André Ferron

Modulatory role for biogenic amines in the cerebral cortex. Microiontophoretic studies

In order to investigate the mode of action of biogenic amines in rat cerebral cortex, the unitary activity of spontaneously firing neurons and their excitatory response to acetylcholine (ACh) were examined using microiontophoretic administration of dopamine (DA), noradrenaline (NA) and serotonin (5-HT). The predominant effect of these biogenic amines on the spontaneous activity was a profound and prolonged inhibition of firing (2-4 min), which attained its maximum within 15-120 sec. This response was generally more abrupt in onset and of greater magnitude with NA and 5-HT than with DA. Most units inhibited by DA, NA and 5-HT also showed marked depression of their excitatory response to ACh when pretreated with these biogenic amines. With repetitive administration of ACh, it could be shown that the total duration of inhibition of ACh responses by DA and NA was not as prolonged as the inhibition of the spontaneous firing of the same cells. With 5-HT, the initial ACh responses of many neurons could be completely blocked, and this inhibitory effect lasted as long as the inhibition of spontaneous firing. In view of the anatomical data demonstrating a relative sparsity of monoamine nerve terminals in cerebral cortex, the strong inhibition induced by DA, NA or 5-HT may have reflected slow inactivation of the biogenic amines. However, it could also be indicative of underlying mechanisms of action dependent on metabolic changes. Indeed, the interaction between biogenic amines and ACh might imply a balance between the intracellular pools of cAMP and cGMP is directly or indirectly influenced by the biogenic amines and ACh, respectively. This hypothesis would not exclude other modes of local interaction between DA, NA, 5-HT and ACh, and appears compatible with the modulatory role of biogenic amines in cerebral cortex.

Changes of D1 and D2 receptors in adult rat neostriatum after neonatal dopamine denervation: Quantitative data from ligand binding,in situ hybridization and iontophoresis

The specific binding of [3H]SCH23390 to D1 and of [3H]raclopride to D2 dopamine receptors was measured by autoradiography in the rostral and caudal halves of neostriatum and in the substantia nigra of adult rats subjected to near total destruction of nigrostriatal dopamine neurons by intraventricular 6-hydroxydopamine soon after birth. Three months after this lesion, [3H]SCH23390 binding (D1 receptors) was slightly but significantly decreased in the rostral neostriatum (22%), but unchanged in its caudal half and in the substantia nigra. In contrast, [3H]raclopride binding (D2 receptors) was considerably increased throughout the neostriatum (10-40%), while markedly decreased in the substantia nigra (80%). In the rostral neostriatum, there were no parallel changes in D2 receptor messenger RNA levels, as measured by in situ hybridization on adjacent sections. Caudally, however, slight but significant increases in D2 messenger RNA could be observed (10-20%). As assessed by quantitative iontophoresis, there was a marked enhancement (63%) of the inhibitory responsiveness of spontaneously firing units in the rostral neostriatum to dopamine and the D1 agonist, SKF38393, in neonatally lesioned compared to control rats. On the other hand, responsiveness to PPHT, a potent D2 agonist, appeared to be unchanged. Such opposite changes in the number of D1 and D2 binding sites, dissociated from the expression of D2 receptor messenger RNA and from the sensitivity to dopamine and D1 and D2 agonists, suggested independent adaptations of these various parameters following the neonatal dopamine denervation of neostriatum. They also provided further evidence for mechanisms other than the dopamine innervation in the control of the expression of neostriatal D2 receptor messenger RNA during ontogenesis, and emphasized that the effects of dopamine and its D1 and D2 agonists in neostriatum do not depend strictly on the number of D1 and D2 primary ligand recognition sites.

