title: Metamorphosis inhibition: an alternative rearing protocol for the newt, Cynops pyrrhogaster.
abstract: The newt is an indispensable model animal, of particular utility for regeneration studies. Recently, a high-throughput transgenic protocol was established for the Japanese common newt, Cynops pyrrhogaster. For studies of regeneration, metamorphosed animals may be favorable; however, for this species, there is no efficient protocol for maintaining juveniles after metamorphosis in the laboratory. In these animals, survival drops drastically after metamorphosis as their foraging behaviour changes to adapt to a terrestrial habitat, making feeding in the laboratory with live or moving foods more difficult. To elevate the efficiency of laboratory rearing of this species, we examined metamorphosis inhibition (Ml) protocols to bypass the period (four months to two years after hatching) in which the animal feeds exclusively on moving foods. We found that approximately 30% of animals survived after 2-year Ml, and that the survivors continuously grew, only with static food while maintaining their larval form and foraging behaviour in 0.02% thiourea (TU) aqueous solution, then metamorphosed when returned to a standard rearing solution even after 2-year-MI. The morphology and foraging behavior (feeding on static foods in water) of these metamorphosed newts resembled that of normally developed adult newts. Furthermore, they were able to fully regenerate amputated limbs, suggesting regenerative capacity is preserved in these animals. Thus, controlling metamorphosis with TU allows newts to be reared with the same static food under aqueous conditions, providing an alternative rearing protocol that offers the advantage of bypassing the critical period and obtaining animals that have grown sufficiently for use in regeneration studies.
authors.affiliation: Faculty of Life and Environmental Sciences, University of Tsukuba, lbaraki 305-8572, Japan. chichiba@biol.tsukuba.ac.jp
authors.forename: Chikafumi
authors.initials: C
authors.lastname: Chiba
authors.forename: Shouta
authors.initials: S
authors.lastname: Yamada
authors.forename: Hibiki
authors.initials: H
authors.lastname: Tanaka
authors.forename: Maiko
authors.initials: M
authors.lastname: Inae-Chiba
authors.forename: Tomoya
authors.initials: T
authors.lastname: Miura
authors.forename: Martin Miguel
authors.initials: MM
authors.lastname: Casco-Robles
authors.forename: Taro
authors.initials: T
authors.lastname: Yoshikawa
authors.forename: Wataru
authors.initials: W
authors.lastname: Inami
authors.forename: Aki
authors.initials: A
authors.lastname: Mizuno
authors.forename: Md Rafiqul
authors.initials: MR
authors.lastname: Islam
authors.forename: Wenje
authors.initials: W
authors.lastname: Han
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Yasumuro
authors.forename: Mikiko
authors.initials: M
authors.lastname: Matsumoto
authors.forename: Miyako
authors.initials: M
authors.lastname: Takayanagi
mesh_terms: D000818:Animals; D007756:Laboratory Animal Science; D008675:Metamorphosis, Biological / Q000187:drug effects* / Q000502:physiology*; D012455:Salamandridae / Q000254:growth & development*; D013890:Thiourea / Q000494:pharmacology*
chemical_list: D013890:Thiourea