Immobilization Stress Elevates GTP Cyclohydrolase I mRNA Levels in Rat Adrenals Predominantly by Hormonally Mediated Mechanisms

GTP cyclohydrolase I (GTPCH) is the rate-limiting enzyme in the biosynthesis of tetrahydrobiopterin, the cofactor for catecholamine, indolamine and nitric oxide biosynthesis. In this study we examined the effect of immobilization stress on GTPCH mRNA levels and the mechanism(s) of stress-induced changes in adrenomedullary GTPCH mRNA levels. We used reverse-polymerase chain reaction to isolate and clone a cDNA corresponding to nucleotides 269 to 570 of rat GTPCH. Northern blot analysis with a cRNA probe revealed two species of GTPCH mRNA (about 3.6 and 1.2 kb) in rat adrenal medulla and cortex, and in PC12 cells. The levels of both forms of GTPCH mRNA were significantly increased 3-5 fold in adrenal medulla by a single 2 hour immobilization and by repeated immobilizations (2 hours a day for 2 days). Hypophysectomy had little effect on their basal levels but prevented the stress elicited rise in both GTPCH mRNAs. In contrast, unilateral transection of the splanchnic nerve did not affect induction of the 3.6 kb GTPCH mRNA by stress. Combined denervation with hypophysectomy completely blocked the induction of both GTPCH mRNA species by immobilization stress. Thus, stress elicits elevation of both forms of GTPCH mRNA by a mechanism requiring an intact pituitary-adrenocortical axis.