Construction of a stable cell line uniformly expressing the rat TRPV1 receptor

Zoltán Sándor, Angelika Varga, Péter Horváth, Barbara Nagy, János Szolcsányi

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

We constructed and analyzed a new cell line called HT5-1, which stably expresses an enhanced green fluorescent protein-tagged version of the rat vanilloid receptor 1 (VR1/TRPV1). The fluorescent receptor allowed easy measurement of receptor expression and expression level-based purification of cells via fluorescence-activated cell sorting. The HT5-1 cell line was compared to cells transiently transfected with the fluorescent receptor, to cells expressing the native rat vanilloid receptor, and to isolated capsaicin-sensitive rat trigeminal sensory neurons. Fura-2 microfluorimetry measurements of the calcium influx upon capsaicin induction showed that, by contrast to transiently transfected cells, HT5-1 cells respond uniformly to the stimulation, due to the similar level of receptor expression in individual cells. HT5-1 cells showed similar behaviour to isolated trigeminal root ganglion neurons, including marked tachyphylaxis upon repeated capsaicin induction, and a lack of calcium ion release from intracellular storage sites.

Original languageEnglish
Pages (from-to)499-514
Number of pages16
JournalCellular and Molecular Biology Letters
Volume10
Issue number3
Publication statusPublished - Jan 1 2005

Keywords

  • Capsaicin Receptor
  • Flow Cytometry
  • Fluorescence-Activated Cell Sorting
  • Green Fluorescent Protein
  • Stable Cell Line
  • TRG Neurons
  • TRPV1 Receptor Expression
  • Tachyphylaxis

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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