Differential action of bromocriptine on nigrostriatal versus mesolimbic dopaminergic neurons

Summary The present study was undertaken to compare the abilities of the dopaminergic agonists apomorphine, bromocriptine, and lergotrile to inhibit the synthesis of dopamine (DA) in terminals of nigrostriatal and mesolimbic DA neurons. Thein vivo synthesis of DA was estimated by measuring the rate of accumulation of dihydroxyphenylalanine (DOPA) in terminals of nigrostriatal (striatum) and mesolimbic (nucleus accumbens, olfactory tubercle) neurons 30 min after the administration of NSD 1015, a decarboxylase inhibitor. The activation of DA autoreceptors in these regions was evaluated by measuring the abilities of the DA agonists to inhibit DA synthesis in brain regions of rats pretreated with gamma-butyrolactone (GBL). Apomorphine (0.03–1.0 mg/kg for 45 min) and bromocriptine (0.1–10 mg/kg for 90 min) produced dose-dependent decreases in the rate of DA synthesis in all three brain regions of both vehicle- and GBL-treated rats. A time course of the effects of the highest dose of bromocriptine (10 mg/kg), however, demonstrated dramatic regional differences in the ability of this drug to inhibit DA synthesis in saline-versus GBL-pretreated rats. Bromocriptine inhibited the GBL-induced increase in DA synthesis for 6 hours in all regions examined. In the striatum of saline-treated rats the decrease in DA synthesis was evident only at 1.5 hours after bromocriptine administration, while in the nucleus accumbens and olfactory tubercle DA synthesis remained inhibited for 6 hours. By contrast, lergotrile reduced DA synthesis to a similar extent in all three regions for at least 6 hours in both vehicle- and GBL-treated rats. These results suggest that there is no regional difference in the ability of bromocriptine to inhibit DA synthesis via DA autoreceptor mechanisms, but there appear to be differences in post-synaptic DA receptor-mediated mechanisms which regulate nigrostriatal versus mesolimbic DA neurons.     

Autoradiographic evidence for nicotine receptors on nigrostriatal and mesolimbic dopaminergic neurons

Rats received unilateral injections of 6-hydroxydopamine into the medial forebrain bundle, resulting in an ipsilateral loss of striatal dopamine and of dopaminergic perikarya. A concimitant reduction of displaceable tritiated nicotine binding was observed autoradiographically in the ipsilateral substantia nigra, ventral tegmental area, striatum, nucleus accumbens, and olfactory tubercle. Thus, nicotine receptors appear to be located on nigrostriatal and mesolimbic dopaminergic neurons at the level of perikarya and terminals.     

Intra-accumbens implants of embryonic dopaminergic neurons reverse the behavioral supersensitivity to opiates evoked by lesion of the mesolimbic dopaminergic pathway

The ascending mesotelencephalic dopaminergic systems of rat pups of 3 days of age were bilaterally lesioned using 6-hydroxydopamine injected at the level of the lateral hypothalamus. A sub-group of lesioned pups received, 5 days after the lesion, a dopamine neuron-rich cell suspension graft implanted bilaterally into the striatum and nucleus accumbens. Behavioral tests were conducted 6 months later. The lesion induced an increase in the locomotor activation induced by D-Ala2-Met5-enkephalinamide injected into the nucleus accumbens (2.5 micrograms/side) as compared to the activation observed in control animals. Locomotor activation by systemic apomorphine (0.1 mg/kg s.c.) was also increased while that induced by amphetamine (1.5 mg/kg i.p.) was abolished. The presence of DA neuron implants reversed each of these post-lesion modifications.     

Failure to detect collateral sprouting of mesolimbic dopaminergic neurons during early postnatal development

Using biochemical parameters the present study sought to assess the normal developmental pattern of the dopaminergic innervation of the olfactory tubercle (OT) and how it is affected by olfactory bulbectomy. In rats, the adult pattern of cellular organization is achieved in the OT gradually over the first 7 days after birth. On the other hand, tyrosine hydroxylase (TH) and [3H]dopamine ([3H]DA) uptake, while present at low levels, start to increase rapidly only after the first 7 days reaching adult levels by 40 and 20 days after birth, respectively. TH in dopamine (DA) cell bodies of A10 was already high, 40% of adult value, at birth, reached 150% by day 14 and decreased back to adult values by day 21 after birth. In 10-day-old rats, bulbectomy resulted, 30 days later, in an increase to 123% of control in TH activity and 137% in [3H]DA uptake within the OT. Comparable changes were found following bulbectomy in adults. However, bulbectomy in 1-day-old rats did not produce any significant changes 40 days later. The findings suggest that during postnatal growth TH activity is increased in DA cell bodies, preceding the changes in DA terminals of the OT, resembling the changes occurring during collateral sprouting in adults. In addition, changes indicative of collateral sprouting do not occur in response to deafferentation of the OT in 1-day-olds but do in 10-day-olds or older animals, a phenomenon probably related to a critical development period of the OT.     

