|
| Status |
Public on Sep 14, 2011 |
| Title |
Embryonic day 13.5 diabetes replicate 1 |
| Sample type |
RNA |
| |
|
| Source name |
Heart tissue
|
| Organism |
Mus musculus |
| Characteristics |
age: E13.5
strain: Swiss Albino
disease status: diabetes
|
| Treatment protocol |
Diabetes mellitus was induced in 8 week-old female mice by an intraperitoneal injection of streptozotocin (STZ, 75mg/kg body weight, Sigma, USA) dissolved in citrate buffer (0.01 M, pH 4.5) for three successive days. Blood glucose level was examined one week after STZ injection using Glucometer Elite (Bayer, USA). Mice with non-fasting blood glucose level exceeding 16 mmol/l were used as experimental diabetic mice. In contrast, control mice maintained the normal blood glucose levels (4– 6 mmol/l) before and during pregnancy. Timed mating was carried out by placing four female mice with one normal male mouse in a cage overnight. The day when a copulation plug was observed was counted as embryonic day 0.5 (E0.5). Time-mated pregnant mice were divided into two groups control and diabetes. At E13.5 and E15.5, pregnant mice were anaesthetized with pentobarbital (150 mg/kg body wt, intraperitoneally) and embryos were collected after Caesarean. For each experiment, hearts were isolated from embryos from the two groups at different time points. The developmental stage, E13.5 corresponds to ~6 weeks of human gestation, hence it was chosen for this study. At this stage all 4 chambers of the heart were clearly distinguishable, and the outflow tract was evident. In addition, the heart phenotype was apparent in embryos from diabetic mice. The embryos with cardiac malformations from diabetic mice were classified as diabetic group and normal embryos from diabetic and non-diabetic mice were used as control groups.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from developing hearts was extracted using RNeasy mini kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
The RNA samples were processed according to the Affymetrix 3’ IVT Express protocol and Origen Lab’s SOP for 3’ IVT Express analysis. Briefly, 100 ng of total RNA was reverse transcribed to produce cDNA, which was subsequently used as a template to create biotin- labeled aRNA (amplified RNA).
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| |
|
| Hybridization protocol |
The labeled cRNA was then cleaned up, fragmented, and hybridized to Mouse Genome 430 2.0 Array (Affymetrix, CA, USA).
|
| Scan protocol |
Arrays were using an Affymetrix 3000 7G scanner.
|
| Description |
Gene expression data from heart tissue isolated from embryonic day 13.5 diabetes exposed mouse
|
| Data processing |
CEL files were exported to Affymetrix Expression Console Version 1.1 and RMA normalization (using EC1.1) was carried out.
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| |
|
| Submission date |
Sep 13, 2011 |
| Last update date |
Sep 14, 2011 |
| Contact name |
Srinivasan Dinesh Kumar |
| E-mail(s) |
[email protected]
|
| Phone |
+65-65923055
|
| Fax |
+65-67908140
|
| URL |
http://scholar.google.com/citations?hl=en&user=FgC87RUAAAAJ
|
| Organization name |
Nanyang Technological University
|
| Department |
Lee Kong Chian School of Medicine
|
| Street address |
50 Nanyang Drive, Research Techno Plaza
|
| City |
Singapore |
| ZIP/Postal code |
637553 |
| Country |
Singapore |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE32078 |
Differential gene expression profiles during embryonic heart development in diabetic mice pregnancy |
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