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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Regulation of tetrahydrobiopterin biosynthesis in cultured dopamine neurons by depolarization and cAMP.

Primary cultures containing embryonic rat brain mesencephalic or hypothalamic dopamine neurons were used to examine the effects of membrane depolarization and elevations of cAMP levels on tetrahydrobiopterin cofactor content. Initial studies showed that 24-h incubations with 8-bromo-cAMP or isobutyl methylxanthine increased cofactor levels in either culture system, whereas the stimulatory effects of forskolin or depolarization of membrane potential were only observed in cultures of hypothalamus. 8-Bromo-cAMP was found to increase cofactor content in a concentration-dependent manner, with increases observed up to 5 mM. The time course of the effect of 8-bromo-cAMP was biphasic. Over the short term, an increase of 50% in cofactor content at 2 and 5 h was detected. Over the long term, by 24-48 h, cofactor levels increased by between 100% and 300%. Studies of cofactor turnover indicated that the long-term increase was due to stimulation of tetrahydrobiopterin biosynthesis with no alteration in degradation rate. Inhibitors of gene transcription and translation prevented the long- but not short-term increase in cofactor content. Levels of GTP cyclohydrolase I mRNA were increased 7-10-fold following 5 h of incubation with 8-bromo-cAMP. Tetrahydrobiopterin biosynthesis within cultured dopamine neurons of the hypothalamus and mesencephalon thus appears to be regulated by a cAMP-dependent mechanism involving enhanced gene expression of enzyme(s) involved in cofactor biosynthesis.[1]


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