Expression of voltage-gated calcium channel subunit alpha1C in epidermal keratinocytes and effects of agonist and antagonists of the channel on skin barrier homeostasis.
Recent studies demonstrated that skin surface electric conditions affect epidermal permeability barrier homeostasis. These results suggest the existence of voltage sensor on the keratinocytes of the epidermis. On the contrary, specific blockers of the voltage-gated calcium channel (VGCC) also affect epidermal barrier homeostasis, but the existence and function of the channel has not been determined. We demonstrated here immunohistochemically the expression of the main subunit of the L-type VGCC, alpha1C, which alone has a calcium channel function, in mouse and human epidermis. Immunostaining, RT-PCR, and Western blotting were carried out to detect the channel protein. Messenger RNA of alpha1C was also detected in mouse epidermis and human keratinocyte culture by RT-PCR. We also evaluated the function of the channel in the cultured human keratinocytes. Previously, we demonstrated that influx of calcium ion into epidermal keratinocytes delayed the barrier recovery after barrier disruption and topical application of calcium channel blocker accelerated the barrier recovery. In this study, topical application of nifedipine and R-(+)-BAY K8644 after tape stripping of hairless mice accelerated the barrier repair rate while application of S-(-)-BAY K8644 delayed the barrier recovery. These results suggest that the VGCC exists on epidermal keratinocytes and plays an important role in skin barrier homeostasis.
Denda
Mitsuhiro
M
Shiseido Research Center, Kanazawa-ku, Yokohama, Japan. mitsuhiro.denda@to.shiseido.c.jp
Fujiwara
Shigeyoshi
S
Hibino
Toshihiko
T
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000494:pharmacology; D000818:Animals; D002120:Calcium Channel Agonists / Q000494:pharmacology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020746:Calcium Channels, L-Type / Q000235:genetics* / Q000378:metabolism; D002478:Cells, Cultured; D000078404:Epidermal Cells; D004817:Epidermis / Q000187:drug effects / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006706:Homeostasis / Q000187:drug effects; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D051379:Mice; D008812:Mice, Hairless; D009543:Nifedipine / Q000494:pharmacology; D010539:Permeability / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014700:Verapamil / Q000494:pharmacology
D002120:Calcium Channel Agonists; D002121:Calcium Channel Blockers; D020746:Calcium Channels, L-Type; C419385:L-type calcium channel alpha(1C); D012333:RNA, Messenger; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D014700:Verapamil; D009543:Nifedipine