Healthy male Wistar Kyoto (WKY) rats, 12 weeks old, were purchased from Charles River Laboratories Inc., Raleigh, NC. Animals were double housed in polycarbonate cages with beta chips bedding and acclimatized for at least one week in an AAALAC-approved animal facility (21 ± 1OC, 50 ± 5% relative humidity, 12:12 h light/dark cycle) prior to the experimental period. All animals received standard Purina rat chow (Brentwood, MO) and water ad libitum, except during exposures.Rats were randomly assigned to four groups (n = 15 per group) according to body weight, and exposed by nose-only inhalation (Ledbetter et al., 1998) to either filtered air or aerosolized zinc sulfate (10, 30 or 100 ug Zn/m3) for 5 h/day, 3 days/week for 16 weeks. The aerosol was generated by aerosolizing solutions of zinc sulfate heptahydrate (Sigma Aldrich Chemicals), mixed at different starting concentrations with a TSI Model 3076 atomizer operating at 30 PSIG. The output was mixed with dry air and directed to the exposure chamber (air flow of 98-105 liters/minute). Chamber concentration data was determined by gravimetric analysis, with Inductively Coupled Plasma-Optical Emission Spectroscopy (ICP-OES) to determine zinc concentration.
Extracted molecule
total RNA
Extraction protocol
Target RNA was isolated using the RNeasy Mini Kit (Qiagen, Valencia, CA) and further purified using the RNeasy MinElute Cleanup Kit (Qiagen, Valencia, CA).
Label
biotin
Label protocol
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 6 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
Hybridization protocol
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Drosophila Genome Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
Scan protocol
GeneChips were scanned using Affymetrix Scanner 3000G
Description
zincsulfate-30ug-heart-45
Data processing
The data were analyzed with Rosetta Resolver 6.0 using Affymetrix default profile builder analysis settings.
