characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-06-06T00:15:06Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-06-06T00:15:06Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA110170652
characteristics.characteristic: External Id
characteristics.text: University of California Irvine
characteristics.characteristic: INSDC center name
characteristics.text: 2023-06-06T00:15:06Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-06-06T00:15:06Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS12269466
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-11848:WT 6-OHDA 2
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Experiments (1) were performed in WT and iMSN-D2RKO littermates unilaterally subjected either to sham or 6-OHDA lesions in the right DLS. Three weeks after surgeries, mice received a daily injection of L-DOPA (15 mg/kg; s.c.) plus benserazide (12 mg/kg; s.c.) for 11 consecutive days.The eleventh day, mice were sacrificed after 60 minutes from L-DOPA or Saline treatment. Mice of both genotypes were sacrificed and brains were flash frozen. Punches of the DLS were obtained on cryostat sections Experiment was performed in WT and iMSN-D2RKO littermates unilaterally subjected either to sham or 6-OHDA lesions in the right DLS. Three treatments groups: 1)Sham mice = mice received surgery without 6-OHDA compound + 11 days of Saline 2)6-OHDA mice= mice received surgery with 6-OHDA compound + 11 days of Saline 3)L-DOPA mice = mice received surgery with 6=OHDA compund + L-DOPA 15 mg/kg; (s.c.) plus benserazide (12 mg/kg; s.c.) Brain Punches were rapidly homogenized in Trizol (Thermo Fisher) using a 26 1⁄2 gauge needle attached to a 1 mL syringe. RNA isolation was completed following the Trizol (Thermo Fisher) manufacturer’s protocol. RNA was resuspended in H2O. Library preparation and sequencing were performed at the University of California, Irvine Genomic High-Throughput Facility. Total RNA was monitored for quality control using the Nanodrop absorbance ratios for 260/280nm and 260/230nm and the RIN (RNA Integrity Number) was evaluated with Agilent Bioanalyzer Nano RNA chip. Library construction was performed according to the Illumina TruSeq Stranded mRNA Sample Preparation Guide. The input quantity for total RNA was 150 ng and mRNA was enriched using oligo dT magnetic beads. The enriched mRNA was chemically fragmented for three minutes. The first strand synthesis used random primers and reverse transcriptase to make cDNA. After second-strand synthesis, the double-stranded cDNA was cleaned using AMPure XP beads and the cDNA was end-repaired and then the 3-ends were adenylated. Illumina barcoded adapters were ligated on the ends and the adaptor-ligated fragments were enriched by nine cycles of PCR. The resulting libraries were validated by qPCR and sized by Agilent Bioanalyzer DNA high sensitivity chip. The concentrations for the libraries were normalized and then multiplexed together.
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: Parkinson disease
characteristics.characteristic: disease
characteristics.text: D2Rflox/flox
characteristics.characteristic: genotype
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: brain
characteristics.characteristic: organism part
characteristics.text: wild type phenotype
characteristics.characteristic: phenotype
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: male
characteristics.characteristic: sex
characteristics.text: WT 6-OHDA 2
characteristics.characteristic: title