Figure 1. Mouse dermal fibroblasts express Crh and its receptors.

(a) Total RNA isolated from whole brain from Crh+/+ mice (lane 1), NMF isolated from Crh+/+ mice (lane 2) and NMF isolated from Crh−/− mice (lane 3) was subjected to RT-PCR for evaluation of Crh expression. Lane 4 represents a sample without RT-enzyme and lane 5 represents a sample without template. (b) Crf1 mRNA expression in NMF isolated from Crh+/+ mice (lane 2). Lane 1 represents Crf1 mRNA expression isolated from whole brain, while lane 3 represents a sample without RT-enzyme and lane 4 a sample without template. (c) Crf2 mRNA expression in NMF isolated from Crh+/+ mice (lane 2). Lane 1 represents Crf1 mRNA expression isolated from heart, while lane 3 represents a sample without RT-enzyme and lane 4 a sample without template. (d) Effects of antalarmin and astressin-2B on specific [¹²⁵I] Tyr0-sauvagine binding to CRF₁ and CRF₂. Membrane homogenates from Crh+/+ and Crh−/− fibroblasts were assayed for specific binding with [¹²⁵I] Tyr0-sauvagine, as described in Materials and Methods. The bars represent the % decrease of specific binding. The mean ± SEM values are from 3 independent experiments, each performed with duplicate determinations. (e) Effect of CRH on cAMP accumulation in Crh+/+ and Crh−/− fibroblasts. Stimulation of cAMP accumulation by CRH was performed as described in Materials and Methods in intact cells. The mean ± SEM values are from 3 independent experiments, each performed with duplicate determinations. * represents statistical difference (P<0.05) between genotypes exposed to the same treatment and # represents statistical difference (P<0.05) between different treatments in the same genotype.