Subtle Differences among 5-HT₃AC, 5-HT₃AD, and 5-HT₃AE Receptors Are Revealed by Partial Agonists

5-HT₃ receptors are members of the Cys-loop family of ligand-gated ion channels, and, like most members of this family, there are multiple subunits that can contribute to functional pentameric receptors. 5-HT₃A and 5-HT₃AB receptors have been extensively characterized, but there are few studies on 5-HT₃AC, 5-HT₃AD, and 5-HT₃AE receptors. Here we explore the properties of a range of partial agonists at 5-HT₃AC, 5-HT₃AD, and 5-HT₃AE receptors following expression in Xenopus oocytes. The data show that the characteristics of receptor activation differ in the different heteromeric receptors when they are challenged with 5-HT, m-chlorophenylbiguanide (mCPBG), varenicline, 5-fluorotryptamine (5-FT), or thymol. 5-HT, 5-FT, varenicline, and mCPBG activation of 5-HT₃AC, 5-HT₃AD, and 5-HT₃AE receptors yields similar EC₅₀s to homomeric 5-HT₃A receptors, but maximal responses differ. There are also differences in the levels of potentiation by thymol, which is greater at 5-HT₃A receptors than 5-HT₃AB, 5-HT₃AC, 5-HT₃AD, or 5-HT₃AE receptors. Docking thymol into the receptor indicates a different residue in the transmembrane domain could provide an explanation for these data. Overall our study suggests that 5-HT₃AC, 5-HT₃AD, and 5-HT₃AE have distinct pharmacological profiles to those of 5-HT₃A and 5-HT₃AB receptors; this is likely related to their distinct roles in the nervous system, consistent with their differential association with various disorders. Thus, these data pave the way for drugs that can specifically target these proteins.