Gekko japonicus breeding and modeling

Adult G. japonicus with intact tails were grouped into control and experimental groups after health evaluation by veterinarians. The number of males and females were equal in each group. The average body weight was 4.4 ± 0.495 g. All geckos had free access to mealworms and sterilized water during the entire experiment. The culture incubator was set at 28 °C, 17 °C or 10 °C based on the test protocol. All animal experimental protocols were approved by the Institutional Animal Care and Use Committee of the Nantong University (Anima protocol: S20190405-402).

Tail autotomy was exerted following the protocols issued previously by our lab members²⁴. Concisely, caudectomy was performed by locating a nylon slipknot at the 6th tail segment and then pulling gently to simulate the stimulation of autotomy in the natural environment. The successful regeneration was determined by the featured viewing as scab shedding and exposing of pink epithelium at ~7 day. The delayed scab shedding indicated deficiency of the wounding epithelium formation, which always led to failure of regeneration. The subsequent regeneration was evaluated by the length of regenerated tails that described in our previous study²⁴, mainly detected by morphology observation, pathological investigation and molecular markers indication that detailed below.