Expression of thermosensitive two-pore domain K + channels in human keratinocytes cell line HaCaT cells

Dawon Kang, Sung Hee Kim, Eun Mi Hwang, Oh Sang Kwon, Hae Young Yang, Eun Sook Kim, Tae Hyun Choi, Jae-Yong Park, Seong Geun Hong, Jaehee Han

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Recent studies have shown that keratinocytes can sense temperature via thermo-transient receptor potential (TRP) channels. It is not known whether other thermosensitive ion channels such as TREK-1, TREK-2 and TRAAK (TREKs/TRAAK) that are members of the two-pore domain K + (K 2P ) channel family are expressed in human keratinocytes. Here, we identified the expression of TREKs/TRAAK in human keratinocytes-derived cell line HaCaT cells using RT-PCR, immunocytochemistry, Western blot analysis and patch-clamp technique. RT-PCR showed that all six K 2P channels tested (TASK-1, TASK-3, TREK-1, TREK-2, TRAAK and TASK-2) were expressed in HaCaT cells, as well as in skin and dorsal root ganglion (DRG) of rat. The expression of TREKs/TRAAK mRNA identified by RT-PCR was further studied at the protein level. Using anti-TREK-1, -TREK-2 and -TRAAK, bands of ∼46, ∼60 and ∼43 kDa, respectively, were observed at plasma membrane of HaCaT cells. Immunostaining also showed that TREK-1, TREK-2 and TRAAK were expressed in all area of cells including plasma membrane. Whole-cell K + currents recorded from HaCaT cells were activated by arachidonic acid and heat. These results suggest that TREKs/TRAAK channels could act as thermosensors in human keratinocytes.

Original languageEnglish
Pages (from-to)1016-1022
Number of pages7
JournalExperimental Dermatology
Volume16
Issue number12
DOIs
Publication statusPublished - 2007 Dec 1
Externally publishedYes

Fingerprint

Keratinocytes
Cell membranes
Cells
Cell Line
Transient Receptor Potential Channels
Polymerase Chain Reaction
Thermosensing
Clamping devices
Ion Channels
Cell Membrane
Arachidonic Acid
Rats
Skin
Spinal Ganglia
Patch-Clamp Techniques
Messenger RNA
potassium channel protein TREK-1
Hot Temperature
Western Blotting
Immunohistochemistry

Keywords

  • Keratinocytes
  • Potassium channels
  • Tandem-pore domain
  • Temperature sense

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Dermatology

Cite this

Expression of thermosensitive two-pore domain K + channels in human keratinocytes cell line HaCaT cells . / Kang, Dawon; Kim, Sung Hee; Hwang, Eun Mi; Kwon, Oh Sang; Yang, Hae Young; Kim, Eun Sook; Choi, Tae Hyun; Park, Jae-Yong; Hong, Seong Geun; Han, Jaehee.

In: Experimental Dermatology, Vol. 16, No. 12, 01.12.2007, p. 1016-1022.

Research output: Contribution to journalArticle

Kang, D, Kim, SH, Hwang, EM, Kwon, OS, Yang, HY, Kim, ES, Choi, TH, Park, J-Y, Hong, SG & Han, J 2007, ' Expression of thermosensitive two-pore domain K + channels in human keratinocytes cell line HaCaT cells ', Experimental Dermatology, vol. 16, no. 12, pp. 1016-1022. https://doi.org/10.1111/j.1600-0625.2007.00626.x
Kang, Dawon ; Kim, Sung Hee ; Hwang, Eun Mi ; Kwon, Oh Sang ; Yang, Hae Young ; Kim, Eun Sook ; Choi, Tae Hyun ; Park, Jae-Yong ; Hong, Seong Geun ; Han, Jaehee. / Expression of thermosensitive two-pore domain K + channels in human keratinocytes cell line HaCaT cells In: Experimental Dermatology. 2007 ; Vol. 16, No. 12. pp. 1016-1022.
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AU - Kang, Dawon

AU - Kim, Sung Hee

AU - Hwang, Eun Mi

AU - Kwon, Oh Sang

AU - Yang, Hae Young

AU - Kim, Eun Sook

AU - Choi, Tae Hyun

AU - Park, Jae-Yong

AU - Hong, Seong Geun

AU - Han, Jaehee

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AB - Recent studies have shown that keratinocytes can sense temperature via thermo-transient receptor potential (TRP) channels. It is not known whether other thermosensitive ion channels such as TREK-1, TREK-2 and TRAAK (TREKs/TRAAK) that are members of the two-pore domain K + (K 2P ) channel family are expressed in human keratinocytes. Here, we identified the expression of TREKs/TRAAK in human keratinocytes-derived cell line HaCaT cells using RT-PCR, immunocytochemistry, Western blot analysis and patch-clamp technique. RT-PCR showed that all six K 2P channels tested (TASK-1, TASK-3, TREK-1, TREK-2, TRAAK and TASK-2) were expressed in HaCaT cells, as well as in skin and dorsal root ganglion (DRG) of rat. The expression of TREKs/TRAAK mRNA identified by RT-PCR was further studied at the protein level. Using anti-TREK-1, -TREK-2 and -TRAAK, bands of ∼46, ∼60 and ∼43 kDa, respectively, were observed at plasma membrane of HaCaT cells. Immunostaining also showed that TREK-1, TREK-2 and TRAAK were expressed in all area of cells including plasma membrane. Whole-cell K + currents recorded from HaCaT cells were activated by arachidonic acid and heat. These results suggest that TREKs/TRAAK channels could act as thermosensors in human keratinocytes.

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KW - Tandem-pore domain

KW - Temperature sense

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