Estrogen rapidly potentiates amphetamine-induced striatal dopamine release and rotational behavior during microdialysis

Ovariectomy (OVX) of female rats results in a decreased behavioral response to stimulation of the mesostriatal dopamine (DA) system and decreased striatal DA release in vitro. Estrogen replacement restores both behavioral and neurochemical responsiveness. In this report, microdialysis in freely moving rats is used to simultaneously study the behavioral and neurochemical effects of systemic estradiol. OVX rats received a unilateral 6-hydroxy-dopamine lesion of the substantia nigra and then underwent microdialysis of the intact striatum. Thirty min after a single injection of 5 micrograms estradiol benzoate, amphetamine-induced rotational behavior and striatal DA release are both potentiated, relative to the response of oil-treated control animals.     

C-Fos identification of neuroanatomical sites associated with haloperidol and clozapine disruption of maternal behavior in the rat

Rat maternal behavior is a complex social behavior. Most antipsychotic drugs disrupt active maternal responses (e.g., pup retrieval, pup licking and nest building). Our previous work shows that typical antipsychotic haloperidol disrupts maternal behavior by blocking dopamine D₂ receptors, whereas atypical clozapine works by blocking 5-HT_2A/2C receptors. The present study used c-Fos immunohistochemistry technique, together with pharmacological tools and behavioral observations, and delineated the neuroanatomical bases of the disruptive effects of haloperidol and clozapine. Postpartum female rats were treated with haloperidol (0.2 mg/kg sc) or clozapine (10.0 mg/kg sc), with or without pretreatment of quinpirole (a selective dopamine D₂/D₃ agonist, 1.0 mg/kg sc) or 2,5-dimethoxy-4-iodo-amphetamine (DOI, a selective 5-HT_2A/2C agonist, 2.5 mg/kg sc). They were then sacrificed 2 h later after a maternal behavior test was conducted. Brain regions that have been previously implicated in the regulation of rat maternal behavior and/or in the antipsychotic action were examined. Behaviorally, both haloperidol and clozapine disrupted pup retrieval, pup licking and nest building. Pretreatment of quinpirole, but not DOI, reversed the haloperidol-induced disruptions. In contrast, pretreatment of DOI, but not quinpirole, reversed the clozapine-induced deficits. Neuroanatomically, the nucleus accumbens (both the shell and core), dorsolateral striatum and lateral septum showed increased c-Fos expression to the treatment of haloperidol. In contrast, the nucleus accumbens shell showed increased expression of c-Fos to the treatment of clozapine. More importantly, pretreatment of quinpirole and DOI produced opposite response profiles in the brain regions where haloperidol and clozapine had an effect. Based on these findings, we concluded that haloperidol disrupts active maternal behavior primarily by blocking dopamine D₂ receptors in a neural circuitry involving the nucleus accumbens, dorsolateral striatum and lateral septum. In contrast, clozapine appears to disrupt maternal behavior mainly by blocking serotonin 5-HT_2A/2C receptors in the nucleus accumbens shell.     

The influence of IgE-mediated reactions on the expression of delayed hypersensitivity in the rat.

The effect of IgE-mediated reactions on the expression of delayed hypersensitivity skin reactions was examined in the rat. It was found that the elicitation of an IgE-mediated reaction at the time of skin test could either potentiate or inhibit the development of delayed reactions. At a fixed level of IgE sensitivity, large delayed reactions were potentiated and small delayed reactions suppressed. These interactions were not dependent on the two types of sensitivity being direct against the same antigen but were dependent on the reaction of IgE with its appropriate antigen.     

Extracellular signal-regulated mitogen-activated protein kinase inhibitors decrease amphetamine-induced behavior and neuropeptide gene expression in the striatum

