Expression of thyrotropin-releasing hormone receptors by adenovirus-mediated gene transfer reveals that thyrotropin-releasing hormone desensitization is cell specific.
Biological studies of seven-transmembrane region G protein-coupled receptors have been restricted by available techniques for gene transfer into mammalian cells. We have created a highly efficient adenovirus-based expression vector for the thyrotropin-releasing hormone (TRH) receptor (TRH-R), AdCMVmTRHR, to circumvent difficulties encountered when transient or stable plasmid expression systems are used. We show that infection with AdCMVmTRHR results in fully functional TRH-Rs, which can be expressed in a broad range of mammalian cell types, including those resistant to conventional transient transfection. TRH-Rs can be expressed at high levels, up to 2 x 10(6) receptors/cell. Expression in several cell lines in culture reveals that rapid TRH-R desensitization by TRH and phorbol 12-myristate 13-acetate is cell type specific. The versatility of adenovirus-mediated gene transfer and expression of TRH-Rs not only facilitates in vitro studies of TRH-R biology but also provides a valuable in vivo expression vector capable of extending TRH-R studies to animal model systems.
Falck-Pedersen
E
E
Department of Microbiology, Cornell University Medical College, New York, New York 10021.
Heinflink
M
M
Alvira
M
M
Nussenzveig
D R
DR
Gershengorn
M C
MC
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015536:Down-Regulation / Q000187:drug effects; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D066298:In Vitro Techniques; D051379:Mice; D018025:Receptors, Thyrotropin-Releasing Hormone / Q000235:genetics / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013973:Thyrotropin-Releasing Hormone / Q000494:pharmacology
D018025:Receptors, Thyrotropin-Releasing Hormone; D013973:Thyrotropin-Releasing Hormone; D013755:Tetradecanoylphorbol Acetate