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

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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

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Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Dermatology

Cite this

Kang, D., Kim, S. H., Hwang, E. M., Kwon, O. S., Yang, H. Y., Kim, E. S., Choi, T. H., Park, J-Y., Hong, S. G., & Han, J. (2007). Expression of thermosensitive two-pore domain K + channels in human keratinocytes cell line HaCaT cells Experimental Dermatology, 16(12), 1016-1022. https://doi.org/10.1111/j.1600-0625.2007.00626.x