Co-expression of the 5-HT 3B Serotonin Receptor Subunit Alters the Biophysics of the 5-HT 3 Receptor

Abstract

Homomeric complexes of 5-HT 3A receptor subunits form a ligand-gated ion channel. This assembly does not fully reproduce the biophysical and pharmacological properties of native 5-HT 3 receptors which might contain the recently cloned 5-HT 3B receptor subunit. In the present study, heteromeric assemblies containing human 5-HT 3A and 5-HT 3B subunits were expressed in HEK 293 cells to detail the functional diversity of 5-HT 3 receptors. We designed patch-clamp experiments with homomeric (5-HT 3A) and heteromeric (5-HT 3AB) receptors to emphasize the kinetics of channel activation and desensitization. Co-expression of the 5-HT 3B receptor subunit reduced the sensitivity for 5-HT (5-HT 3A receptor: EC 50 3 μM, Hill coefficient 1.8; 5-HT 3AB receptor: EC 50 25 μM, Hill coefficient 0.9) and markedly altered receptor desensitization. Kinetic modeling suggested that homomeric receptors, but not heteromeric receptors, desensitize via an agonist-induced open-channel block. Furthermore, heteromeric 5-HT 3AB receptor assemblies recovered much faster from desensitization than homomeric 5-HT 3A receptor assemblies. Unexpectedly, the specific 5-HT 3 receptor agonist mCPBG induced an open-channel block at both homomeric and heteromeric receptors. Because receptor desensitization and resensitization massively affect amplitude, duration, and frequency of synaptic signaling, these findings are evidence in favor of a pivotal role of subunit composition of 5-HT 3 receptors in serotonergic transmission.