channels / Q000235:genetics* / Q000378:metabolism; D024642:Potassium Channels, Voltage-Gated; D051381:Rats; D013095:Spermidine / Q000494:pharmacology
D016375:Antisense Elements (Genetics); D015220:Calcium Channels; D004791:Enzyme Inhibitors; D065108:Ornithine Decarboxylase Inhibitors; D011073:Polyamines; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D051662:Kv1.1 Potassium Channel; D002118:Calcium; D013095:Spermidine; D000518:Eflornithine