Acute and chronic methylphenidate administration in intact and VTA-specific and nonspecific lesioned rats.

Methylphenidate (MPD) is a psychostimulant used for the treatment of ADHD and works by increasing the bioavailability of dopamine (DA) in the brain. As a major source of DA, the ventral tegmental area (VTA) served as the principal target in this study as we aimed to understand its role in modulating the acute and chronic MPD effect. Forty-eight male Sprague-Dawley rats were divided into control, sham, electrical lesion, and 6-OHDA lesion groups. Given the VTA's implication in the locomotive circuit, three locomotor indices-horizontal activity, number of stereotypy, and total distance-were used to measure the animals' behavioral response to the drug. Baseline recording was obtained on experimental day 1 (ED 1) followed by surgery on ED 2. After recovery, the behavioral recordings were resumed on ED 8. All groups received daily intraperitoneal injections of 2.5 mg/kg MPD for six days after which the animals received no treatment for 3 days. On ED 18, 2.5 mg/kg MPD was re-administered to assess for the chronic effect of the psychostimulant. Except for one index, there was an increase in locomotive activity in all experimental groups after surgery (in comparison to baseline activity), acute MPD exposure, induction with six daily doses, and after MPD re-challenge. Furthermore, the increase was greatest in the electrical VTA lesion group and lowest in the 6-OHDA VTA lesion group. In conclusion, the results of this study suggest that the VTA may not be the primary nucleus of MPD action, and the VTA plays an inhibitory role in the locomotive circuit.

Reinforcement omission effects in rats with bilateral lesions in the substantia nigra pars compacta and ventral tegmental area

Reinforcement omission effects (ROEs) are characterized by higher response rates after reinforcement omission than after reinforcement delivery. This pattern of behavior is interpreted in terms of motivational and attentional processes. Recent studies from our laboratory have shown that the amygdala, nucleus accumbens, and medial prefrontal cortex are involved in ROE modulation. Also, the literature has demonstrated a role of other areas such as substantia nigra pars compacta (SNc) and the ventral tegmental area (VTA) in processes related to surprising events, such as prediction error and presentation or omission of an event (exteroceptive stimulus and reinforcement). Since these structures send projections to areas related to ROE modulation such as the amygdala, nucleus accumbens, and prefrontal cortex, the objective of the present study was to determine whether the SNc and VTA also integrate the circuit involved in ROE modulation. Rats were trained on a fixed-interval 12 s with limited-hold 6 s signaled schedule of reinforcement (Pre-lesion training). After acquisition of stable performance, the rats received bilateral neurotoxic lesions of the SNc (Experiment 1) and VTA (Experiment 2). Following postoperative recovery, the rats were submitted to two refresher sessions (Post-lesion training). Subsequently, the training was changed from a 100 to a 50% schedule of reinforcement (Post-lesion testing). In both experiments, the results showed that there was no difference in performance between sham rats and rats with bilateral lesions of the SNc or the VTA.

Dopaminergic projections from midbrain to primary motor cortex mediate motor skill learning.

The primary motor cortex (M1) of the rat contains dopaminergic terminals. The origin of this dopaminergic projection and its functional role for movement are obscure. Other areas of cortex receive dopaminergic projections from the ventral tegmental area (VTA) of the midbrain, and these projections are involved in learning phenomena. We therefore hypothesized that M1 receives a dopaminergic projection from VTA and that this projection mediates the learning of a motor skill by inducing cellular plasticity events in M1. Retrograde tracing from M1 of Long-Evans rats in conjunction with tyrosine hydroxylase immunohistochemistry identified dopaminergic cell bodies in VTA. Electrical stimulation of VTA induced expression of the immediate-early gene c-fos in M1, which was blocked by intracortical injections of D(1) and D(2) antagonists. Destroying VTA dopaminergic neurons prevented the improvements in forelimb reaching seen in controls during daily training. Learning recovered on administration of levodopa into the M1 of VTA-lesioned animals. Lesioning VTA did not affect performance of an already learned skill, hence, left movement execution intact. These findings provide evidence that dopaminergic terminals in M1 originate in VTA, contribute to M1 plasticity, and are necessary for successful motor skill learning. Because VTA dopaminergic neurons are known to signal rewards, the VTA-to-M1 projection is a candidate for relaying reward information that could directly support the encoding of a motor skill within M1.

Lesion of the ventral tegmental area amplifies stimulation-induced Fos expression in the rat brain.