The aim of this study was to determine whether inhibition of the extracellular-regulated kinase signaling pathway decreases acute amphetamine-induced behavioral activity and neuropeptide gene expression in the rat striatum. Western blotting revealed that extracellular-regulated kinase1/2 phosphorylation was highly induced in the rat striatum 15 min after an acute amphetamine (2.5 mg/kg, i.p.) injection without altering the total amount of extracellular-regulated kinase protein. In a separate experiment, the systemic injection of SL327, a selective inhibitor of extracellular regulated kinase kinase that crosses the blood-brain barrier, 1 h prior to amphetamine administration decreased amphetamine-induced vertical and horizontal activity. Quantitative in situ hybridization histochemistry showed that SL327 abolished the high levels of preproenkephalin and preprodynorphin mRNA induced by amphetamine in the striatum with no alteration of their basal levels. In another set of experiments, the hyperlocomotor activity induced by amphetamine was reduced by pretreatment with intra-striatal infusion of U0126. U0126 also blocked the amphetamine-induced increases in phospho-extracellular-regulated kinase and preproenkephalin and preprodynorphin gene expression in the striatum. These data indicate that activation of the extracellular-regulated kinase cascade contributes to the behavioral effects and changes in striatal neuropeptide gene expression induced by acute amphetamine.     

The effects of cortical ablation on d-amphetamine-induced changes in striatal dopamine turnover and ascorbic acid catabolism in the rat.

Dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), ascorbic acid (AA) and dehydroascorbic acid (DHAA) levels were determined by HPLC in the striatal synaptosomal fraction and in the whole striatum of rats, whose fronto-parietal cortex had been bilaterally ablated, after a single injection of d-amphetamine (2.0 mg/kg i.p.). d-Amphetamine significantly increased the DHAA/AA ratio in unoperated and sham-operated rats, but failed to increase it in ablated rats, as compared to pertinent saline-treated groups. In the synaptosomal fraction, d-amphetamine significantly decreased the DHAA/AA ratio in unoperated, sham-operated and ablated rats. d-Amphetamine significantly decreased the DOPAC/DA ratio in the whole striatum and significantly increased it in the striatal synaptosomal fraction in all experimental groups. Cortical ablation greatly increased d-amphetamine-induced motor hyperactivity. We conclude that the d-amphetamine-induced increase in AA striatal oxidation requires integrity of the cortico-striatal glutamatergic pathways. Further, AA oxidation occurs in the extracellular space. The cortico-striatal glutamatergic pathways exert an inhibitory modulation on d-amphetamine behavioral effects.     

Melatonin attenuates the amphetamine-induced decrease in vesicular monoamine transporter-2 expression in postnatal rat striatum

The vesicular monoamine transporter-2 (VMAT-2) is responsible for packaging intraneuronal dopamine into synaptic vesicles in preparation for synaptic release and is a critical regulator of cytoplasmic dopamine levels and dopaminergic function. It has long been recognized that VMAT-2 is also a critical mediator of amphetamine-induced dopamine release. Amphetamine-induced lesions during development have the potential to produce numerous permanent abnormalities in neural circuitry and function. Therefore, in the present study, we investigated the effects of amphetamine on the levels of VMAT-2, α-synuclein and phosphorylated tyrosine hydroxylase in the striatum of neonatal rats. We found that chronic amphetamine administration in postnatal rats produces dopaminergic deficits in the striatum, including decreases in the levels of VMAT-2 and phosphorylated tyrosine hydroxylase. In addition, an increase in α-synuclein expression was observed in the striatum of postnatal rats following chronic amphetamine treatment. Furthermore, we identified a role of (10mg/kg) melatonin, a methoxyindole released from the pineal gland, in attenuating the detrimental effects of amphetamine on dopaminergic neurons.     

Differential effects of amphetamine and phencyclidine on the expression of growth-associated protein GAP-43

The purpose of the present study was to test changes in the expression of growth-associated protein (GAP-43) after chronic treatment with two different psychotomimetic drugs: amphetamine and phencyclidine. Rats were treated chronically for 7 days (twice daily) with 5 mg/kg of amphetamine and phencyclidine and sacrificed after 2, 5 or 7 days of treatment, and following 7, 14 or 21 days of recovery after full treatment (7 days). Separate groups of rats were treated on the same regiment with haloperidol, and control group was treated with vehicle. To determine the effects of different psychotomimetic drugs on the expression of GAP-43 we have used Northern blotting and quantitative in situ hybridization. Treatment with amphetamine induced decrease of GAP-43 mRNA expression, that was detected also during recovery period, up to 14 days after the last day of 7 days treatments. On the contrary, PCP induced increase of GAP-43 mRNA expression, that was detectable from the first days of treatment until 21 days after the last day of treatment. Treatment with haloperidol did not produce significant changes in GAP-43 mRNA expression. It can be suggested that GAP-43 upregulation upon phencyclidine treatment occurs as a result of functional activation of pathways able to participate in remodeling, while amphetamine showed neurotoxic effect, decreasing expression of GAP-43 mRNA.     

