Cane Toad Skin ExtractInduced Upregulation and Increased Interaction of Serotonin 2A and D2 Receptors via Gq/11 Signaling Pathway in CLU213 Cells

Recent evidences show that activation of serotonin 2A receptors (5-HT2AR) by agonists is significant in improving therapeutic activity of disease conditions, such as obsessive-compulsive disorder (OCD). Though the exact molecular mechanism is still not well understood, it is thought to involve agonist-driven, enhanced expression of 5-HT2AR in certain areas of brain, such as the pre-frontal cortex (PFC). Several other reports have also demonstrated association of OCD with lower dopamine receptor (D2R) availability, primarily in the striatum of the brain along with dysfunction of 5-HT2AR-D2R heteromer regulation. We thus hypothesized that compound(s) interacting with this molecular mechanism could be developed as drugs for long-term beneficial effects against OCD. In the present study, we have obtained experimental evidence in cultured neuronal cells (CLU213) that aqueous extract (AE, 50g/mL, P<0.05) of the Australian cane toad skin significantly increased the levels of 5-HT2AR and D2R protein and mRNA expression. AE was also found to enhance the interaction between 5-HT2AR and D2R and formation of expression of 5-HT2AR-D2R heteromer using co-immunoprecipitation and Western blot. Further investigation showed the involvement of classical signaling pathway (G(q/11)-PLC) along with c-FOS transcription factor preferentially in 5-HT2A-mediated agonist activation. These results obtained demonstrated that AE upregulates 5-HT2AR by a mechanism that appears to involve G(q/11)-PLC signaling pathway and c-FOS transcription factor activation. We indicate this enhanced 5-HT2AR and D2R expression and their interaction to induce increased 5-HT2AR-D2R heteromer formation by exposure to AE might provide a molecular mechanism to develop potential novel drug candidates to ameliorate OCD symptoms. J. Cell. Biochem. 118: 979-993, 2017. (c) 2016 Wiley Periodicals, Inc.