Gut microbe: P90
Male Sprague Dawley rats (F0 generation) were exposed to 2.1% sevoflurane for 3 hours on postnatal days 56, 58, and 60 (F0M_S group). F0 males in the F0M_BS and F0M_RS groups received bumetanide or RU486, respectively, prior to each sevoflurane exposure. These males were mated on postnatal day 90 to produce offspring (F1 generation). Gut microbiota were profiled using 16S rRNA gene sequencing. The distal colon of rats for each group were aseptically dissected and the intestinal contents were collected in DNA/RNA Shield solution (Zymo Research, Irvine, CA, USA), which was further used for bacterial genomic DNA isolation. Total bacterial DNA was isolated by Quick-DNA Fecal/Soil Microbe Kits (Zymo Research, Irvine, CA, USA) following the protocol of the manufactures.