10.1021/acschemneuro.0c00199.s001 Cassandra J. Hatzipantelis Cassandra J. Hatzipantelis Yao Lu Yao Lu Daisy L. Spark Daisy L. Spark Christopher J. Langmead Christopher J. Langmead Gregory D. Stewart Gregory D. Stewart β‑Arrestin-2-Dependent Mechanism of GPR52 Signaling in Frontal Cortical Neurons American Chemical Society 2020 BTBZ transfected HEK 293T cells GPR 52 activation GPR 52 Signaling β- arrestin -2 HEK 293T cells GPCR GPR 52 agonist neuron Frontal Cortical Neurons cAMP accumulation β- arrestin -2-dependent mechanism GPR 52 agonists CREB phosphorylation ERK 3- G αs receptor GPR 52 2020-07-07 13:42:22 Journal contribution https://acs.figshare.com/articles/journal_contribution/_Arrestin-2-Dependent_Mechanism_of_GPR52_Signaling_in_Frontal_Cortical_Neurons/12619866 The orphan Gαs-coupled receptor GPR52 is expressed exclusively in the brain, predominantly in circuitry relating to symptoms of neuropsychiatric and cognitive disorders such as schizophrenia. While GPR52 agonists have displayed antipsychotic and procognitive efficacy in murine models, there remains limited evidence delineating the molecular mechanisms of these effects. Indeed, previous studies have solely reported canonical cAMP signaling and CREB phosphorylation downstream of GPR52 activation. In the present study, we demonstrated that the synthetic GPR52 agonist, 3-BTBZ, equipotently induces cAMP accumulation, ERK1/2 phosphorylation, and β-arrestin-1 and -2 recruitment in transfected HEK293T cells. In cultured frontal cortical neurons, however, 3-BTBZ-induced ERK1/2 phosphorylation was significantly more potent than cAMP signaling, with a more prolonged signaling profile than that in HEK293T cells. Furthermore, knock down of β-arrestin-2 in frontal cortical neurons abolished 3-BTBZ-induced ERK1/2 phosphorylation, but not cAMP accumulation. These results suggest a β-arrestin-2-dependent mechanism for GPR52-mediated ERK1/2 signaling, which may link to cognitive function <i>in vivo</i>. Finally, these findings highlight the context-dependence of GPCR signaling in recombinant cells and neurons, offering new insights into translationally relevant GPR52 signaling mechanisms.