Figure 1
Representative recordings of the bioluminescence from the whole bladders of Per2::Luc mice. Whole bladders from neonate (a) and young adult (b) Per2::Luc mice were analyzed for Per2 promoter-driven luciferase activity. (a) Adult Per2::Luc knock-in mice were anesthetized with chloral hydrate (360 mg kg−1, intraperitoneal), and the whole bladders were retrieved. Then, the bladder outlet was tied using thread, the remaining urine was removed using a syringe, and the bladder was filled with MEM (Gibco) supplemented with the appropriate medium. The whole bladder samples were immobilized in a 35-mm culture dish (Corning, Tewksbury, MA, USA) using cell strainers (SPL, Seoul, Korea), and the bioluminescence was continuously recorded using a KRONOS apparatus (ATTO, Tokyo, Japan). (b) To monitor neonate bladders, bladders were obtained from newborn mice, and the tissues were sliced using a blade. The tissue slices were immobilized in a culture insert and incubated for 1 day after a medium change. Then, the tissue slice were synchronized by dexamethasone shock for 3 h. Monitoring was performed using the same method as for the adult bladder tissues. The bioluminescence was continuously recorded for more than 5 days using a KRONOS apparatus as described in Methods.
