Enhancement of Ca(2+) influx and ciliary beating by membrane hyperpolarization due to ATP-sensitive K(+) channel opening in mouse airway epithelial cells.
Among the several types of cells composing the airway epithelium, the ciliary cells are responsible for one of the most important defense mechanisms of the airway epithelium: the transport of inhaled particles back up into the throat by coordinated ciliary movement. Changes in the cytoplasmic Ca(2+) concentration ([Ca(2+)]i) are the main driving force controlling the ciliary activity. In mouse ciliary cells, membrane hyperpolarization from -20 to -60 mV under whole-cell voltage-clamp induced a slow but significant [Ca(2+)]i rise in a reversible manner. This rise was completely inhibited by the removal of Ca(2+) from the extracellular solution. Application of diazoxide, an ATP-dependent K(+) channel opener, dose-dependently induced a membrane hyperpolarization (EC50 = 2.3 μM), which was prevented by the addition of 5 μM glibenclamide. An inwardly rectifying current was elicited by the application of 10 μM diazoxide and suppressed by subsequent addition of 5 μM glibenclamide. Moreover, the application of 10 μM diazoxide induced a significant [Ca(2+)]i rise and facilitated ciliary movement. Multi-cell reverse-transcription polymerase chain reaction analyses and immunocytochemical staining suggested that the subunit combination of Kir6.2/SUR2B and possibly also Kir6.1/SUR2B is expressed in ciliary cells. The confocal Ca(2+) imaging analyses suggested that the [Ca(2+)]i rise induced by diazoxide occurred preferentially in the apical submembrane region. In conclusion, the application of a KATP channel opener to airway ciliary cells induces membrane hyperpolarization and thereby induces a [Ca(2+)]i rise via the facilitation of Ca(2+) influx through the non-voltage-dependent Ca(2+) permeable channels. Therefore, a KATP opener may be beneficial in facilitating ciliary movement.
Department of Molecular and Cellular Pharmacology, Graduate School of Pharmaceutical Sciences (T.O., E.S., Y.S., H.Y., Y.I.) and Nanomaterial Toxicology Project (H.T.), Nagoya City University, Nagoya, Japan; and Department of Pharmacology, Division of Pathological Sciences, Kyoto Pharmaceutical University, Kyoto, Japan (S.O.).
Teruya
T
Ohba
Eiji
E
Sawada
Yoshiaki
Y
Suzuki
Hisao
H
Yamamura
Susumu
S
Ohya
Hiroyuki
H
Tsuda
Yuji
Y
Imaizumi
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002923:Cilia / Q000187:drug effects / Q000378:metabolism; D003981:Diazoxide / Q000494:pharmacology; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology; D054086:KATP Channels / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D024661:Potassium Channels, Inwardly Rectifying / Q000378:metabolism*; D020545:Respiratory Mucosa / Q000187:drug effects / Q000378:metabolism*; D064233:Sulfonylurea Receptors / Q000378:metabolism*; D000099161:Kcnj11 Channel
D002118:Calcium; D003981:Diazoxide; D054086:KATP Channels; D024661:Potassium Channels, Inwardly Rectifying; D064233:Sulfonylurea Receptors; D000099161:Kcnj11 Channel; C577511:Abcc9 protein, mouse