title: iMSN-D2RKO L-DOPA 3
organism: Mus musculus
description: 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.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2023-06-06
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-06-06
attributes.tag: External Id
attributes.value: SAMEA110170647
attributes.tag: INSDC center alias
attributes.value: University of California Irvine
attributes.tag: INSDC center name
attributes.value: University of California Irvine
attributes.tag: INSDC first public
attributes.value: 2023-06-06T00:15:06Z
attributes.tag: INSDC last update
attributes.value: 2023-06-06T00:15:06Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-11848:iMSN-D2RKO L-DOPA 3
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: normal
attributes.tag: genotype
attributes.value: D2Rflox/flox/D1-Cre/+
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: brain
attributes.tag: phenotype
attributes.value: iMSN-D2RKO
attributes.tag: sample name
attributes.value: E-MTAB-11848:iMSN-D2RKO L-DOPA 3
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: male