Premium
Comparison of TRPV1 on kidney specific sensory neurons and HEK 293 cells
Author(s) -
Wang Hui,
Sui Dexin,
Wang Donna H.,
Garavito Michael R.,
Galligan James J.
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.937.1
Subject(s) - hek 293 cells , trpv1 , transfection , microbiology and biotechnology , chemistry , receptor , transient receptor potential channel , recombinant dna , biology , biochemistry , gene
The transient receptor potential vanilloid 1 (TRPV1) receptor is a ligand‐gated cation channel primarily in sensory nerves. We are interested in the molecular properties of TRPV1 that contributes to salt regulation in the kidney. Therefore, we compared the properties of TRPV1 in kidney specific sensory neurons with several forms of recombinant TRPV1 expressed in human embryonic kidney (HEK)‐293 cells. Fast Blue (FB) was used to retrogradely label sensory neurons innervating the kidney in rats. Three types of TRPV1 plasmids were studied in transiently transfected HEK‐293 cells. Plasmid 1 had a hexa‐His‐tag at the end of the C‐terminus of human TRPV1. Plasmid 2 had an octa‐His‐tag on the N‐terminus and a hexa‐His‐tag on the C‐terminus. Plasmid 3 encoded two copies of the human TRPV1, both with hexa‐His‐tags on the C‐terminus. Immunocytochemistry confirmed TRPV1 expression by FB‐labeled neurons and HEK 293 cells. Whole‐cell recordings done in FB‐labeled neurons and HEK‐293 cells in culture showed that capsaicin (CAP) induced comparable inward currents in neurons and in HEK‐293 cells transfected with plasmids 1 and 2 but not 3. CAP‐induced currents desensitized in FB‐labeled neurons but not in HEK‐293 cells. These data suggest that His tags on heterologously expressed TRPV1 alter receptor desensitization, as the mechanisms regulating TRPV1 desensitization in sensory neurons are present in HEK‐293 cells.