Construction of a stable cell line uniformly expressing the rat TRPV1 receptor.
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.
Sándor
Zoltán
Z
Department of Pharmacology and Pharmacotherapy, Faculty of Medicine, Neuropharmacology Research Group of the Hungarian Academy of Sciences, University of Pécs, Szigeti u. 12, H-7624 Pécs, Hungary.
Varga
Angelika
A
Horváth
Péter
P
Nagy
Barbara
B
Szolcsányi
János
J
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002211:Capsaicin / Q000494:pharmacology; D002460:Cell Line; D002478:Cells, Cultured; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D016257:Fura-2; D005727:Ganglia, Spinal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D051381:Rats; D050916:TRPV Cation Channels / Q000378:metabolism*
D005456:Fluorescent Dyes; D050916:TRPV Cation Channels; C494484:TRPV1 receptor; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D002211:Capsaicin; D002118:Calcium; D016257:Fura-2