Tetracycline-Based System for Controlled Inducible Expression of Group III Metabotropic Glutamate Receptors

Author:

Chruścicka Barbara1,Burnat Grzegorz1,Brański Piotr1,Chorobik Paulina1,Lenda Tomasz2,Marciniak Marcin1,Pilc Andrzej13

Affiliation:

1. Department of Neurobiology, Institute of Pharmacology, Polish Academy of Sciences, Kraków, Smętna, Poland

2. Department of Neuropsychopharmacology, Institute of Pharmacology, Polish Academy of Sciences, Kraków, Smętna, Poland

3. Faculty of Health Sciences, Medical College, Jagiellonian University, Kraków, Grzegórzecka, Poland

Abstract

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.

Publisher

Elsevier BV

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