Effects ofp-chlorophenylalanine on cortical monoamines and on the activity of noradrenergic neurons

The catecholamines noradrenaline, dopamine, adrenaline, the indoleamine 5-hydroxy-tryptamine (5-HT; serotonin), and some of their major metabolites were assayed, using high performance liquid chromatography (HPLC), in the neocortex of normal rats as well as in animals in which 5-HT synthesis had been inhibited withp-chlorophenylalanine. Besides important depletions in serotonin and in 5-hydroxyindole-3-acetic acid, noradrenaline levels were significantly reduced, but the content in 3-methoxy-4-hydroxyphenylglycol was increased, indicating an augmented utilization of this amine. The levels of dopamine and 3-methoxytyramine were also reduced, although homovanillic acid and 3,4-dihydroxyphenylacetic acid levels remained constant. The spontaneous unitary activity of identified noradrenergic neurons in the Locus coeruleus was increased, indicating an hyperactivity of this system. These results can be interpreted in relation to functional interactions between the catecholamines and serotonin; i.e.: a decrease in endogenous serotonin results in the loss of a negative feedback control of noradrenaline release.

Hypersensitivity to serotonin and its agonists in serotonin-hyperinnervated neostriatum after neonatal dopamine denervation

Neonatal destruction of the nigrostriatal dopamine projection by intraventricular 6-hydroxydopamine leads to a serotonin (5-hydroxytryptamine, 5-HT) hyperinnervation of the adult neostriatum accompanied by increased radioligand binding to 5-HT1B, 5-HT1nonAB and 5-HT2 receptors. The consequences of such 5-HT receptor changes on neuronal responsiveness to 5-HT and corresponding receptor agonists were assessed with a quantitative iontophoretic approach. For comparative purposes, similar data were also obtained from rats 6-hydroxydopamine lesioned as adults, showing severe neostriatal dopamine denervation but no 5-HT hyperinnervation. In controls, 5-HT and its receptor agonists, m-chlorophenylpiperazine (mCPP; 5-HT1B/2C agonist) and dimethoxy-iodophenyl-aminopropane (DOI; 5-HT2A/2C agonist), depressed the firing rate of a majority of the unit tested. Three months after neonatal 6-hydroxydopamine lesion (5-HT-hyperinnervated tissue), inhibitory responses to all three agents were significantly increased and comparable results were obtained for 5-HT and DOI in the rostral versus caudal neostriatum. After 6-hydroxydopamine lesion in adults, neither responsiveness to 5-HT, mCPP or DOI nor the density of 5-HT1B or 5-HT2A binding were significantly different from control. Thus, the up-regulation of 5-HT1B, 5-HT2A and possibly 5-HT2C receptors accompanying the 5-HT hyperinnervation after neonatal but not after adult dopamine denervation was associated with increased responsiveness (IT50) of neostriatal neurons to iontophoresed 5-HT and its receptor agonists. Under these conditions, neostriatal 5-HT transmission might be enhanced in spite of a basal release seemingly comparable to normal (Jackson and Abercrombie, 1992, J. Neurochem. 58, 890).(ABSTRACT TRUNCATED AT 250 WORDS)

Dopamine modulates the inhibition induced by GABA in rat cerebral cortex: an iontophoretic study

Effects of iontophoresed gamma-aminobutyric acid (GABA) and two GABA agonists, 4,5,6,7-tetrahydroisooxazolo-[5,4-c]pyridine-3-ol (THIP) and baclofen were quantitatively compared in the anterior cingulate, frontal, and parietal cortex of urethane-anesthetized intact rats after catecholamine (CA) depletion with alpha-methyl-p-tyrosine (alpha-MPT) or selective dopamine (DA) denervation with 6-hydroxydopamine (6-OHDA). As assessed with to the IT50 index, the postsynaptic sensitivity to GABA was significantly higher in anterior cingulate than in frontal and parietal cortex. The responsiveness to GABA was also greater in frontal than in parietal cortex. Sensitivity to GABA was significantly reduced in both anterior cingulate and frontal cortex after CA depletion, and similarly, after DA denervation with 6-OHDA. The difference in the sensitivity to GABA between the three cortical regions in intact rats as well as after CA depletion did not seem to be correlated with either GABAA or GABAB receptors since the responsiveness to both GABA agonists in every region examined was comparable in intact rats, and remained unchanged after alpha-MPT treatment. This finding raises the possibility that some GABA receptors in the cerebral cortex may be pharmacologically distinct from the two main subtypes of GABA receptors, GABAA and GABAB. When GABA was administered by iontophoresis in the anterior cingulate cortex during continuous applications of subthreshold currents of DA, the inhibition induced by GABA was either increased or decreased. As DA innervation density is nearly two-fold greater in anterior cingulate than in frontal cortex, and 30-fold greater in anterior cingulate than in parietal cortex, these results suggest that responsiveness to GABA may be correlated with the regional density of DA innervation and that elevated levels of DA may enhance the sensitivity to GABA.

