|
| Status |
Public on Sep 20, 2012 |
| Title |
fhm F Liver recovery DES 1-4 |
| Sample type |
RNA |
| |
|
| Source name |
liver from female fish exposed to 1 ng DES/L for 96 h, followed by an additional 96 h in control water
|
| Organism |
Pimephales promelas |
| Characteristics |
ctid: 160554
treatment: 1.0 ng DES/L for 4 days then recovery for another 4 days
tank: 6
Sex: F
FISH wet wt (g): 1.13
gonad wt (g): 0.0415
time sampled: 1:05
gsi: 3.67256637168142
liver vtg mrna: 1.08E+04
liver esr1 mrna: 47.02
ex vivo e2 (ng/ml/12h): 0.285499994579278
ex vivo t (ng/ml/12h): 0.0469066053844446
plasma e2 (ng/ml): 1.63634169139705
plasma vtg: 15.5
tissue: liver
|
| Treatment protocol |
Fish were exposed to 0, 1, 10, or 100 ng DES/L delivered in a continuous flow (45 ml/min) of sand filtered, UV treated, Lake Superior Water, without the use of carrier solvents for a total of 96 h. Delivery of DES then ceased and fish were held in a continuous flow (45 ml/min) of control water for an additional 96 h (4 d depuration).
|
| Growth protocol |
The fish were held at 25+1ºC under a 16:8 L:D photoperiod and were fed frozen adult brine shrimp twice daily (San Francisco Bay Brand, Newark, CA, USA). Fish used for the experiment were from an on-site culture, and all procedures involving animals conformed to guidelines approved by the local Animal Care and Use Committee.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from liver tissue with Tri Reagent (Sigma, St. Louis, MO) according to the manufacturer’s protocol. RNA quality was verified using an Agilent 2100 bioanalyzer (Agilent, Palo Alto, CA) and concentration was quantified spectrophotometrically using a Nanodrop ND-1000 spectrophotometer (Nanodrop Technologies, Wilmington, DE).
|
| Label |
cy3
|
| Label protocol |
cDNA synthesis, cRNA labeling, amplification and hybridization were performed following the manufacturer's kits and protocols (Quick Amp Labeling Kit; Agilent, Palo Alto, CA)
|
| |
|
| Hybridization protocol |
The Agilent one-color microarray hybridization protocol (One-Color Microarray-Based Gene Expression Analysis, version 5.7, Agilent Technologies, Palo Alto, CA) was used for microarray hybridizations following the manufacturer’s protocol and recommendations.
|
| Scan protocol |
An Axon GenePix® 4000B Microarray Scanner (Molecular Devices Inc.) was used to scan microarray images at 5 μm resolution
|
| Data processing |
Microarray image processing and data pre-processing were performed Agilent's Feature Extraction software v 9.5 following the manufacturer’s protocol and recommendations.
|
| |
|
| Submission date |
Mar 13, 2012 |
| Last update date |
Sep 20, 2012 |
| Contact name |
Daniel L. Villeneuve |
| E-mail(s) |
villeneuve.dan@epa.gov
|
| Organization name |
US EPA
|
| Department |
Mid-Continent Ecology Division
|
| Lab |
National Health and Environmental Effects Research Laboratory
|
| Street address |
6201 Congdon Blvd
|
| City |
Duluth |
| State/province |
MN |
| ZIP/Postal code |
55804 |
| Country |
USA |
| |
|
| Platform ID |
GPL10259 |
| Series (2) |
| GSE36466 |
A Short-Term Study Investigating the Estrogenic Potency of Diethylstilbesterol in the Fathead Minnow (Pimephales promelas): Recovery phase |
| GSE36467 |
A Short-Term Study Investigating the Estrogenic Potency of Diethylstilbesterol in the Fathead Minnow |
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