|
| Status |
Public on Jun 28, 2023 |
| Title |
P30-HS 5 |
| Sample type |
RNA |
| |
|
| Source name |
rat hippocampal ventral CA3
|
| Organism |
Rattus norvegicus |
| Characteristics |
gender: Male
age (days): 30
group: animal with seizure
|
| Treatment protocol |
Fresh ex-vivo explants of rat hippocampal ventral CA3 were obtained from dissected brain and collected after animal sacrifice following 20 days (P30) from the hyperthermic insult. CA3 samples were immediately kept in 600uL of RNAlater (Qiagen) until RNA extraction.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
CA3 explants (3-4 mm3) were homogenized with TissueRupter (Qiagen) and total RNA was extracted from the homogenates using the RNeasy Lipid Tissue Kit (Qiagen)
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.2 ug RNA using the One-Color Microarray-Based Gene Expression Analysis - Quick Amp Labeling (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (Qiagen). RNA concentration and purity was determined by reading the absorbance at 260 and 280 nm on a spectrophotometer (NanoVue, GE Health Care, USA).
|
| |
|
| Hybridization protocol |
1.5 ug of Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes and hybridized to Rat Gene Expression 4x44K v3 Microarray Kit (G2519F-028282, Agilent Technologies) for 17 hours at 65°C in a rotating Agilent hybridization oven.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent Bundle using one color scan setting for 4x44k array slides.
|
| Description |
Gene expression from rat hippocampal ventral CA3
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 11.5.1.1 (Agilent) and considering spots present none or only one flag (i.e. low intensity, saturation, controls, etc.). The selected transcripts were used for analysis using the R software and considering none NA.
|
| |
|
| Submission date |
Apr 14, 2023 |
| Last update date |
Jun 28, 2023 |
| Contact name |
Carlos Alberto Moreira-Filho |
| E-mail(s) |
[email protected]
|
| Phone |
55-11-3061-8449
|
| Organization name |
Faculdade de Medicina da Universidade de Sao Paulo
|
| Department |
Departments of Pediatrics
|
| Lab |
LIM36
|
| Street address |
Av. Dr. Eneas Carvalho de Aguiar 647
|
| City |
Sao Paulo |
| State/province |
SP |
| ZIP/Postal code |
05403-900 |
| Country |
Brazil |
| |
|
| Platform ID |
GPL14746 |
| Series (2) |
| GSE229754 |
Transcriptomic analysis reveals distinct adaptive molecular mechanism in the hippocampal CA3 from rats susceptible or not-susceptible to hyperthermia-induced seizures |
| GSE229760 |
Transcriptomic analysis reveals distinct adaptive molecular mechanism in the hippocampal CA3 from rats susceptible or not-susceptible to hyperthermia-induced seizures |
|
