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This version was published on December 1, 2005
Journal of Biomolecular Screening, Vol. 10, No. 8, 841-848 (2005)
DOI: 10.1177/1087057105280285

Ecdysone-Based System for Controlled Inducible Expression of Metabotropic Glutamate Receptor Subtypes 2, 5, and 8

Patrick. M. Downey

schering-plough Research Institute, Sun Raffaele Biomedical Science park, Via Olgettina 58, 20132 Milano, Italypatrick.downey{at}spcorp.com

Gianluca Lozza

Roberta Petrò

Enrica Diodato

Chiara Foglia

Federica Bottazzoli

Rossella Brusa

Tatiana Asquini

Angelo Reggiani

Mariagrazia Grilli

Stable and inducible expression of human metabotropic glutamate receptor types 2, 5, and 8 was achieved in HEK293 cells using the ecdysone inducible system. Treatment of the respective cell lines with ponasterone A resulted in time and concentration-dependent induction of receptor expression. In all cases, the functional activation of receptorswas determined bymeasuring increases in intracellular calcium. The physiologically G• i-coupled receptors mGluR2 andm GluR8 were successfully coupled to phospholipase Cactivation using the chimeric Gprotein G• q/o. The pharmacological properties of recombinant receptors were characterized and proved to be similar to native receptors. Our data suggest that the ecdysone system has a number of characteristics thatmake itwell suited for expressing mGluRs and that the combined use of this systemand chimeric G proteins allows receptors to be characterized using a rapid and straightforward Ca 2+assay.

Key Words: metabotropic glutamate receptors • inducible expression • chimeric G proteins


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