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Invited Symposium: MAOIs: Mulptiple Effects and Sites of Action






Abstract

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Discussion
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MAOIs: Interactions at Dopamine Receptors?


Contact Person: Beth Levant (blevant@kumc.edu)


[3H]Quinpirole and D2-like Receptors

Quinpirole (LY 171555) is a tricyclic ergoline and a relatively selective agonist at D2 and related dopamine receptors and has been extensively used in in vivo and in vitro studies of D2 receptor-mediated effects. Our laboratory and others have demonstrated specific, saturable, high affinity in vitro binding of [3H]quinpirole in rat striatal membranes and in autoradiographic studies which is appropriate for the classical D2 receptor with respect to pharmacological profile, guanine nucleotide regulation, and regional distribution of binding sites. Although not strictly identical to the binding profiles of other dopamine D2 receptor ligands, [3H]quinpirole binding was found to be consistent with what would be expected of dopamine D2 and closely related receptors with respect to pharmacological profile, guanine nucleotide and sodium regulation, and regional distribution in homogenate binding assays. As such, dopamine antagonists inhibited [3H]quinpirole binding with the following rank order of potencies: spiperone > (+)-butaclamol > haloperidol > (-)-sulpiride > clozapine > SCH 23390 >> cinanserin. The regional distribution of [3H]quinpirole binding sites was very similar to that of [3H]spiperone with very dense binding in the striatum, nucleus accumbens, and olfactory tubercles; moderate density in the substantia nigra/ventral tegmental area, pituitary, and hypothalamus; and low density in cortex and cerebellum. Some differentiation of dopamine D2 and D3 receptors in certain brain regions was apparent in autoradiographic studies using [3H]quinpirole. The overall pharmacological profile of the putative dopamine D2 and D3 receptors observed in autoradiographic studies was appropriate for dopamine D2-like receptors.

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MAOIs Inhibit [3H]Quinpirole Binding

The monoamine oxidase inhibitors (MAOIs) consist of a structurally diverse group of compounds whose antidepressant effects have been attributed to their common ability to inhibit monoamine oxidase (MAO) thus increasing the synaptic availability of one or more of the monoamine neurotransmitters. Surprisingly, we have shown that a variety of non-dopaminergic compounds, most-notably MAOIs, inhibit equilibrium [3H]quinpirole binding in rat striatal membranes in vitro (Table 1). In fact, clorgyline, Ro 41-1049, and phenelzine exhibited >1000-fold higher affinity in competition with [3H]quinpirole than [3H]spiperone. These observations suggest that MAOIs interact with the site labeled by [3H]quinpirole in brain. However, in keeping with the presumed lack of dopaminergic activity of most MAOIs, these compounds do not inhibit binding of other D2 receptor agonists or antagonists, such as [3H](-)-propylnorapomorphine and [3H]spiperone. This indicates that interaction with MAOIs is not simply a property of agonist binding at D2-like receptors and suggests that MQB may be unique to [3H]quinpirole, or perhaps the dopaminergic ergot derivatives.

Table 1. Potencies of monoamine oxidase-inhibitors at [3H]quinpirole-labeled
and [3H]spiperone-labeled sites in rat striatal membranes.
Compound/Ligand [3H]Quinpirole Ki [3H]Spiperone (nM)
Clorgyline 21 8 >50,000
Ro 41-1049 33 12 >100,000
Phenelzine 95 15 >100,000
Pargyline 85 6 >100,000
Tranylcypromine 269 45 6,633 1,293
(-)-Deprenyl 227 60 10,235 679
(+)-Deprenyl 589 56 >50,000
Isocarboxazid 2,160 241 >100,000
Nialamide 4,486 273 >100,000
Ro 16-6491 3,648 1,013 >100,000
Iproniazid 15,405 3,536 >100,000
Moclobemide 18,700 2,255 >100,000

Data represent the mean S.E. of 3 to 5 independent determinations as analyzed by LIGAND.

