|
| Status |
Public on Sep 08, 2014 |
| Title |
leftventricle_I/R-surgery_5d_WT_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
left ventricular myocardium ischemic area, WT, I/R, 5d
|
| Organism |
Mus musculus |
| Characteristics |
strain/background: mixed (129/Sv and C57BL/6)
genotype/variation: wild-type (noCre)
treatment: myocardial ischemia/reperfusion surgery
time point (post-surgery): 5d
tissue: left ventricular myocardium ischemic area
|
| Treatment protocol |
Mice were anesthetized and subjected to myocardial ischemia/reperfusion injury. Briefly, after initiation of general anesthesia with Propofol 1% (0.002 ml/g bodyweight, retroorbital injection) and isoflurane anesthetic gas, the mice were intubated, ventilated with a rodent ventilator (MiniVent Type 845, Harvard Apparatus, Holliston, MA) and a left thoracotomy was performed. Anesthesia was maintained with isoflurane 2.0 %. The left anterior descending (LAD) coronary artery was ligated with an 8-0 suture, tied on top of a plastic tube, 0.9 mm in diameter, approximately 1 mm below the edge of the left atrial appendage. Successful ligation of the artery was confirmed by blanching of the left ventricular (LV) myocardium. After 60 minutes of ischemia, reperfusion was established by pulling out the occluding tube. 24 hours or 5 days post-surgery, mice were euthanized and hearts were harvested to obtain RNA.
|
| Growth protocol |
Mixed mouse background strain: 129/Sv and C57BL/6 with CaMKII delta/gamma double KO.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from ventricular tissue or from cultured cardiomyocytes using TRIzol (Invitrogen, Germany). Total RNA was digested with DNase, and cDNA synthesis from 500 ng of RNA was carried out using a SuperScript first-strand synthesis system for RT-PCR (Invitrogen).
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol.
|
| |
|
| Hybridization protocol |
Hybridization (16h x 45°C) was processed according to the standard Affymetrix protocol.
|
| Scan protocol |
Affymetrix GeneArray Scanner 3000.
|
| Description |
noCre_5d_7
I/R = myocardial ischemia/reperfusion surgery
|
| Data processing |
The data were analyzed with a commercial software called JMP Genomics, version 5, from SAS. Gene expression profiling was performed using arrays of mogene20st -type from Affymetrix. A Custom CDF Version 18 with Entrez-based gene definitions was used to annotate the arrays. The Raw fluorescence intensity values were normalized applying quantile normalization, RMA background correction and Medianpolish Probeset Summary.
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| |
|
| Submission date |
Jun 13, 2014 |
| Last update date |
Sep 08, 2014 |
| Contact name |
Carsten Sticht |
| Organization name |
University Heidelberg
|
| Department |
ZMF
|
| Street address |
Theodor-Kutzer-Ufer
|
| City |
Mannheim |
| ZIP/Postal code |
68169 |
| Country |
Germany |
| |
|
| Platform ID |
GPL18802 |
| Series (1) |
| GSE58486 |
CaM Kinase II mediates maladaptive post-infarct remodeling but not acute myocardial ischemia/reperfusion injury |
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