A comparison of the ability of Leu⁸- And pro⁸-oxytocin to regulate intracellular Ca²¹ and Ca²¹-activated K¹ channels at human and marmoset oxytocin receptors ^S

The neurohypophyseal hormone oxytocin (OT) regulates biologic functions in both peripheral tissues and the central nervous system. In the central nervous system, OT influences social processes, including peer relationships, maternal-infant bonding, and affiliative social relationships. In mammals, the nonapeptide OT structure is highly conserved with leucine in the eighth position (Leu⁸-OT). In marmosets (Callithrix), a nonsynon-ymous nucleotide substitution in the OXT gene codes for proline in the eighth residue position (Pro⁸-OT). OT binds to its cognate G protein–coupled receptor (OTR) and exerts diverse effects, including stimulation (G_s) or inhibition (G_i/o) of adenylyl cyclase, stimulation of potassium channel currents (G_i), and activation of phospholipase C (G_q). Chinese hamster ovary cells expressing marmoset or human oxytocin receptors (mOTRs or hOTRs, respectively) were used to characterize OT signaling. At the mOTR, Pro⁸-OT was more efficacious than Leu⁸-OT in measures of G_q activation, with both peptides displaying subnanomolar potencies. At the hOTR, neither the potency nor efficacy of Pro⁸-OT and Leu⁸-OT differed with respect to G_q signaling. In both mOTR- and hOTR-expressing cells, Leu⁸-OT was more potent and modestly more efficacious than Pro⁸-OT in inducing hyperpolarization. In mOTR cells, Leu⁸-OT–induced hyperpolarization was modestly inhibited by pretreatment with pertussis toxin (PTX), consistent with a minor role for G_i/o activation; however, the Pro⁸-OT response in mOTR and hOTR cells was PTX insensitive. These findings are consistent with membrane hyperpolarization being largely mediated by a G_q signaling mechanism leading to Ca²¹-dependent activation of K¹ channels. Evaluation of the influence of apamin, charybdotoxin, paxilline, and TRAM-34 demonstrated involvement of both intermediate and large conductance Ca²¹-activated K¹ channels.