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MAO is not Involved in MQB

Although these structurally diverse compounds possess the common ability to inhibit the activity of MAO, several lines of evidence suggest that the enzymatic activity of MAO is not involved in this observation. First, the rank order of potencies of the MAOIs in competition for [3H]quinpirole binding does not correspond with the selectivities of these compounds for the MAO subtypes, A and B. For example, the MAO-A-selective drugs clorgyline and Ro 41-1049 were more than 400-fold more potent in inhibiting [3H]quinpirole binding than iproniazid which inhibits both MAO subtypes. Second, (+)-deprenyl, which is inactive at MAO, has nearly equal affinity to its active isomer (-)-deprenyl. Likewise, the interaction of these compounds with [3H]quinpirole binding is reversible while many of them, such as clorgyline, are irreversible inhibitors of MAO. In addition, quinpirole does not appear to be a substrate for MAO. Finally, these compounds inhibit [3H]quinpirole binding after steady-state has been reached. With the elimination of the involvement of MAO, interactions at the receptor level (e.g. an MQB site) must be considered.

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The MQB Site May be a D2-like Receptor

Although some MAOIs have been shown to inhibit binding of sigma, imidazoline, and dopamine uptake site ligands, the potency of MAOIs in competition for [3H]quinpirole binding correlates poorly with their potencies at these sites. Taken together, these data suggest that MAOIs may interact with a novel binding site in rat brain which is either labeled by [3H]quinpirole in addition to dopamine receptors or which modulates [3H]quinpirole binding at D2-like receptors. Subsequent study demonstrated that MQB in striatal membranes is not dependent on specific in vitro assay conditions, such as membrane concentration, the presence or absence of various ions, or incubation time or temperature . Finally, the distribution of MQB in rat brain regions and subcellular fractions parallels that of D2-like dopamine receptors. In fact, no data obtained to date suggests the binding of [3H]quinpirole to any site other than D2-like receptors. Thus, these observations suggest that MQB is associated with D2-like receptors.Further study must confirm whether MQB involves D2-like receptors. Even so, this observation has significant implications for the understanding of the pharmacology of MAOIs and quinpirole, and the nature of ligand-receptor interactions at D2-like receptors.Supported by NARSAD.

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References

  1. Levant, B., Grigoriadis, D.E., DeSouza, E.B. (1992) Characterization of [3H]quinpirole binding to D2-like dopamine receptors in rat brain. J. Pharmacol. Exp. Ther., 262:929-935.
  2. Levant, B., Grigoriadis, D.E., DeSouza, E.B. (1993) [3H]Quinpirole binding to putative D2 and D3 dopamine receptors in rat brain and pituitary gland: a quantitative autoradiographic study. J. Pharmacol. Exp. Ther., 264:991-1001.
  3. Levant, B., Grigoriadis, D.E., DeSouza, E.B. (1993) Monoamine oxidase inhibitors inhibit [3H]quinpirole binding in rat striatal membranes. Eur. J. Pharmacol., 246:171-178.
  4. Levant, B., Moehlenkamp, J.D., Morgan, K.A., Leonard, N.L., Cheng, C.C. (1996) Modulation of [3H]quinpirole binding in brain by monoamine oxidase inhibitors: evidence for a potential novel binding site. J. Pharmacol. Exp. Ther., 278:145-153.
  5. Fang, J., Yu, P. (1994) Effect of L-deprenyl, its structural analogues and some monoamine xoidase inhibitors on dopamine uptake. Neuropharmacology, 33:763-768.
  6. Itzhak, Y., Stein, I., Zhang, S.-H., Kassim, C.O., Cristante, D. (1991) Binding of sigma-ligands to C57BL/6 mouse brain membranes: effects of monoamine oxidase inhibitors and subcellular distribution studies suggest the existance of sigma-receptor subtypes. J. Pharmacol. Exp. Ther., 257:141-148.
  7. Olmos, G., Gabilondo, A.M., Miralles, A., Escriba, P.V., Garcia-Sevilla, J.A. (1993) Chronic treatment with the monoamine oxidase inhibitors clorgyline and pargyline down-regulates non-adrenoreceptor [3H]-idazoxan binding sites in the rat brain. Br. J. Pharmacol., 108:597-603.
  8. Levant B, Bancroft GN (1998) Inhibition of [3H]quinpirole binding by a monoamine oxidase inhibitor in subcellular fractions of rat striatum. Life Sci. 63:1643-1651.

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Levant, B; Moehlenkamp, JD; Bancroft, GN; Morgan, KA; (1998). MAOIs: Interactions at Dopamine Receptors?. Presented at INABIS '98 - 5th Internet World Congress on Biomedical Sciences at McMaster University, Canada, Dec 7-16th. Invited Symposium. Available at URL http://www.mcmaster.ca/inabis98/levant/levant0103/index.html
© 1998 Author(s) Hold Copyright