Translatome_Group_B_Rep_1
Mus musculus
Protocols: We got the mice from Shanghai SLAC Laboratory Animal (C57BL/J6, Shanghai, China) and separated the mice into three groups randomly. The mice stayed in a temperature and humidity-controlled environment with free access to standard rodent food and water. The mice received sevoflurane anesthesia using the protocol from Lu et al 2017 (PMID: 28968276). Briefly, one group of mice (defined as Group A) received 2.5% sevoflurane with 60% O2 at the flow rate of 2 L/min for 3 min and 1 L/min for 2 h on postnatal day 6, 7, 8, 9 in an anesthetizing chamber (Datex Ohmeda, Madison, USA). Another group of mice (defined as Group B) received 2.5% sevoflurane with 60% O2 at the flow rate of 2 L/min for 3 min and 1 L/min for 2 h on postnatal day 6, 7 in the same anesthetizing chamber. The control group received 60% O2 at the same flow rate and in the same anesthetizing chamber. Polysome fraction was isolated with the method from Lupinacci et al 2019 (PMID: 31052505) with small changes. Briefly, we lysed the hippocampus tissues in a Polytron with 400 uL lysis buffer (using 100 μg/mL cycloheximide), incubated the lysates on ice for 10 min and conducted centrifugation at 10000x g for 10 min at 4°C. The supernatants were collected and loaded into 10–50% sucrose gradients. Gradients were placed in a Beckman SW41Ti rotor and centrifuged at 39,000 rpm at 4 °C for 2.5 h, after which the polysome fractions were collected. Then, polysome-associated RNA was extracted with TRIZOL according to the manufacturer's protocol (Invitrogen, Waltham, USA). Total RNA was extracted directly from the hippocampus tissues following the same TRIZOL protocol. The extracted RNA was treated with DNase to remove genomic DNA contamination. Isolation of mRNA was based on NEBNext PolyA mRNA Magnetic Isolation Module (New England Biolabs, Ipswich, USA) and the mRNA was further used with NEB Next Ultra Directional RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, USA) for sequencing library preparation.
ENA-FIRST-PUBLIC
2023-03-09
ENA-LAST-UPDATE
2023-03-09
External Id
SAMEA8218461
INSDC center alias
Department of anesthesiology, The first affiliated hospital of Zhengzhou university
INSDC center name
Department of anesthesiology, The first affiliated hospital of Zhengzhou university
INSDC first public
2023-03-09T00:16:28Z
INSDC last update
2023-03-09T00:16:28Z
INSDC status
public
Submitter Id
E-MTAB-10202:Translatome_Group_B_Rep_1
age
36
broker name
ArrayExpress
common name
house mouse
compound
oxygen; 60 percent, sevoflurane; 0.25 percent
developmental stage
neonate
disease
normal
dose
1
genotype
wild type genotype
organism part
hippocampus
sample name
E-MTAB-10202:Translatome_Group_B_Rep_1
scientific_name
Mus musculus
strain
C57BL/J6
time
2
treatment
treated on days 6 to 7