|
| Status |
Public on Sep 20, 2012 |
| Title |
fhm F liver exposure con-6 |
| Sample type |
RNA |
| |
|
| Source name |
liver from female fish exposed to control water for 96 h
|
| Organism |
Pimephales promelas |
| Characteristics |
ctid: 160454
treatment: control
tank: 3
Sex: F
FISH wet wt (g): 1.17
gonad wt (g): 0.064
time sampled: 1:23
gsi: 5.47008547008547
liver vtg mrna: 7.21E+05
liver esr1 mrna: 128.2
ex vivo e2 (ng/ml/12h): 0.382033726126491
ex vivo t (ng/ml/12h): 0.0266469191567719
plasma e2 (ng/ml): 1.63400354972898
plasma vtg: 2.5
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. Total exposure time was 4 d (96 h).
|
| 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) |
| GSE36465 |
A Short-Term Study Investigating the Estrogenic Potency of Diethylstilbesterol in the Fathead Minnow (Pimephales promelas): Exposure Phase |
| GSE36467 |
A Short-Term Study Investigating the Estrogenic Potency of Diethylstilbesterol in the Fathead Minnow |
|
