The activity of tyrosine hydroxylase in intact adrenergic neurons of the mouse vas deferens.
A procedure is described for measuring the activity of tyrosine hydroxylase in the intact adrenergic neurons of the mouse vas deferens. In this procedure, the L-dopa-1-14C which is formed from L-tyrosine-1-14C by the action of tyrosine hydroxylase is selectively and quantitatively decarboxylated by endogenous aromatic-L-amino acid decarboxylase. Although considerable tyrosine hydroxylase activity can be demonstrated with the intact mouse vas deferens in the absence of exogenous tetrahydropterin cofactor, the addition of 2-amino-4-hydroxy-6,7-dimethyltetrahydropteridine results in increased activity. Exogenous norepinephrine inhibits the activity of tyrosine hydroxylase in the intact mouse vas deferens and this inhibitory effect is competitively antagonized by 2-amino-4-hydroxy-6,7-dimethyltetrahydropteridine. When the vesicular catecholamine uptake mechanism is blocked by treatment of mice with 5 mg/kg of reserpine 24 hours prior to sacrifice, the activity of tyrosine hydroxylase in intact vas deferens is reduced and the inhibitory effect of exogenous norepinephrine is enhanced. Inhibition of monoamine oxidase with 0.15 mM pargyline does not affect the activity of tyrosine hydroxylase in the intact mouse vas deferens when the vesicular catecholamine uptake mechanism is intact but has a pronounced inhibitory effect following reserpine treatment. These observations lend further support to the conclusion that the activity of tyrosine hydroxylase in the intact adrenergic neuron is inversely related to the catecholamine concentration within an extravesicular pool. They also suggest that the catecholamines tend to accumulate within this extravesicular pool and thus become accessible to the action of monoamine oxidase when the vesicular uptake mechanism is inactivated or when the vesicular stores are filled to capacity.
D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D001681:Biological Assay; D001708:Biopterins / Q000494:pharmacology; D002250:Carbon Radioisotopes; D002395:Catecholamines / Q000032:analysis; D002396:Catechols / Q000032:analysis; D007700:Kinetics; D008297:Male; D008623:Mercaptoethanol / Q000494:pharmacology; D051379:Mice; D008995:Monoamine Oxidase / Q000502:physiology; D008996:Monoamine Oxidase Inhibitors; D009474:Neurons / Q000201:enzymology*; D009638:Norepinephrine / Q000032:analysis / Q000302:isolation & purification / Q000494:pharmacology; D010293:Pargyline / Q000494:pharmacology; D013564:Sympathetic Nervous System / Q000201:enzymology*; D014316:Tritium; D014443:Tyrosine / Q000494:pharmacology; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis / Q000378:metabolism*; D014649:Vas Deferens / Q000201:enzymology*
D002250:Carbon Radioisotopes; D002395:Catecholamines; D002396:Catechols; D008996:Monoamine Oxidase Inhibitors; D014316:Tritium; D001708:Biopterins; D014443:Tyrosine; D008623:Mercaptoethanol; D010293:Pargyline; D014446:Tyrosine 3-Monooxygenase; D008995:Monoamine Oxidase; D001205:Ascorbic Acid; D009638:Norepinephrine