Autoreceptor-mediated changes in dopaminergic terminal excitability: Effects of striatal drug infusions

The neurophysiological correlates of autoinhibition at the terminals of nigrostriatal dopaminergic neurons were studied by measuring the changes in antidromic excitability of nigrostriatal neurons following local infusions of various catecholamine agonists and antagonists into the neostriatum. Infusions of apomorphine or amphetamine reduced terminal excitability whereas the dopamine antagonists, haloperidol, fluphenazine or sulpiride, led to increases in terminal excitability. Alterations in antidromic excitability were constrained to the terminal regions and were not observed when infusions and excitability testing were performed in the medial forebrain bundle. The alpha-2 agonist, clonidine, did not alter dopaminergic terminal excitability. Our results indicate that pharmacological manipulations which have been shown to reduce the amount of stimulation-induced transmitter release from dopaminergic terminals are associated with a dopamine autoreceptor-mediated hyperpolarization and/or alteration in ionic conductance of the terminal membranes. These results are discussed with respect to mechanisms of autoinhibition in the central nervous system.     

Autoreceptor-mediated changes in dopaminergic terminal excitability: Effects of potassium channel blockers

The effects of the potassium channel blockers, 4-aminopyridine (4-AP) and tetraethylammonium (TEA), on autoreceptor-mediated changes in dopaminergic terminal excitability were examined in urethane-anesthetized rats. Local infusions of 4-AP or TEA into neostriatal terminal fields of nigral dopaminergic neurons led to marked decreases in terminal excitability, as measured by the increase in stimulating current required to activate the neurons antidromically from the site of the infusion. The decreased excitability resulting from 4-AP could be reversed by subsequent i.v. injection of haloperidol, and was blocked in rats that had been depleted of endogenous dopamine by prior treatment with alpha-methyl-p-tyrosine (AMpT). Thus, the decrease in excitability elicited by the potassium channel-blockers was indirect, and apparently due to increased autoreceptor stimulation resulting from enhanced transmitter release. In addition, co-infusion of 4-AP and apomorphine in AMpT-treated animals led to decreased terminal excitability that did not differ from the effects of apomorphine alone, indicating that 4-AP did not block the effects of exogenous autoreceptor agonist administration. These results provide in situ electrophysiological evidence that autoreceptor-mediated processes occurring at dopaminergic terminals are not mediated by 4-AP- or TEA-sensitive potassium channels. Furthermore, our findings suggest that, as in other types of presynaptic terminals, blockade of voltage-sensitive potassium channels in dopamine terminals leads to enhanced release of transmitter.     

Activation of striatal dopaminergic grafts by haloperidol

The effect of haloperidol (0.5 mg/kg, 60 min) on striatal dopamine metabolism of intact rats and of rats bearing a dopaminergic graft implanted into the previously denervated striatum was investigated. The dopaminergic grafts increased dopamine and dihydroxyphenylacetic acetic acid (DOPAC) contents to 6.2 and 9.6% of their respective control levels (lesioned striatum: 0.5 and 0.7% respectively). Haloperidol increased both the DOPAC content and the DOPAC:DA ratio in the grafted striatum, and the magnitude of these increases were similar to those seen in intact controls. Furthermore a tendency for the high striatal DOPAC:DA ratio seen in lesioned striata to revert toward control values could also be observed in grafted animals. The significance of these results is discussed in terms of the in vivo regulation of graft activity.     

