Functional Characterization of Thyrostimulin in Amphioxus Suggests an Ancestral Origin of the TH Signaling Pathway

Thyrostimulin, consisting of GpA2 and GpB5 subunits, has been identified in amphioxus, but to date, little is known about the roles of GPA2/GPB5‒type hormone in this evolutionarily important animal. We showed here that amphioxus GpA2, GpB5, and TSH receptor (TSHR) represent the archetypes of vertebrate TSHα, TSHβ, and TSHR, respectively, and both gpa2 and gpb5 were coexpressed in the Hatschek pit, a homolog of the vertebrate pituitary, in amphioxus. We also showed that recombinant amphioxus GpA2 and GpB5, like zebrafish TSHα and TSHβ, bound to both amphioxus and zebrafish TSHR and that tethered amphioxus thyrostimulin activated both protein kinase A and protein kinase C pathways in the cells expressing amphioxus TSHR. Moreover, we demonstrated that recombinant amphioxus thyrostimulin induced the production of thyroid hormone (TH) T4. Because genuine TSH is absent in amphioxus and thyrostimulin is the only and sole glycoprotein hormone, our data likely provide evidence that amphioxus thyrostimulin is a functional glycoprotein hormone that plays a role as TSH does in vertebrates. The data also suggest that the TH signaling pathway evolved in the basal chordate more than 500 million years ago.