Electrophysiological effects of methylphenidate on the coeruleo-cortical noradrenergic system in the rat

The effect of methylphenidate on noradrenergic neurotransmission was investigated in urethane-anesthetized rats. The spontaneous activity of locus coeruleus noradrenergic neurons was the same in rats treated for 7 days with methylphenidate as in the controls. In control rats, i.v. methylphenidate induced a reduction of locus coeruleus neuronal firing whereas in rats treated for 7 days with methylphenidate, the same dose of methylphenidate failed to induce any change in locus coeruleus activity. At this time, clonidine induced a lesser reduction of locus coeruleus neuronal firing than in the controls, indicating that their autoreceptors had become desensitized. Following electrical stimulation of the locus coeruleus, most of the spontaneously firing cortical neurons were inhibited but the percentage of such neurons was reduced and the neurons showed a decreased responsiveness after methylphenidate treatment. The responsiveness of cortical neurons to microiontophoretic applications of NA as assessed by the I.T50 method was reduced after 7 days of treatment with methylphenidate. These findings suggest that the efficacy of cortical NA neurotransmission is markedly reduced following methylphenidate treatment.

Possible existence of a presynaptic positive feedback mechanism enhancing dopamine transmission in the anterior cingulate cortex of the rat

A series of microiontophoretic and VTA stimulation experiments, conducted in intact, GBR-12909-treated, α-methylparatyrosine-depleted or 6-hydroxydopamine-denervated rats, provide suggestive evidence for the existence of a presynaptic, positive feedback mechanism triggered by dopamine reuptake and favoring the release of this transmitter in the anterior cingulate cortex.

Acute and chronic effects of methylphenidate on cortical adrenoceptors in the rat

The effects of acute and chronic treatments with methylphenidate were examined on cortical adrenoceptors. A single dose of methylphenidate increased the affinity of [3H]prazosin and [3H]idazoxan binding sites. Acute and chronic methylphenidate treatments caused a down-regulation of alpha 1-adrenoceptors. There were no changes in cortical beta-adrenoceptors measured with [3H]dihydroalprenolol nor in endogenous monoamine levels. The alterations in alpha-adrenoceptors can be attributed to an indirect action of methylphenidate possibly through an accrued release of noradrenaline or tyramine.

The Heterogeneity of the Catecholamine Innervation of Cerebral Cortex

The central catecholamine (CA) neurons in the brain stem and their CNS projections were the first to be described in correlative biochemical and histofluorescent investigations as chemically identified neurotransmitter systems. By combining histofluorescent, autoradiographic, and immunocytochemical methods with biochemical determinations of endogenous levels and activities of the enzymes of synthesis, three types of CA neuronal systems were described in the CNS, according to the monoamine they can synthesize and presumably use as their neurotransmitter, i.e., dopamine (DA; Berger et al, 1974; Hokfelt et al, 1974; Lindvall et al, 1974), noradrenaline (NA; Carlsson et al., 1962; Dahlstrom and Fuxe, 1964; Fuxe, 1965; Fuxe et al., 1968; Ungerstedt, 1971; Lindvall and Bjorklund, 1974), and adrenaline (AD; Hokfelt et al, 1973; Van der Gugten et al, 1976; Versteeg et al, 1976; Reader, 1981).