Autoreceptor-mediated changes in dopaminergic terminal excitability: Effects of increases in impulse flow

In frogs poisoned with botulinum toxin type A the quantal content of endplate potentials is greatly reduced. Lowering the temperature of the preparation increases quantum content; between 14 and 4 degrees C the mean Q10 for this effect is 6.3. Facilitation of synaptic transmission is marked with pairs of stimuli and cooling further enhances facilitation. The time constant of decay of facilitation is 34 ms at 20 degrees C and 116 ms at 4 degrees C. The increase in facilitation and in its time constant of decay at low temperature are presumably not a result of a prolongation of the duration of the nerve terminal action potential since such changes are not seen in the presence of K+-channel blockade by 3,4-diaminopyridine. Electrotonic depolarization of nerve terminals in the presence of tetrodotoxin and 3,4-diaminopyridine induces all-or-none endplate currents. Such endplate currents, at a holding potential of -50 mV, show that the amount of charge entry is about 1/3 of that in unpoisoned junctions but still corresponds to 5-10 X 10(3) transmitter quanta. Transmitter release at this level is maintained during repetitive stimulation even in the presence of 82 mM Ca2+ in the extracellular solution. We speculate that the blockade of transmitter release in BoTx -poisoned muscles results from a stimulatory effect of the toxin on metabolic systems of Ca2+ disposal in the nerve terminal.     

Decoding of dopaminergic mesolimbic activity and depressive behavior

Dopaminergic mesolimbic and mesocortical systems are involved in hedonia and motivation, two core symptoms of depression. However, their role in the pathophysiology of depression and their manipulation to treat depression has received little attention. Previously, we showed decreased limbic dopamine (DA) neurotransmission in an animal model of depression, Flinder sensitive line (FSL) rats. Here we describe a high correlation between phase–space algorithm of bursting-like activity of DA cells in the ventral tegmental area (VTA) and efficiency of DA release in the accumbens. This bursting-like activity of VTA DA cells of FSL rats is characterized by a low dimension complexity. Treatment with the antidepressant desipramine affected both the dimension complexity of cell firing in the VTA and rate of DA release in the accumbens, as well as alleviating depressive-like behavior. Our findings indicate the potential usefulness of monitoring limbic dopaminergic dynamics in combination with non-linear analysis. Decoding the functionality of the dopaminergic system may help in development of future antidepressant drugs.     

Dopaminergic terminal excitability following arrival of the nerve impulse: The influence of amphetamine and haloperidol

Variations in the excitability of the axons and terminal fields of dopaminergic neurons of the substantia nigra were examined as a function of time following the nerve impulse in urethane-anesthetized, immobilized rats. Excitability was measured by determining the threshold, defined as the minimum current required to consistently activate dopaminergic neurons antidromically. Threshold currents were maximal immediately following the action potential and declined exponentially to a plateau. The interval during which threshold current declined, the phasic period, was significantly longer for stimulation of neostriatal terminal fields as contrasted to stimulation of axons along the medial forebrain bundle. A positive correlation was observed between antidromic response latency and the duration of this phasic period for both sites of stimulation, an observation consistent with the view that the site of initiation of the antidromic action potential is of smaller diameter in the neostriatum than in the medial forebrain bundle Amphetamine, which promotes dopamine release and/or re-uptake blockade, reduced dopaminergic terminal excitability in the neostriatum at all intervals examined. This effect increased at successively shorter intervals during the phasic portion of the excitability curve. Haloperidol, a dopamine antagonist, increased the excitability of dopaminergic terminal fields, an effect which was also more marked earlier in the phasic interval. Neither amphetamine nor haloperidol had a significant effect on the excitability of dopaminergic axons in the medial forebrain bundle. Variations in dopaminergic terminal excitability after impulse arrival, and the effects of amphetamine and haloperidol on this phenomenon suggest that terminal excitability is determined by events related to arrival of the nerve impulse including activation of presynaptic 'autoreceptors' by dopamine released from the synaptic ending.     

Reversible reduction of tyrosine hydroxylase enzyme protein during the retrograde reaction in mesolimbic dopaminergic neurons

