|
| Status |
Public on Jun 16, 2014 |
| Title |
OGD 24h_R1 |
| Sample type |
RNA |
| |
|
| Source name |
Oxygen and Glucose Deprivation 24h_R1
|
| Organism |
Rattus norvegicus |
| Characteristics |
strain background: Wistar
develpmetal stage: E18 rat embryos
tissue: Primary hippocampal neurons
exposed to: Oxygen and Glucose deprivation (OGD)
time point: after 24h recovery
|
| Treatment protocol |
For the OGD challenge, hippocampal cultures were incubated in a glucose-free saline buffer (in mM: 10 HEPES, 116 NaCl, 5.4 KCl, 0.8 MgSO4, 1 NaH2PO4, 1.8 CaCl2, 25 NaHCO3, 25 sucrose, pH 7.3) in an anaerobic chamber (Forma Anaerobic System, Thermo Fisher Scientific), at 37C, for the indicated times. Control neurons were placed in a similar saline buffer with 25 mM glucose instead of sucrose and kept in an air/CO2 incubator, at 37C, for the same period of time. After the stimulus periods, the saline buffers were replaced by conditioned medium and the cultures returned to the air/CO2 incubator, where they were kept to recover for the indicated times.
|
| Growth protocol |
The cultures were maintained in Neurobasal medium in a humidified incubator with 5% CO2/95% air, at 37°C, for 14-15 days.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from three rat hippocampal neuronal cultures submitted to control or OGD conditions were collected after 7h and 24h of recovery
|
| Label |
Cy3
|
| Label protocol |
Equal amounts of RNA extract (200 ng) from each replicate were amplified and Cy-3-labeled using the Low Input Quick Amp Labeling kit (Agilent Technologies)
|
| |
|
| Hybridization protocol |
Hybridizations were carried out following Agilent Technologies instructions for One-Color Microarray-Based Gene Expression Analysis (Agilent Technologies, Santa Clara, CA, USA), using whole-genome Rat GE 4x44K v3 Microarrays.
|
| Scan protocol |
Images were obtained using Agilent G2565AA microarray scanner and fluorescence quantization was performed using Agilent Feature Extraction 10.5.1.1 software and GE1_105_Dec08 protocol.
|
| Description |
Gene expression of OGD-stimulated neurons after 24h
|
| Data processing |
The signal intensity was aligned and normalized between microarrays by centering the median of the signal distribution using BRB-ArrayTools v3.8.1.
|
| |
|
| Submission date |
Jan 13, 2014 |
| Last update date |
Jun 16, 2014 |
| Contact name |
Joana Filipa Fernandes |
| E-mail(s) |
[email protected]
|
| Organization name |
Center for Neuroscience and Cell Biology
|
| Department |
Life Sciences
|
| Lab |
Neurobiology and Disease
|
| Street address |
Rua Larga, Faculdade de Medicina 2º andar, lab 206A Coimbra
|
| City |
Coimbra |
| State/province |
-- Please Select -- |
| ZIP/Postal code |
3004-504 |
| Country |
Portugal |
| |
|
| Platform ID |
GPL14746 |
| Series (1) |
| GSE54037 |
In vitro ischemia triggers a transcriptional response to down-regulate synaptic proteins in hippocampal neurons. |
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