array type: Rat230_2 amount to core: 17 units of amount: ul od260: 3.427 od280: 1.716 dilution: 1:05 rna integrity number: 8.2 bioanalyzer results: good sample preservation: Cells suspended in RNAlater at 4 degrees Centigrade for 24 hours and then frozen at -20 degrees Centigrade until processed for RNA. strain or line: F344/CrlBR ncbi taxonomy id: 10116 individual: 212 developmental state: Adult sacrifice method: Deep pentobarbital anesthesia followed by exsanguination sex: Male units of age: weeks age: 8 tissue: Nasal epithelial cells (level II region) tissue harvest methods: The nose was dissected to isolate the Level II region of both nostrils. The maxillo turbinate and olfactory epithelium were discarded and the lateral meatus and nasoturbinate were retained for cell isolation. The dissected nasal sections were rinsed with cold phosphate buffered saline to remove the blood. After rinsing, a 5 ml buffered mixture of proteases (collagenase, 80 U/ml, Sigma-Aldrich; pronase, 10 mg/ml, Sigma-Aldrich; HEPES, 0.1 M) in Hams F12 medium (Invitrogen, Carlsbad, CA) was added to the nose and incubated at 37°C for 40 minutes to remove the epithelial cell layer. Following incubation, the mixture was vortexed and the resulting cell suspension was collected in a clean centrifuge tube. The cell suspension was centrifuged (200 x g, 10 min, 10°C) and the cell pellet was resuspended in 50 ?L PBS and 200 ?L RNAlater (Ambion, Austin, TX). The epithelial cells acquired consisted primarily of transitional epithelium with some respiratory epithelium. units of harvest time point: weeks time point: 4w agent: formaldehyde dose: 0
Biomaterial provider
Russell Thomas, The Hamner Institutes for Health Sciences, 6 Davis Drive, PO Box 12137, Research Triangle Park, NC, 27709, Russell Thomas, rthomas@thehamner.org, 919-558-1311, 919-558-1300
Treatment protocol
in vivo Treatment: Air Control, Treatment Repeat: 5, Units of Treatment Repeat: days per week, Treatment Duration: 5, Units of Duration: days, Treatment Time: 6, Units of Treatment Time: hours per day, Route of Exposure: Inhalation (whole-body), Dose or Concentration: 0, Units of Dose or Concentration: ppm
Extracted molecule
total RNA
Extraction protocol
Extraction Method: Qiagen Micro RNA kit
Label
biotin
Label protocol
standard Affymetrix protocol
Hybridization protocol
standard Affymetrix procedures
Scan protocol
standard Affymetrix procedures
Description
time point: 4w dose: 0 EB_0_4w_212_ACO2_THI08031_Rat230_2_THI 08031: Eight week old male F344/CrlBR rats were exposed to formaldehyde through whole body inhalation. Whole-body exposures were performed at doses of 0, 0.7, 2, 6, 10, and 15 ppm (6 hours per day, 5 days per week). Inhalation animals were sacrified at 1, 4, and 13 weeks following initiation of exposure. Following sacrifice, tissue from the Level II region of the nose was dissected and digested with a mixture of proteases to remove the epithelial cells. The epithelial cells acquired from this section of the nose consisted primarily of transitional epithelium with some respiratory epithelium. Gene expression microarray analysis was performed on the epithelial cells.