Changes in the activity and amount of the neurotransmitter synthesizing enzyme tyrosine hydroxylase (TH) were measured in dopaminergic (DA) neurons of the A10-mesolimbic system of the rat following electrolytic lesions of their axons. Unilateral hypothalamic lesions in close proximity to the cell bodies resulted, within 24--48 h, in a permanent anterograde reduction of TH to 10--20% of control in the ipsilateral olfactory tubercle and nucleus accumbens. The retrograde reaction in the A10-DA nerve cell bodies was characterized by an initial increase in TH activity to 133% by 24--48 h followed by a gradual and permanent fall to 50% of control by day 14 due to retrograde cell death of DA neurons. In contrast, lesions of DA axon terminals in the olfactory tubercle resulted in a reversible retrograde reduction of TH activity of the A10. The enzyme activity declined during the first 7 days to 70% of control and then gradually recovered, reaching control levels by 28 days after the operation. The reduction in TH activity in the A10 was demonstrated by immunochemical titration with a specific antibody to TH to be entirely due to reduced amounts of enzyme protein. We conclude that in mesolimbic DA neurons; (a) the anterograde reaction is characterized by a rapid and permanent decline of TH in degenerating terminals; (b) the retrograde reaction is dependent upon the proximity of the lesion to the nerve cell body, and (c) a reversible reduced accumulation of TH characterizes the retrograde reaction in response to lesions of distal axons. The reaction of DA neurons of the mesolimbic system to axonal injury is comparable to that of the nigrostriatal system.     

Increased neuronal responsiveness to cholecystokinin and dopamine induced by lesioning mesolimbic dopaminergic neurons: an electrophysiological study in the rat

In the rat, cholecystokinin (CCK) and dopamine (DA) coexist in a subpopulation of neurons of the ventral tegmental area (VTA) projecting to the nucleus accumbens. However, in the dorsal hippocampus, dopaminergic projections from the VTA do not contain CCK, the latter neurotransmitter being mainly localized in intrinsic hippocampal neurons. The present experiments were undertaken in order to compare the interactions of CCK and DA and the effects of lesioning VTA dopaminergic neurons in a region where these neurotransmitters coexist and in one where they do not. The effects of microiontophoretic applications of CCK, kainate (KA), glutamate (GLU) and DA were determined in control rats and in rats pretreated with a local injection of 6-hydroxydopamine (6-OHDA) in the VTA. In the nucleus accumbens and in the hippocampus of intact rats, DA exerted a similar depressant effect whether applied during CCK-, KA- or GLU-induced activations. The 6-OHDA lesion enhanced responsiveness of accumbens neurons to KA, GLU and CCK (the responsiveness to this latter peptide being increased by more than 15-fold) and the depressant effect of DA when applied during neuronal activation by KA or GLU but not when the same neurons were activated with CCK. In the dorsal hippocampus, the 6-OHDA lesion enhanced neuronal responsiveness to KA and DA in the CA1, but not in the CA3 region, whereas the responsiveness to CCK remained unchanged in both regions. These results suggest a physiological role for the coexistence of CCK and DA in the nucleus accumbens. The induction of a supersensitivity to DA in the CA1, but not in the CA3, region of the dorsal hippocampus following a VTA lesion is consistent with the regional distribution of the dopaminergic innervation in this structure.     

Mesocortical dopaminergic neurons. 2. Electrophysiological consequences of terminal autoreceptor activation

Measurement of drug- and stimulation-induced changes in the electrical excitability of dopaminergic terminals was employed to assess the effects of stimulation of dopamine terminal autoreceptors in the prefrontal cortex in urethane-anesthetized rats. Systemic or local administration of amphetamine decreased, whereas systemic administration of haloperidol increased the excitability of prefrontal cortical dopaminergic terminals of ventral tegmental area dopaminergic neurons. Mesoprefrontal dopaminergic terminal excitability was also responsive to spontaneous and stimulation-induced alterations in the rate of impulses reaching the terminal fields. These results are comparable to those previously reported for nigrostriatal and mesoaccumbens dopaminergic neurons, and are discussed with regard to the operational characteristics of autoinhibition in the mesocortical dopaminergic system.     

Modulation of the rat mesolimbic dopamine pathway by neurokinins.

