ABSTRACT
The role of catecholamine neuronal input on GABAergic activity in the hypothalamus, telencephalon, optic tectum, and cerebellum was investigated in early recrudescent female goldfish (Carassius auratus). A new quantitative technique was developed and validated, permitting concomitant quantification and correlational analysis of glutamic acid decarboxylase 65 (GAD65), GAD67, and GAD3 mRNA levels and in vivo GABA synthesis. Catecholamine depletion was achieved by the administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP; 50 microg/g body weight) and dopamine (DA) depletion verified by HPLC. Endogenous GABA levels were increased by intraperitoneal administration of gamma-vinyl GABA (GVG; 300 microg/g body weight), an inhibitor of the GABA catabolic enzyme GABA transaminase. Treatment with MPTP resulted in a greater than twofold increase in GABA synthesis rate in the optic tectum and telencephalon. The increase in GABA synthesis rate was highly correlated with an increase in GAD67, but not GAD65 or GAD3 mRNA levels. These results suggest that catecholaminergic input exerts inhibitory effects on GABA synthesis rates through the modulation of GAD67 in the optic tectum and telencephalon. Together with previously published observations in rodents and primates, it is suggested that catecholaminergic control of GABA synthesis must have evolved more than 200 million years ago, before the emergence of the teleost fishes.
Subject(s)
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine/pharmacology , Brain/drug effects , Catecholamines/deficiency , Dopamine Agents/pharmacology , Glutamate Decarboxylase/metabolism , Isoenzymes/metabolism , gamma-Aminobutyric Acid/metabolism , Analysis of Variance , Animals , Brain/anatomy & histology , Brain/enzymology , Brain Chemistry/drug effects , Chromatography, High Pressure Liquid/methods , Dopamine/metabolism , Electrophoretic Mobility Shift Assay/methods , Enzyme Inhibitors/pharmacology , Glutamate Decarboxylase/genetics , Goldfish , Isoenzymes/genetics , RNA, Messenger/drug effects , RNA, Messenger/metabolism , Time Factors , Tissue Distribution , Tritium/pharmacokinetics , Vigabatrin/pharmacologyABSTRACT
Antifungal activity is positively correlated to furanocoumarin content in extracts of the traditional phytomedicine northern prickly ash (Zanthoxylum americanum Mill. [Rutaceae]). The specificity of these furanocoumarins in inhibiting replication of DNA was investigated with reference to significant base composition differences between fungal and mammalian mitochondrial DNA. We developed a polymerase chain reaction-based assay to investigate whether (1) furanocoumarins inhibit DNA polymerization and (2) distinct furanocoumarins specifically inhibit DNA replication depending on base composition. Specific inhibition of DNA polymerization by 5-methoxypsoralen and psoralen through high-adenine and thymine (AT) (84.3%) and low-AT (51.9%) DNA, respectively, suggests that furanocoumarins inhibit replicative functions of genomes or of regions within the genome that differ in base composition. Greater overall inhibition of DNA polymerization by Z. americanum husk extracts than with single or mixed furanocoumarins suggests that inhibitory compounds in addition to the major furanocoumarins are present in Z. americanum.
