Tetracycline-based system for controlled inducible expression of group III metabotropic glutamate receptors.
A stable and inducible expression of metabotropic glutamate receptor type 4, 7, and 8 was obtained in T-REx 293 cells using the tetracycline system. Tetracycline administration to the cell medium resulted in rapid induction and time-dependent expression of mGlu receptors, which also correlates with its functionality in a cAMP accumulation assay. The pharmacological properties of recombinant mGlu receptors were verified using orthosteric and allosteric ligands. Data suggest that the Tet-on inducible system is suitable for functional mGlu receptors' expression and characterization by means of the cAMP accumulation assay. It makes this system a precise, reproducible, and large-scale screening method, as well as a reasonable tool to study signaling properties of mGlu receptors.
Department of Neurobiology, Institute of Pharmacology, Polish Academy of Sciences, Kraków, Smętna, Poland chruscic@if-pan.krakow.pl.
Barbara
B
Chruścicka
Department of Neurobiology, Institute of Pharmacology, Polish Academy of Sciences, Kraków, Smętna, Poland.
Grzegorz
G
Burnat
Department of Neurobiology, Institute of Pharmacology, Polish Academy of Sciences, Kraków, Smętna, Poland.
Piotr
P
Brański
Department of Neurobiology, Institute of Pharmacology, Polish Academy of Sciences, Kraków, Smętna, Poland.
Paulina
P
Chorobik
Department of Neuropsychopharmacology, Institute of Pharmacology, Polish Academy of Sciences, Kraków, Smętna, Poland.
Tomasz
T
Lenda
Department of Neurobiology, Institute of Pharmacology, Polish Academy of Sciences, Kraków, Smętna, Poland.
Marcin
M
Marciniak
Department of Neurobiology, Institute of Pharmacology, Polish Academy of Sciences, Kraków, Smętna, Poland Faculty of Health Sciences, Medical College, Jagiellonian University, Kraków, Grzegórzecka, Poland.
Andrzej
A
Pilc
D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression; D005786:Gene Expression Regulation / Q000187:drug effects*; D018698:Glutamic Acid / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D008024:Ligands; D011401:Promoter Regions, Genetic; D018094:Receptors, Metabotropic Glutamate / Q000235:genetics* / Q000378:metabolism; D013752:Tetracycline / Q000494:pharmacology*; D013997:Time Factors
D008024:Ligands; D018094:Receptors, Metabotropic Glutamate; C102719:metabotropic glutamate receptor 7; C111435:metabotropic glutamate receptor 8; D018698:Glutamic Acid; D000242:Cyclic AMP; D013752:Tetracycline; C108823:metabotropic glutamate receptor 4
HEK293 cell line; cyclic AMP accumulation assay; inducible expression system; metabotropic glutamate receptors; tetracycline-regulated promoter