Cloning and expression of Ca2+-activated chloride channel from rat brain

Min Jeong Sang, Hye Kyung Park, In Soo Yoon, Jun Ho Lee, Jong Hoon Kim, Choon Gon Jang, Changjoon Lee, Seung Yeol Nah

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

To clone the gene product responsible for the calcium-activated chloride channel (CLCA) in rat brain cerebrum, we performed a reverse transcription-PCR (RT-PCR) with gene-specific primers of a rat EST clone. We successfully cloned a rat brain CLCA (rbCLCA). The full-length cDNA is 2895 bp long and codes for a 902 amino acid protein. The clone consists of four transmembrane domains and shows a 79.1% of significant homology with previously reported mouse smooth muscle chloride channel sequence. We also performed RT-PCR using single neuron and glia, and various tissues to determine the tissue expression of rbCLCA. We found that rbCLCA was expressed in both neuron and glia. In peripheral organs, rbCLCA showed the predominant expressions in cerebrum, cerebellum, kidney, small intestine, and stomach but not in heart, large intestine, liver, lung, and spleen. Whole-cell patch clamp studies in HEK293 cells transfected with the clone identified a niflumic acid (a CLCA channel blocker)-sensitive and voltage-dependent chloride current but we could not observe this chloride current in mock-transfected cells. The identification of genes belonging to the CLCA family from rat brain and its functional expression will help to evaluate its physiological role in brain as anion channel.

Original languageEnglish
Pages (from-to)569-576
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume334
Issue number2
DOIs
Publication statusPublished - 2005 Aug 26
Externally publishedYes

    Fingerprint

Keywords

  • Ca
  • Ca-activated chloride channel
  • Central nervous system
  • Chloride channel
  • Glia
  • Neuron

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Sang, M. J., Park, H. K., Yoon, I. S., Lee, J. H., Kim, J. H., Jang, C. G., Lee, C., & Nah, S. Y. (2005). Cloning and expression of Ca2+-activated chloride channel from rat brain. Biochemical and Biophysical Research Communications, 334(2), 569-576. https://doi.org/10.1016/j.bbrc.2005.06.122