|
| Status |
Public on Oct 15, 2007 |
| Title |
Frontal_Cerebrocortex_Permethrin_10mg/kg_6hr_rep6 |
| Sample type |
RNA |
| |
|
| Source name |
Frontal Cerebrocortex, permethrin (10mg/kg), 6 hours
|
| Organism |
Rattus norvegicus |
| Characteristics |
Strain: Long-Evans
Gender: Male
Age: Adult (PND62-PND64)
Tissue: Cerebrocortex
|
| Biomaterial provider |
Charles River, Inc.
|
| Treatment protocol |
Test Compound: Permethrin
Test Compound CAS#: 52645-53-1
Isomer Composition: 40%-cis, 60%-trans, 1:1 ratio of 1R, 1S
Purity: 92.0 %
Vehicle: Corn Oil
Vehicle CAS #: 8001-30-07
Administration Route: Oral Gavage
Test Compound Concentration: 10 mg/mL
Delivery Volume: 1 mL/kg body weight
Expsoure Duration: Single, Acute
Sampling Time: 6 Hours post-exposure
|
| Growth protocol |
Test subjects housed 2 per cage in polycarbonate hanging cages (45cm X 24 cm X 20 cm) with heat sterilized pine shaving for bedding. Subjects maintained on a 12h:12h photoperiod (06:00-18:00) and allowed a 5-7 day period of acclimation to holding suite prior to treatment. Holding suite maintained at 22.0 +/- 2.0 C with a relative humidity of 55 +/- 20%. Purina 5001 Rat Chow and tap water were provided ad libitum.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from cortical tissue using TRI Reagent (Molecular Research Center, Inc., Cincinnati, OH) per manufacturer's instructions. Following TRI Reagent extraction, total RNA suspended in DEPC-H20 was treated with 16 units of RNase-free DNase I (Ambion, Inc., Austin, TX) and incubated at 37 C for 1 hour. DNase I was inactivated with 1 volume of 5:1 acid-phenol:chloroform, pH=4.7. Samples were vortexed for 60s, stored at 25 C for 15 min and centrifuged for 15 min. Aqueous phase was removed, supplemented with 1 volume of chloroform, vortexed for 60 s, stored at 25 C for 15 min and centrifuged for 15 min. Aqueous phase was removed and total RNA precipitated overnight with 0.1 volumes of 3 M sodium acetate (pH = 5.2) and 2.5 volumes of 100 % ethanol following by centrifugation for 15 min. Precipitated pellets samples were washed with 75% ethanol and resuspended in DEPC H2O.
|
| Label |
biotin
|
| Label protocol |
First and second strand cDNA synthesis, RNase H digestion and (ds)cDNA isolation were performed according to Affymetrix protocol (Affymetrix pub #701021). Biotin-labeled cRNA was synthesized with BioArray HighYeild RNA transcript labeling kit (Enzo Life Sciences, Farmingdale, NY) using manufacturer's protocol. Clean up was performed with Qiagen RNeasy spin columns (Spoorstraat, Netherlands) according to manufacturer's protocol. Biotin-labeled cRNA was fragmented using Affymetrix 5X fragmentation buffer.
|
| |
|
| Hybridization protocol |
Hybridization of Affymetrix Rat 230 2.0 GeneChips was performed according to manufacturer's instructions (Affymetrix pub #701021).
|
| Scan protocol |
GeneChips were scanned using an Affymetrix GeneChip 3000 Scanner with the Affymetrix GCOS v1.2 software package.
|
| Description |
This sample is part of a data set examining dose-related effects of pyrethroid insectcides on gene expression in the mammalian nervous system.
|
| Data processing |
Signal intensities calculated using RMA algorithm
|
| |
|
| Submission date |
May 29, 2007 |
| Last update date |
Sep 13, 2007 |
| Contact name |
Joshua A Harrill |
| E-mail(s) |
[email protected]
|
| Phone |
919-541-4606
|
| Organization name |
University of North Carolina
|
| Department |
Curriculum in Toxicology
|
| Lab |
NBTB/NTD/NHEERL/ORD/USEPA
|
| Street address |
109 TW Alexander Drive
|
| City |
Research Triangle Park |
| State/province |
NC |
| ZIP/Postal code |
27711 |
| Country |
USA |
| |
|
| Platform ID |
GPL1355 |
| Series (1) |
| GSE7955 |
Transcriptional response of rat cerebrocortical tissue following acute exposure to permethrin or deltamethrin. |
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