Simultaneous measurement of tyrosine and tryptophan hydroxylase activities in brain in Vivo using an inhibitor of the aromatic amino acid decarboxylase

Summary

DOPA and 5-HTP accumulated in vivo in rat brain after decarboxylase inhibition with NSD 1015 (3-hydroxybenzylhydrazine). This accumulation was linear for the first 30 min and occurred in several brain regions over a wide range of NSD 1015 doses. After a peripheral decarboxylase inhibitor much less, if any, DOPA or 5-HTP accumulated in the brain. The accumulation of DOPA was prevented by H 44/68 (methylester of α-methyl para-tyrosine), a tyrosine hydroxylase inhibitor. DOPA, which accumulated before H 44/68 was given, appeared stable for at least 20 min. There were no significant changes in the levels of NA, DA, 5-HT or tryptophan shortly after NSD 1015 administration, but a rise in tyrosine was noted. Increased brain tyrosine after l-tyrosine administration did not alter the DOPA accumulation, however. These data as well as the distribution of the accumulated amino acids suggest that the accumulation of DOPA and 5-HTP after decarboxylase inhibition occurs intraneuronally, that the decarboxylase enzyme is completely inhibited, and that the accumulated products are not appreciably metabolized or transported from the region studied. Amine synthesis rates and rate constants were calculated from the data and compare well with similar values determined by other methods. Thus this accumulation appears to be a reliable measure of the in vivo hydroxylation of tyrosine and tryptophan.