E60T-rep2
Rattus norvegicus
Protocols: Primary hypothalamic neurons were prepared from embryonic day 18 (E18) Sprague-Dawley rats. Briefly, hypothalamus was rapidly dissected under sterile conditions, minced into small fragments, and digested using Pierce Primary Neuron Isolation Kit. The digestion was terminated by adding an equal volume of B-27 Neurobasel Plus Medium. Cells were gently triturated using a fire-polished Pasteur pipette, passed through a 70-µm cell strainer, and plated onto 100 µg/mL poly-D-lysine-coated culture dishes at a density of 1 × 105 cells/cm². After 4 h, the medium was replaced. Cultures were maintained at 37 °C in a humidified incubator with 5% CO₂, and half of the medium was replaced every 3 days. To keep the Kiss1 gene activity, 0.1 nM E2 was always added in medium. 1 nM (final concentration, similarly hereinafter) E2 for 24 h or 60 h to model the short and long term estrogen exposure, then for additional 48 h culturing after E2 withdrawal. Total RNA of 5 x 107 cells were extracted using TRIZOL (Thermo Fisher Scientific). Nanodrop 2000 (Thermo Fisher Scientific) and Agilent Bioanalyzer 2100 (Agilent) were used to evaluate RNA concentration and quality. 5 μg of RNA from different experimental groups was prepared cDNA library using the NEBNext Ultra Directional RNA Library Prep Kit for Illumina (NEB) following the manufacturer's instructions.
ENA first public
2026-02-02
INSDC center name
Suzhou Ninth People's Hospital
INSDC status
public
Submitter Id
E-MTAB-16415:E60T-rep2
broker name
ArrayExpress
cell line
primary cell line
cell type
neuron
collection date
not collected
common name
Norway rat
developmental stage
embryo stage
disease
Null syndrome
genotype
wild type genotype
geographic location (country and/or sea)
not collected
isolate
not applicable
organism part
hypothalamus
sample name
E-MTAB-16415:E60T-rep2
scientific_name
Rattus norvegicus
stimulus
treatment