Dataset features


Application: Gene expression microarray analysis
Number of samples: 12
Release date: Jun 30 2009
Last update date: Mar 20 2012
Access: Public
Diseases: Phenylketonurias
Chemicals: Metyrapone
Dataset link Pituitaries of basal and stressed Crhr1 wild type and knockout mice

Experimental Protocol

To activate the CRHR1-dependent signaling in the pituitary of CRHR1-wt and -ko mice animals were stressed using 30 min restraint stress in 50 ml Falcon tubes. 12 h before stress mice were injected intraperitonally (i.p.) with 150 mg/kg bodyweight (b.w.) metyrapone. Metyrapone is an inhibitor of the steroid beta-hydroxylase, an enzyme catalyzing the biosynthesis of corticosterone. By this way, the negative feedback of corticosterone on the HPA-axis was blocked. A second metyrapone injection (100 mg/kg b.w.) was applied directly before stress. 1 and 3h, respectively, after stress, mice were anesthetized, decapitated and the pituitaries prepared and flash-frozen. mRNA was isolated according to the TRIzol protocol (Invitrogen). Pooled amplified RNA samples were then hybridised on Illumina mouse BeadChips (N=2 per group) and detected in the Illumina BeadArray Reader (Illumina, Inc., San Diego, CA). The experiment was performed in technical duplicates.