The locomotor activity (LMA) response induced after infusion of selective neurokinin (NK) agonists into the cell body (A10) and a terminal region of the mesolimbic pathway of the rat was investigated. Infusion of the NK1 receptor-selective agonist, GR73632, into the ventral tegmental area (VTA: A10) or the nucleus accumbens (NAS) significantly and dose-dependently increased basal LMA. Agonists selective for the NK2 and NK3 receptors, GR64349 and senktide respectively, had no effect on LMA after intra-NAS infusion. The LMA induced by GR73632 is mediated via dopamine (DA) since the response was abolished by haloperidol. From these studies it would appear that the elevated LMA reported previously after VTA or NAS administration of substance P probably occurs via NK1 receptors. Such data supports the notion that endogenous NKs are likely to be important in modulating the mesolimbic DA pathway and, as a consequence, compounds which antagonise their effects could be useful for the treatment of disorders associated with this system. However, simultaneous infusion of the NK1 agonists, +/- CP-96,345 and its analogue CPQ, into the VTA did not attenuate the LMA induced after intra-VTA infusion of GR73632. Co-infusion of the NK1 antagonist CPQ, but not +/- CP-96,345, attenuated the LMA response induced by GR73632 in the NAS. The apparent poor susceptibility of these responses to blockade by the recently developed non-peptide NK1 antagonists was unexpected but may reflect their poor affinity for the rat variant of the NK1 receptor.(ABSTRACT TRUNCATED AT 250 WORDS)     

Modulation of hippocampal excitability and seizures by galanin.

Previous studies have shown that the expression of the neuropeptide galanin in the hippocampus is altered by seizures and that exogenous administration of galanin into the hippocampus attenuates seizure severity. To address the role of endogenous galanin in modulation of hippocampal excitability and its possible role in seizure mechanisms, we studied two types of transgenic mice: mice with a targeted disruption of the galanin gene (GalKO) and mice that overexpress the galanin gene under a dopamine-beta-hydroxylase promoter (GalOE). GalKO mice showed increased propensity to develop status epilepticus after perforant path stimulation or systemic kainic acid, as well as greater severity of pentylenetetrazol-induced convulsions. By contrast, GalOE mice had increased resistance to seizure induction in all three models. Physiological tests of hippocampal excitability revealed enhanced perforant path-dentate gyrus long-term potentiation (LTP) in GalKO and reduced LTP in GalOE. GalKO showed increased duration of afterdischarge (AD) evoked from the dentate gyrus by perforant path simulation, whereas GalOE had increased threshold for AD induction. Depolarization-induced glutamate release from hippocampal slices was greater in GalKO and lower in GalOE, suggesting that alterations of physiological and seizure responses in galanin transgenic animals may be mediated through modulation of glutamate release. Our data provide further evidence that hippocampal galanin acts as an endogenous anticonvulsant and suggest that genetically induced changes in galanin expression modulate both hippocampal excitability and predisposition to epileptic seizures.     

Modulation of H-reflex excitability by tetanic stimulation.

OBJECTIVE: This study investigated a strategy to elicit reversible facilitation of the soleus monosynaptic H-reflex in humans using a modified tetanic stimulation technique. METHODS: Interventional tetanic stimulation (ITS) was applied to the tibial nerve in the popliteal fossa, and soleus H-reflexes were recorded before and after stimulation in 15 healthy volunteers. RESULTS: ITS resulted in significantly increased soleus H-reflex amplitudes that outlasted the stimulation period by approximately 16 min. The effect of ITS on soleus motor evoked potentials to transcranial magnetic stimulation and on somatosensory evoked potentials to tibial nerve stimulation was also investigated; no significant changes were found. CONCLUSIONS: ITS produced a reversible increase in H-reflexes in the absence of changes in motor evoked potential or somatosensory evoked potential that outlasted the intervention period for up to 16 min. SIGNIFICANCE: This technique may be used in future studies to investigate whether the induced increased H-reflex excitability influences locomotion.     

Modulation of long-term depression by dopamine in the mesolimbic system.

Long-lasting adaptations in the mesolimbic dopamine (DA) system in response to drugs of abuse likely mediate many of the behavioral changes that underlie addiction. Recent work suggests that long-term changes in synaptic strength at excitatory synapses in the two major components of this system, the nucleus accumbens (NAc) and ventral tegmental area, may be particularly important for the development of drug-induced sensitization, a process that may contribute to addiction, as well as for normal response-reinforcement learning. Using whole-cell patch-clamp recording techniques from in vitro slice preparations, we have examined the existence and basic mechanisms of long-term depression (LTD) at excitatory synapses on both GABAergic medium spiny neurons in the NAc and dopaminergic neurons in the midbrain. We find that both sets of synapses express LTD but that their basic triggering mechanisms differ. Furthermore, DA blocks the induction of LTD in the midbrain via activation of D2-like receptors but has minimal effects on LTD in the NAc. The existence of LTD in mesolimbic structures and its modulation by DA represent mechanisms that may contribute to the modifications of neural circuitry that mediate reward-related learning as well as the development of addiction.