Dopamine-Sensitive Adenylate Cyclase in Mammalian Brain: A Possible Site of Action of Antipsychotic Drugs

Clement‐Cormier, Yvonne C.; Kebabian, John W.; Petzold, Gary L.; Greengard, Paul

doi:10.1073/pnas.71.4.1113

Cited by 383 publications

(69 citation statements)

References 12 publications

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“…This suggests that, in addition to anticholinergic action, these drugs have a hitherto unsuspected inhibitory effect on the cyclase which tends to be reversed by the dopamine synthesized from L-dopa. These parallelisms break down, however, when one considers that anticholinergics, which are hallucinogens, have acted in the opposite direction in tests such as these (4)(5)(6)24).…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Opposing effects of dopaminergic to cholinergic compounds on a cerebral dopamine-activated adenylate cyclase.

Tang¹,

Cotzias²

1977

Proc. Natl. Acad. Sci. U.S.A.

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We have studied these possibilities in the hope of advancing the treatments of Parkinson's disease and of psychosis. To this end we used adenylate cyclase from mouse caudate to screen agents that can affect the symptoms of Parkinson's disease. These included drugs that stimulate the dopaminergic system in man or animals and inactive metabolites and precursors thereof and compounds that activate or inhibit the cholinergic system, which counterbalances the dopaminergic one in the brain (7,8).MATERIALS AND METHODS Animals. These were 5-to 6-week-old male Swiss albino mice, weighing 23-27 g, maintained on Purina Chow and water ad lib., and kept in an air-conditioned room on a 12-hr lightto-darkness cycle. The experiments were started in midday by decapitating the animals in a cold room. The brains were removed within 30 sec and placed on a cold watch glass. The lateral ventricles were entered by removing their roofs. The exposed heads of the caudates were scooped out with forceps.Determination of Dopamine-Stimulated Adenylate Cyclase Activity. The methods are essentially those developed in Greengard's laboratory (9). All fragments of both caudates (about 30 mg) were placed in Krebs-Ringer bicarbonate solution, pH 7.4, which had been gassed for 2 min with 5% CO2/ 95% 02. They were homogenized in 0.5 ml of 50 mM Trismaleate buffer, pH 7.4, containing 12.5 mM theophylline, 0.25 mM ethylene glycol-bis(Q-aminoethyl ether)-NN'-tetraacetic acid, and 2.5 mM MgSO4. Aliquots (0.4 ml) of the homogenates were placed in 5-ml Pyrex tubes that had previously been equilibrated to 30°in an orbital water bath. The control tubes received 50 gl of H20. The experimental tubes received different additives in different series of experiments, the concentrations of which are given below. In one series, 50 ,ul containing either dopamine or a dopaminergic substance was added. In another series, the tubesi received 25 ,ul containing one of the above substances plus 25 jul containing-an inhibitor, a metabolite, or a precursor thereof. In another series, the precursors and metabolites were tested in various concentrations for activation of the cyclase. In still another series, the metabolites were screened for inhibition of the dopamine-dependent activation. Finally, some cholinergic and anticholinergic agents were screened for activation or inhibition of the cyclase.The reaction was started by adding 50 jul of a 0.5 mM solution of ATP with mixing and then incubating in a 30°orbital water bath. After 2.5 min, the reaction was terminated by immersion of the tubes into a boiling water bath for 2 min. The homogenates were centrifuged at 1500 X g for 5 min. The supernatant solutions were quick-frozen until analyzed for adenosine 3': 5'-cyclic monophosphate (cyclic AMP) according to Gilman (10).The results of the cyclic AMP determinations were used as follows. We first computed the mean ±SEM of both nonstimulated and stimulated samples. From these we calculated the net dopamine-stimulated cyclic AMP produced by subtracting the mean nonstimulated acti...

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Section: Resultsmentioning

confidence: 99%

“…The (4)(5)(6). Because this cyclase is cardinally involved in the receipt and amplification of messages by postsynaptic neurons at dopaminergic synapses, it was possible that dopaminergic antiparkinsonism agents may function by activating this cyclase.…”

mentioning

confidence: 99%

Opposing effects of dopaminergic to cholinergic compounds on a cerebral dopamine-activated adenylate cyclase.

Tang¹,

Cotzias²

1977

Proc. Natl. Acad. Sci. U.S.A.

