|Application:||Gene expression microarray analysis|
|Number of samples:||12|
|Release date:||Dec 31 2006|
|Last update date:||Dec 6 2012|
|Diseases:||Cardiomyopathies, Thrombosis, Chemical and Drug Induced Liver Injury|
|Dataset link||Bis(2-chloroethoxy)methane-induced heart toxicity and recovery in male fisher rats|
Chemical and Animal exposures: Bis(2-chloroethoxy)methane (CAS No. 11-91-1; lot B004160277) (Karl Industries, Aurora, OH) (Fig, 1) was found to be 98.5% pure (Dunnick et al. 2004a; Dunnick et al. 2004b) . Solutions of BCEM were prepared in 95% ethanol for dosing by dermal administration daily to male F344 rats, excluding weekends, for two weeks plus two consecutive dosages before the last sacrifice on study-day 16, at which animals had received a total of 12 doses. Fur from the site of application was clipped weekly. Stock solutions prepared at concentrations of 0, 800, and 1200 mg/ml were stored in amber glass bottles at room temperature. The administrations were applied to the skin of the male rats at 0.5 ml/kg body to deliver doses of 0, 400, or 600 mg/kg body weight. All dose formulations were determined to be within ±10% of target concentrations. Approximately 45% of a dermal dose of CEM is adsorbed (NIEHS Contract NO1-ES-75407 2002). Male F344/N rats (Taconic Laboratories, Germantown, NY) were placed on study at 5 weeks of age and housed one per cage in polycarbonate cages in rooms maintained at temperatures between 69 and 75°F with 35 – 65% relative humidity and a 12-hr light/dark cycle. Control and treated groups received irradiated NTP-2000 diet (Zeigler Brothers, Gardners, PA) ad libitum. Histopathology: Animals were anesthetized with CO 2 and heart collected and preserved in 10% formalin. Slides were made from paraffin sections from rats treated with BCEM as previously described (Dunnick et al. 2004a). Pathologic analysis included comparing heart lesions in male rats after 2, 3, 5, and 16 days of treatment. RNA Extraction Methods RNA was extracted from hearts of three control animal at day 2, three control rats at day 5, three rats exposed to 200, 400, or 600 mg/kg BCEM at day 2 and three at day 5. Animals designated for heart RNA extraction were anesthetized with CO2/O2 on study days 2 and 5, exanguinated, and their hearts infused with RNAlater. Total cardiac RNA was isolated from hearts using the QIAGEN Rneasy kit (QIAGEN, Valencia, CA). The RNA was quantified through optical density measurements and agarose gel electrophoresis, and kept frozen at – 70 C. RNA was extracted from control, 200, 400, and 600 mg/kg rats.