Excitatory effects of serotonin on rat striatal cholinergic interneurones

We investigated the effects of 5-hydroxytryptamine (5-HT, serotonin) in striatal cholinergic interneurones with gramicidin-perforated whole-cell patch recordings. Bath-application of serotonin (30 μ m ) significantly and reversibly increased the spontaneous firing rate of 37/45 cholinergic interneurones tested. On average, in the presence of serotonin, firing rate was 273 ± 193% of control. Selective agonists of 5-HT_1A, 5-HT₃, 5-HT₄ and 5-HT₇ receptors did not affect cholinergic interneurone firing, while the 5-HT₂ receptor agonist α-methyl-5-HT (30 μ m ) mimicked the excitatory effects of serotonin. Consistently, the 5-HT₂ receptor antagonist ketanserin (10 μ m ) fully blocked the excitatory effects of serotonin. Two prominent after-hyperpolarizations (AHPs), one of medium duration that was apamin-sensitive and followed individual spikes, and one that was slower and followed trains of spikes, were both strongly and reversibly reduced by serotonin; these effects were fully blocked by ketanserin. Conversely, the depolarizing sags observed during negative current injections and mediated by hyperpolarization-activated cationic currents were not affected. In the presence of apamin and tetrodotoxin, the slow AHP was strongly reduced by 5-HT, and fully abolished by the calcium channel blocker nickel. These results show that 5-HT exerts a powerful excitatory control on cholinergic interneurones via 5-HT₂ receptors, by suppressing the AHPs associated with two distinct calcium-activated potassium currents.     

Fluoxetine attenuates the effects of pentylenetetrazol on rat freezing behavior and c-Fos expression in the dorsomedial periaqueductal gray

The aim of the study was to investigate the role of the periaqueductal gray (PAG) in anxiolytic-like actions of fluoxetine in animals treated with an anxiogenic drug, pentylenetetrazol (PTZ), and subjected to fear conditioning procedure. The data showed that PTZ given at the dose of 30 mg/kg 15 min before a retention trial significantly decreased freezing reaction (p<0.01), and potently enhanced rat locomotor activity (p<0.01), in comparison to the control group. These effects were reversed by prior (60 min) administration of fluoxetine (20 mg/kg). Simultaneously, PTZ significantly increased c-Fos expression in the dorsomedial periaqueductal gray (DMPAG), examined 2h after the retention trial, in comparison to the control group (p<0.01). Fluoxetine (20 mg/kg) administered 60 min before PTZ reversed this effect. PTZ given at the same dose and time interval in the open field test did not affect rat locomotor behavior. Importantly, fluoxetine pretreatment did not change PTZ concentration in brain tissue. Our experiment based on PTZ-enhanced aversive conditioning revealed that acutely administered fluoxetine antagonized PTZ-induced panic-like behavior, and this phenomenon was accompanied by inhibition of activity of DMPAG.     

The effects of sensitization and social isolation on maternal behavior in the virgin rat

Thirty-three out of 67 virgin Sprague-Dawley rats spontaneously retrieved 5–10-day-old foster pups in a pretest. The 34 rats not responding immediately were randomly divided into three groups to study the effects of direct and exteroceptive exposure to young on the induction of maternal behavior. Retrieving, licking and crouching behavior appeared in a higher proportion of cage rats (n = 12) which received a fresh group of pups to live with throughout each day than in control rats (n = 10) that had only 5 min daily exposure. However basket rats (n = 12) also became maternal merely by exposure to exteroceptive stimuli from pups living in baskets fixed to the cage wall. In a second experiment, virgin rats that spontaneously retrieved pups were socially isolated for 2–4 weeks. As a result, a 50–60% decrease in the number of rats showing retrieval was found not only in an independent sample of rats (n = 20) but also in a related group (n = 13) that practiced the response in the week between a pretest and Week 2 in isolation. Therefore, the results demonstrate that exteroceptive cues from pups alone may be sufficient to induce maternal behavior in nonspontaneously retrieving virgin rats, and that isolation from pups and other social stimuli reduce retrieving behavior in initially responsive virgin rats.     

