Location and Cell-Type-Specific Bias of Metabotropic Glutamate Receptor, mGlu₅, Negative Allosteric Modulators

Emerging data indicate that G-protein coupled receptor (GPCR) signaling is determined by not only the agonist and a given receptor but also a variety of cell-type-specific factors that can influence a receptor's response. For example, the metabotropic glutamate receptor, mGlu₅, which is implicated in a number of neuropsychiatric disorders such as depression, anxiety, and autism, also signals from inside the cell which leads to sustained Ca²⁺ mobilization versus rapid transient responses. Because mGlu₅ is an important drug target, many negative allosteric modulators (NAMs) have been generated to modulate its activity. Here we show that NAMs such as AFQ056, AZD2066, and RG7090 elicit very different end points when tested in postnatal neuronal cultures expressing endogenous mGlu₅ receptors. For example, AFQ056 fails to block intracellular mGlu₅-mediated Ca²⁺ increases whereas RG7090 is very effective. These differences are not due to differential receptor levels, since about the same number of mGlu₅ receptors are present on neurons from the cortex, hippocampus, and striatum based on pharmacological, biochemical, and molecular data. Moreover, biotinylation studies reveal that more than 90% of the receptor is intracellular in these neurons. Taken together, these data indicate that the tested NAMs exhibit both location-dependent and cell type specific bias for mGlu₅-mediated Ca²⁺ mobilization which may affect clinical outcomes.