Fecal Microbiota Transfer in Sleep-Deprived Mice

Modified according to the previous literature [16 (link),17 (link)], the feces of sleep-deprived mice were collected once every other day, all feces were pooled together, and then homogenized at 100 mg/mL and supplemented with saline (Solarbio Science & Technology Co., Ltd., Beijing, China). The homogenized solution was centrifuged at 500 rpm for 10 min, and the supernatant was the bacterial suspension. An amount of 100 μL/10 g of the bacterial suspension was transplanted into normal sleeping mice via oral gavage three times a week for 6 weeks. Before FMT, recipient mice were treated with 200 μL of an antibiotic mixture (ampicillin (Shanghai Macklin Biochemical Co., Ltd., Shanghai, China), 1 g/L; metronidazole (Shanghai Macklin Biochemical Co., Ltd., Shanghai, China), 1 g/L; neomycin (Shanghai Macklin Biochemical Co., Ltd., Shanghai, China), 1 g/L; vancomycin (Shanghai Macklin Biochemical Co., Ltd., Shanghai, China), 0.5 g/L) three times, one day apart. FMT was stopped two days before dissection. Fecal samples from the donor and recipient were collected in a sample stabilizer and cryopreserved for subsequent gut microbiota analysis.