Unilateral lesions of the ventral tegmental area (VTA), the key structure of the mesolimbic system, facilitate behavioral responses induced by electrical stimulation of the VTA in the contralateral hemisphere. In search of the neuronal mechanism behind this phenomenon, Fos expression was used to measure neuronal activation of the target mesolimbic structures in rats subjected to unilateral electrocoagulation and simultaneously to contralateral electrical stimulation of the VTA (L/S group). These were compared to the level of mesolimbic activation after unilateral electrocoagulation of the VTA (L group), unilateral electrical stimulation of the VTA (S group) and bilateral electrode implantation into the VTA in the sham (Sh) group. We found that unilateral stimulation of the VTA alone increased the density of Fos containing neurons in the ipsilateral mesolimbic target structures: nucleus accumbens, lateral septum and amygdala in comparison with the sham group. However, unilateral lesion of the VTA was devoid of effect in non-stimulated (L) rats and it significantly amplified the stimulation-induced Fos-immunoreactivity (L/S vs S group). Stimulation of the VTA performed after contralateral lesion (L/S) evoked strong bilateral induction of Fos expression in the mesolimbic structures involved in motivation and reward (nucleus accumbens and lateral septum) and the processing of the reinforcing properties of olfactory stimuli (anterior cortical amygdaloid nucleus) in parallel with facilitation of behavioral function measured as shortened latency of eating or exploration. Our data suggest that VTA lesion sensitizes mesolimbic system to stimuli by suppressing an inhibitory influence of brain areas afferenting the VTA.

6-hydroxydopamine lesions enhance progesterone-facilitated lordosis of rats and hamsters, independent of effects on motor behavior.

The Ventral Tegmental Area (VTA) is an important brain area for progesterone (P(4))'s effects to facilitate female sexual behavior of rodents. We investigated the importance of dopaminergic neurons in the VTA, and two dopaminergic projection sites, the Nucleus Accumbens (NAc), and Caudate Nucleus of the Striatum (CN), in modulating P(4)-facilitated sex and motor behavior. Ovariectomized (ovx) rats and hamsters, administered estradiol benzoate (10 microg) and P(4) (0, 50, 100, 200, or 500 microg), were tested for motor behavior in a chamber that automatically records horizontal beam breaks, and for sexual behavior in response to a sexually-experienced male. Animals were tested once a week until each P(4) dosage was received; animals then had bilateral 6-hydroxydopamine (6-OHDA) or sham lesions to the VTA, NAc, or CN and were re-tested at each P(4) dosage on subsequent weeks. Fixed brains were stained with cresyl violet and processed for dopamine transporter (DAT) immunoreactivity. The number of cresyl violet stained cells was significantly lower in all 6-OHDA infusion sites compared to non-6-OHDA infusion sites of rats and hamsters. Also, in rats, the number of DAT-immunoreactive neurons was lower in all 6-OHDA infusion sites compared to non-6-OHDA infusion sites. In rats, 6-OHDA but not sham, lesions to the VTA, NAc, or CN produced P(4)-dependent increases in lordosis quotients and resulted in modest increases in motor behavior. In hamsters, 6-OHDA, but not sham, lesions to the VTA, NAc, or CN produced P(4)-dependent increases in total lordosis durations and produced modest decreases in motor behavior. This suggests that the dopaminergic output neurons of midbrain VTA may play an important role in modulation of P(4)-facilitated sexual lordosis among rodents.

Quantitative analyses of GFRalpha-1 and GFRalpha-2 mRNAs and tyrosine hydroxylase protein in the nigrostriatal system reveal bilateral compensatory changes following unilateral 6-OHDA lesions in the rat.

Copy numbers of mRNAs for GFRalpha-1 and GFRalpha-2, the preferred receptors for glial cell line-derived neurotrophic factor (GDNF) and neurturin (NTN) were determined by real-time quantitative RT-PCR (QRT-PCR). Receptor expression was assessed in striatum (ST) and substantia nigra (SN) of normal rats and rats acutely or progressively lesioned by 6-OHDA injected into the medial forebrain bundle or ST, respectively. GFRalpha-1 mRNA was clearly detected in normal ST. In normal SN, significantly higher expression of both receptors was observed. At 4 weeks after acute lesion, GFRalpha-2 mRNA was markedly decreased in SN bilaterally, whereas GFRalpha-1 mRNA in SN and ST was not affected. A progressive lesion resulted in a progressive decrease of GFRalpha1 mRNA in ST bilaterally. In SN, levels of GFRalpha-1 mRNA were not significantly affected by a progressive lesion, whereas GFRalpha-2 mRNA was markedly decreased bilaterally. Quantitative western blotting standardized against tyrosine hydroxylase (TH) protein from PC12 cells revealed the expected decrease in TH protein in lesioned SN, but also significant increases in TH protein in contralateral, unlesioned SNs at 4 weeks after both acute and progressive lesions. These data suggest that previously unrecognized compensatory changes in the nigrostriatal system occur in response to unilateral dopamine depletion. Since the changes observed in receptor expression did not always parallel loss of dopamine neurons, cells in addition to the nigral dopamine neurons appear to be affected by a 6-OHDA insult and are potential targets for the neurotrophic factors, GDNF and NTN.