|
| Status |
Public on Jan 01, 2011 |
| Title |
Day 5 Mouse inguinal fat over nutrition 3 |
| Sample type |
RNA |
| |
|
| Source name |
Mouse age 5 days inguinal fat overnutrition diet
|
| Organism |
Mus musculus |
| Characteristics |
gender: male
strain: C57BL/6J
tissue: inguinal fat
age: 5 days
diet: Lactation over-nutrition condition (LON)- mother was fed a high fat 58 Kcal % fat diet (Research Diet 12331, New Brunswick, New Jersey, United States) ad libitum.
litter size: reduced to 4 pups
|
| Treatment protocol |
See Samples Section for protocol during pregnancy and lactation. After weaning the offspring from the 3 nutritional conditions were treated the same; from weaning until 8 wk of age mice were fed a low fat chow diet (LabDiets 5053 11 Kcal % fat) ad libitum. At 8 wk of age mice were fed ad libitum a high saturated fat diet D12331 (Research Diets,) for 8 weeks. From weaning until 7 wk of age male mice were group housed (3–5 mice per pen) until 7 wk of age, at which time they were singly housed for the remainder of the experiment.
|
| Growth protocol |
At birth, litter sizes were adjusted to 8 pups per litter for control and lactation under-nutrition and 4 pups per litter for lactation over-nutrition with both male and female pups; however, only male pups (12 pups per sample) were sacrificed to form the pools for each time point.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Tissues were quickly removed and frozen in liquid nitrogen and stored at -80C for subsequent preparation of total RNA. Total RNA was isolated using TRI-Reagent (Molecular Research Center Inc. Cincinnati, Ohio) with modifications to remove DNA using the Qiagen RNAeasy columns and DNaseI Kit (Qiagen, Valencia, CA). RNA was stored at –70C in RNase-free H2O supplemented with the RNase inhibitor Superasin (Ambion, Austin, TX) as per manufacturers directions. Quality and quantity of RNA was determined using a Agilent Bioanalyzer as per manufacturers procedures (Agilent Technologies, Palo Alto, CA).
|
| Label |
Dig-UTP
|
| Label protocol |
1 ug of total RNA was used to transcribe DIG-labeled cRNA using Applied Biosystems Chemiluminescent RT-IVT Kit V2.0.
|
| |
|
| Hybridization protocol |
Microarray hybridization 3 technical replicates of each sample (using ten micrograms of fragmented, DIG-labeled cRNA) and processing were performed according to Applied Biosystems protocols.
|
| Scan protocol |
Chemiluminescence detection, imaging, auto gridding, and image analysis was done according to Applied Biosystems protocols and the 1700 Chemiluminescent Microarray Analyzer Software v. 1.0.3.
|
| Description |
D5-LON-9
|
| Data processing |
Signal intensities across microarrays were normalized using the quantile-quantile method (www.bioconductor.org).
|
| |
|
| Submission date |
Jan 08, 2010 |
| Last update date |
Jan 01, 2011 |
| Contact name |
Robert A. Koza |
| E-mail(s) |
kozara@pbrc.edu
|
| Phone |
225-763-3191
|
| Organization name |
Pennington Biomedical Research Center
|
| Department |
Genomics Core Facility
|
| Street address |
|
| City |
Baton Rouge |
| State/province |
LA |
| ZIP/Postal code |
70808 |
| Country |
USA |
| |
|
| Platform ID |
GPL2995 |
| Series (1) |
| GSE19809 |
Efect of neonatal nutrition on adipose tissue remodeling genes during early development and in adult mice |
|
