Pharmacological profiling of native group II metabotropic glutamate receptors in primary cortical neuronal cultures using a FLIPR

► We developed a high-throughput native group II mGluR functional assay using FLIPR. ► mGluR group II agonists inhibit Ca2+ oscillation frequency in primary neurones. ► mGlu2 receptor agonist and PAMs inhibit Ca2+ oscillation frequency in primary neurones. ► mGlu2 and/or mGlu3 receptor deletion revealed that LY541850 is a partial mGlu2 agonist and an mGlu3 receptor antagonist. ► Our assay can discern between mGlu2 and mGlu3 receptor contributions using KO cultures.