Lines A and C are derived from F1 crosses of H/W × L-E rats (Tuomisto et al., 1999). F344 rats are moderately resistant to TCDD but their LD50 values vary depending on the supplier (from 164 to 340 μg TCDD/kg body weight) (Walden and Schiller, 1985). Wis rats, on the other hand, exhibit
a mixed population of AHR genotypes, consisting of either AHRwt/wt, AHRwt/hw, or AHRhw/hw. Wis rats’ sensitivities to TCDD vary Y 27632 according to the genotype that they carry (Kawakami et al., 2009). All the Wis rats employed in the present study were of the homozygous wildtype AHR genotype and are thus more sensitive than H/W rats (see Methods). Our goals here are two-fold. First, we survey for the first time the inter-strain heterogeneity of rat transcriptomic responses to TCDD within a single consistent experiment. Second, we exploit the genetic diversity amongst these rat strains to identify genes that show Type-I and Type-II responses to TCDD. Type-I genes might regulate common dioxin-induced selleck chemicals llc toxicities in both sensitive and resistant rats; Type-II genes are candidates to explain dioxin toxicities unique to sensitive rats and not observed in resistant rats. We hypothesize that the genetic “filter” imposed by inter-strain variability will facilitate identification of candidate genes for AHR-regulated toxicities. Male rats of four strains and two lines were examined: Long-Evans
(L-E), Han/Wistar (Kuopio) (H/W), Fischer 344 (F344), Wistar (Wis), Line-A (LnA) and Line-C (LnC). Animals were either treated with 100 μg/kg TCDD or corn-oil
vehicle (4 mL/kg by gavage) at the age of 11–15 weeks. The treatment dose chosen is lethal to all animals in dioxin-sensitive strains but not to any animals in dioxin-resistant strains ( Fig. 1) ( Pohjanvirta and Tuomisto, 1994, Tuomisto et al., 1999 and Walden and Schiller, 1985). We confirmed that all Wistar animals possessed wild-type AHR by PCR analysis of liver cDNA as previously described ( Pohjanvirta, 2009). The rats were housed singly in stainless steel wire-mesh cages and given access to R36 feed (Ewos, Södertälje, Sweden) and water. Animals were fed during the early light hours daily. Artificial illumination was provided in the rooms with light and dark cycles every 12 h with lights on daily at 07:00. The room temperature check details was maintained at 21.5 ± 1 °C and humidity at 55 ± 10%. In total, 208 animals (56 for microarray only and the remaining 152 for PCR validation) were used. Animals in the microarray experiments were euthanized 19 h after treatment with TCDD or corn oil vehicle. Animals in the time-course experiments were given either 100 μg/kg TCDD or corn-oil vehicle and their liver excised at different time intervals (from 0 to 384 h) and animals in the dose–response experiments were treated with different doses of TCDD (from from 0 to 3000 μg/kg) or corn-oil vehicle and their livers removed at 19 h post-treatment.