Corticotropin-releasing factor regulates the development in the direct developing frog, Eleutherodactylus coqui

Direct developing frogs lack a free-living larval phase, such that miniature adults hatch directly from the eggs. Even under such extreme reorganization of the ancestral biphasic developmental pattern, direct developers still undergo thyroid hormone (TH)-dependent post-embryonic development. Hypothalamic regulation of TH synthesis and release plays a central role in controlling the timing of metamorphosis in biphasic developers. In particular, the neuropeptide corticotropin-releasing factor (CRF) regulates TH in tadpoles, but in adults, both thyrotropin-releasing hormone (TRH) and CRF regulate TH. Because direct developers lack a tadpole stage, it was not clear whether hypothalamic regulation of TH would be tadpole-like or adult-like prior to hatching. To test this, we injected pre-hatching Eleutherodactylus coqui daily with CRF, TRH or astressin (a CRF receptor blocker). CRF but not TRH significantly accelerated the developmental rate compared to controls. Astressin-treated animals showed a near complete developmental arrest, which confirmed that development requires CRF. To support the idea that CRF acts to regulate development in E. coqui via thyroid physiology, we showed the TH-direct response gene TRβ is up-regulated 24 and 48 h after CRF injection. In addition, treatment with 50 nM T3 (triiodothyronine, the active form of TH) increased the developmental rate similar to CRF injections. Our results extend the evidence for a cryptic metamorphosis in direct developers by showing that neuroendocrine signaling is conserved between biphasic and direct developers. Furthermore, the conserved neuroendocrine regulation implies that changes at the peripheral level of hormone action underlie the evolution of the radically divergent development in direct developers.