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“…For example, clinically effective antipsychotic agents displace dopamine ligands from striatal binding sites (Creese et al, 1976); they block dopamine-stimulated adenylate cyclase in preparations of caudate homogenates (Clement-Cormier et al, 1974;Miller et al, 1974); and they increase striatal concentrations of DA metabolites (Bacopoulos and Roth, 198 1;Carlsson and Lindquist, 1953). In behavioral paradigms, systemically administered apomorphine antagonizes neuroleptic-induced catalepsy (Kolbe et al,198 1).…”

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confidence: 99%

Investigation of the failure of parenterally administered haloperidol to antagonize dopamine released from micropipettes in the caudate

Johnson¹,

Hoffer²,

Freedman³

1986

J. Neurosci.

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An anomaly in the experimental data underlying the theory that neuroleptics act by blockade of dopaminergic neurotransmission is the repeatedly demonstrated failure in several laboratories of parenterally administered neuroleptics to antagonize electrophysiologic actions of locally applied dopamine (DA) in striaturn. This failure is enigmatic since many investigators have successfully demonstrated antagonism when both dopamine and neuroleptic are applied directly to striatal neurons by microiontophoresis. We used multibarrel micropipettes to pressure-eject DA agonists onto rat caudate neurons while observing the ability of parenterally administered haloperidol to block the inhibitory actions of dopaminergic agonists on neuronal activity. Experiments performed at times of maximal behavioral effect of haloperidol did not demonstrate agonist-antagonist interaction. This result has been obtained by four other teams of investigators. A variety of pharmacologic manipulations were employed to help solve this enigma. Acute treatment with reserpine and a-methyl-paratyrosine, performed to minimize any possible interference by endogenous DA, did not permit blockade of dopamine by haloperidol. To see if this failure of antagonism could be generalized to other DA agonists, apomorphine, amphetamine, and phencyclidine (PCP) were also investigated. Although the direct dopaminergic agonist apomorphine was not antagonized by haloperidol, the indirect DA agonist PCP was successfully antagonized. Amphetamine, which has both direct and indirect actions when applied locally, was not antagonized. Antagonism of direct agonists was demonstrated in rats with unilateral 6-hydroxydopamine-induced lesions of the nigrostriatal pathway. In these preparations, parenterally administered haloperidol reversed the receptor-mediated supersensitivity to the inhibitory effects of locally applied DA and apomorphine. These data suggest that in caudate there exist postsynaptic receptors that are more sensitive to haloperidol than those receptors usually activated by DA released into the neuronal milieu from micropipettes. We suggest that PCP's indirect action on endogenous DA in presynaptic terminals results in specific interaction with these receptors. We further suggest that the postsynaptic receptors most sensitive to blockade by haloperidol may also be those receptors that increase in number following denervation. Failure to demonstrate antagonism of direct DA agonists in the absence of supersensitivity suggests that locally applied DA does not normally contact postsynaptic dopami-

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“…Effects on dopamine receptors determine actions of major psychotropic drugs (1). Antagonism by neuroleptic "antischizophrenic" drugs of a dopamine-sensitive adenylate cyclase correlates with pharmacological potency, suggesting that the cyclase is associated with the dopamine receptor (2)(3)(4)(5). We now report direct binding of dopamine to apparent postsynaptic receptor sites in membrane fractions of the corpus striatum of calf and rat.…”

mentioning

confidence: 99%

Dopamine receptor binding in the corpus striatum of mammalian brain.

Burt¹,

Enna²,

Creese³

et al. 1975

Proc. Natl. Acad. Sci. U.S.A.

187

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METHODSCaudate or other regions of calf brains, fresh from a slaughterhouse, were frozen at -200 until assayed (up to 2 weeks). Tissue was homogenized with a Brinkmann Polytron PT-10 in 40 volumes of ice-cold 50 mM Tris-HCI buffer, pH 7.7 at 250. The homogenate was centrifuged at 50,000 X g for 10 min on a Sorvall RC-2B centrifuge. The supernatant fluid was discarded; the pellet was rehomogenized in 50 volumes of the same buffer and recentrifuged. The supernatant fluid was discarded, and the pellet was homogenized in 100 volumes of 50 mM Tris-HCI buffer, pH 7.5 at 250, with 10 uM pargyline and 0.1% (weight/volume) ascorbic acid. This final tissue resuspension was placed in a 370 bath for 5 min and returned to ice for use in the binding assay.The two preliminary washes with buffer remove endogenous dopamine, while preincubation at 370 ensures inactivation of tissue monoamine oxidase by pargyline before addition of radioactive dopamine.Male Sprague-Dawley rats were decapitated and the corpus striatum was prepared as described for frozen calf tissue.[3H]Dopamine (3,4-dihydroxyphenyl[ethyl-1-3H

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Dopamine-Sensitive Adenylate Cyclase in Mammalian Brain: A Possible Site of Action of Antipsychotic Drugs

Cited by 383 publications

References 12 publications

Opposing effects of dopaminergic to cholinergic compounds on a cerebral dopamine-activated adenylate cyclase.

Opposing effects of dopaminergic to cholinergic compounds on a cerebral dopamine-activated adenylate cyclase.

Investigation of the failure of parenterally administered haloperidol to antagonize dopamine released from micropipettes in the caudate

Dopamine receptor binding in the corpus striatum of mammalian brain.

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