Western blotting for detection of glucocorticoid receptors in the brain and pituitary gland from adrenal intact pigs

摘要: The objectives of the experiments presented here were to develop a method for measuring total glucocorticoid receptors (GRs), in adrenal intact pigs, and describe distribution this receptor pituitary gland selected brain regions. Six male 14 days age, euthanized gland, hypothalamus, prefrontal cortex, hippocampus prepared western blotting using traditional soluble cytosolic preparation or whole cell extract preparation. Samples electrophoresed immunoblotted anti-human GR antibodies #57 #51. antibody labeled doublet at 95 87 kDa whereas #51 single band kDa. Affinity labeling with [(3)H] dexamethasone mesylate revealed Using concentrations highest followed by hypothalamus (P<0. 0001). amount cortex was equivalent but lesser (P<0.0001) than that found hypothalamus. With tissue preparation, (P<0.05) lower levels detected Taken together, these results indicated 87-kDa functional regional GRs is specific type used. In addition, we have described procedure (from extracts) neural nonneural tissues from pigs.