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Sample GSM85194 Query DataSets for GSM85194
Status Public on Mar 20, 2006
Title 3 dpa con b
Sample type RNA
 
Source name Adult Male Zebrafish Caudal Fin 3 Day Post Amputation, Vehicle Exposed Replicate B
Organism Danio rerio
Characteristics strain: AB Zebrafish
age: Adult
gender: Male
time point: 3 day post amputation of caudal fin
exposure: vehicle
Treatment protocol Adult male zebrafish (Danio rerio) (AB strain, Eugene, OR) were anesthetized with tricaine methanesulfonate (MS-222) and their caudal fins were amputated at the bifurcation of the fin rays. Fish were then injected with phosphatidylcholine liposomes (vehicle control) alone or liposomes incorporated with TCDD (>99% pure, Chemsyn, Lenexa, KS) resulting in a final dose of 50 ng TCDD/g fish. At the indicated dpa, fish were euthanized with an over dose of MS-222. Regenerated fin tissue was then isolated by amputation within two ray segments anterior to the original amputation plane. Amputated tissue was then stored in RNAlater (Qiagen, Valencia, CA) and frozen at -80oC until RNA was isolated from the tissue. Tissues from 10 fish were combined to comprise a single experimental group (replicate). Three groups of tissue were obtained from vehicle control or TCDD exposed fish at 1, 3, and 5 dpa.
Growth protocol Fish were matained at 27oC until under standard conditions.
Extracted molecule total RNA
Extraction protocol Amputated tissue was stored in RNAlater (Qiagen, Valencia, CA) and frozen at -80oC. RNAlater was removed from the samples and total RNA was purified with TRI reagent (Molecular Research Laboratories, Cincinnati, OH) according to the manufacturer’s instructions. Total RNA was DNase treated with RQ1 DNase (Promega, Madison, WI) according to the manufacture’s protocol and RNA quantity and quality was determined by UV absorbance. Ribosomal RNA abundance and degree of degradation was determined in electropherogram patterns using the 2100 Bioanalyzer and RNA 6000 Nano chips (Agilent Technologies, Palo Alto, CA).
Label Labeling and probe processing was conducted under manufactures recommendations.
Label protocol 2.5 µg of total RNA was used to generate biotinylated complementary RNA (cRNA) for each treatment group using the One-Cycle Target Labeling kit (Affymetrix, Santa Clara, CA). RNA was reverse transcribed using a T7-(dT)24 primer and Superscript II reverse transcriptase (Invitrogen, Carlsbad, CA).
 
Hybridization protocol Hybridization was conducted as stipulated by Affymetrix GeneChip Expression Analysis Technical Manual (701021 Rev. 5).
Scan protocol Affymetrix fluidics station 400 was used to wash the arrays. Arrays were scanned with an Affymetrix scanner 3000.
Description Adult male zebrafish were i.p. injected with 50 ng/g TCDD or vehicle and caudal fins were amputated. Regenerating fin tissue was collected at 1, 3 and 5 days post amputation for mRNA abundance analysis.
Data processing Data was GC-RMA preprocessed and each gene was normalized to the median using Gene Spring 7.1 software.
 
Submission date Nov 23, 2005
Last update date Jun 13, 2011
Contact name Robert L Tanguay
E-mail(s) robert.tanguay@oregonstate.edu
Phone 541-737-6514
Organization name Oregon State University
Department EMT
Street address 1007 ALS
City Corvallis
State/province OR
ZIP/Postal code 97331
Country USA
 
Platform ID GPL1319
Series (1)
GSE3667 1, 3, or 5 Day Post Amputation Vehicle or TCDD Exposed
Relations
Reanalyzed by GSM269738

Data table header descriptions
ID_REF
VALUE signal intensity after GC-RMA preprocessing and each gene is normalized to the median

Data table
ID_REF VALUE
AFFX-BioB-5_at 97.09397
AFFX-BioB-M_at 181.94836
AFFX-BioB-3_at 77.551346
AFFX-BioC-5_at 187.15363
AFFX-BioC-3_at 177.72617
AFFX-BioDn-5_at 1073.3608
AFFX-BioDn-3_at 1584.5161
AFFX-CreX-5_at 5717.888
AFFX-CreX-3_at 9845.829
AFFX-DapX-5_at 80.95248
AFFX-DapX-M_at 237.6066
AFFX-DapX-3_at 374.36597
AFFX-LysX-5_at 18.194897
AFFX-LysX-M_at 19.620106
AFFX-LysX-3_at 60.58026
AFFX-PheX-5_at 23.682531
AFFX-PheX-M_at 28.95712
AFFX-PheX-3_at 70.11621
AFFX-ThrX-5_at 28.319597
AFFX-ThrX-M_at 22.017843

Total number of rows: 15617

Table truncated, full table size 402 Kbytes.




Supplementary file Size Download File type/resource
GSM85194.CEL.gz 2.2 Mb (ftp)(http) CEL
Raw data provided as supplementary file
Processed data included within Sample table

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