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  • Title: Dopamine synthesis in synaptosomes: relation of autoreceptor functioning to pH, membrane depolarization, and intrasynaptosomal dopamine content.
    Author: Saller CF, Salama AI.
    Journal: J Neurochem; 1984 Sep; 43(3):675-88. PubMed ID: 6747629.
    Abstract:
    Factors affecting dopamine (DA) synthesis in rat striatal synaptosomes were examined by measuring the conversion of [3H]tyrosine (Tyr) to [3H]DA. Any [3H]DA that was synthesized was extracted into a toluene-based scintillation cocktail and quantitated by liquid scintillation spectrometry. The extraction was facilitated using di-(2-ethylhexyl) phosphoric acid (DEHP), a liquid cation exchanger. DA, apomorphine, and other DA agonists were much less potent inhibitors of DA synthesis in striatal synaptosomes at pH 6.2 than at pH 7.2. 3-(3-Hydroxyphenyl)-N-n-propylpiperidine (3-PPP), a putative DA autoreceptor agonist, was inactive at pH 6.2. However, at pH 7.2, 3-PPP did inhibit DA synthesis. This inhibition was reversed by sulpiride, a DA receptor antagonist, but not by benztropine, a DA uptake blocker, suggesting that 3-PPP inhibits DA synthesis by stimulating the DA autoreceptor. DA release from synaptosomes was much greater at pH 6.2 than at pH 7.2, most probably because the synaptosomal membrane appears to be depolarized at pH 6.2, as measured by the accumulation of [3H]tetraphenylphosphonium ions. Since tyrosine hydroxylase is inhibited by DA, this finding suggested that low assay buffer pH (i.e., pH 6.2) might interfere with the ability of 3-PPP and other DA agonists to inhibit DA synthesis, by promoting DA release. Likewise, reserpine and tetrabenazine, compounds which disrupt vesicular DA storage, were much less effective inhibitors of DA synthesis at pH 6.2 (high basal DA release). Moreover, D-amphetamine and high buffer potassium concentrations, treatments which promote DA release, also interfered with the ability of 3-PPP to inhibit DA synthesis. Thus, modulation of the release of DA in equilibrium with tyrosine hydroxylase may be a mechanism by which the DA autoreceptor regulates DA synthesis.
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