Local infusion of interleukin-6 attenuates the neurotoxic effects of NMDA on rat striatal cholinergic neurons.

The potential neuroprotective effects of IL-6 against the excitotoxic neuronal loss induced by N-methyl-D-aspartate (NMDA) have been studied. Infusion into the rat striatum of excitotoxic amounts (250 nmol) of NMDA resulted in a 45% decrease in striatal choline acetyl transferase activity (ChAT; a marker of cholinergic neurons) and glutamate decarboxylase (GAD, a marker of GABAergic neurons) at 2 days post-injection. Co-infusion of 10 U of IL-6 reduced the loss of ChAT activity to 21% but failed to prevent the loss of GAD activity. IL-6 per se, up to the dose of 500 U, failed to affect ChAT or GAD activities. The in vivo effects of IL-6 are not mediated by a direct antagonism of NMDA toxicity, since IL-6 (up to a concentration of 500 and 5000 U/ml, respectively) did not antagonize either the increase in cyclic GMP levels resulting from NMDA receptor activation in cerebellar slices or the glutamate-induced release of lactate dehydrogenase, an index of neurotoxicity, by cultured cortical neurons. These results suggest that the increase in IL-6 levels observed in experimental brain lesions may play a role in the protection and regeneration of cholinergic neurons.     

Acute fluoxetine administration differentially affects brain C-Fos expression in fasted and refed rats

In the present study we investigated the effect of acute fluoxetine administration on the expression of c-Fos in the rat brain under two different metabolic conditions: fed and fasting states. Wistar male rats, weighing 220+/-30g, received i.p. injections of saline solution or fluoxetine (10mg/kg), and were killed 2 h later. The brains were removed after transcardiac perfusion with phosphate-buffered saline followed by paraformaldehyde, and were then processed for immunohistochemistry. Fos-like immunoreactivity was quantified by a computerized system. Fasted animals faced an 18-h suppression of food intake, while fed groups were submitted to an initial 14-h period of fast followed by a 4-h period in which food was freely available. Both in fasting and fed states, fluoxetine-treated animals presented a significant increase in c-Fos expression in hypothalamic areas, limbic structures, circumventricular areas, and in mesencephalic and rhomboencephalic regions, as compared with saline-treated controls. The quantitative comparison of data obtained from fasted and fed animals showed that fasted rats treated with fluoxetine presented a higher c-Fos expression in the ventromedial hypothalamus and the paraventricular nuclei compared with the fed group, while in fluoxetine-treated fed rats c-Fos expression was higher in the arcuate nuclei, medial amygdala, locus coeruleus and dorsal raphe nuclei, as compared with fasted, fluoxetine-treated animals. These data indicate that the metabolic condition of the animals significantly modifies fluoxetine-induced brain c-Fos expression, suggesting that visceral and behavioral fluoxetine effects may be influenced by the metabolic state of the individual.     

The effects of sexual experience and estrus on male-seeking motivated behavior in the female rat

Ovariectomized (OVX) female rats were trained to traverse a straight alley and return to a goal box where they had previously encountered a male rat, a female rat or an empty goal box. The time required to run the alley was used as an index of the subjects' motivation to re-engage the goal box target. Subjects were tested in both estrus and non-estrus, first sexually naïve and then again after sexual experience. Female rats ran most quickly for a male target, most slowly for an empty goal box, and at intermediate speeds for a female target. Sexual experience tended to slow run times for all but male targets. Estrus enhanced approach behavior for males and an empty goal box, but tended to slow the approach toward females, both before and after sexual experience. This latter finding was further investigated in a second experiment in which sexually naïve OVX females were tested during estrus and non-estrus in a locomotor activity apparatus, a runway with an empty goal box, and an open field. Estrus produced no changes in spontaneous locomotion either in the activity box or the open field, but decreased run times in the alley and increased the number of center-square entries in the open-field. Thus, estrus produces increases in sexual motivation that selectively enhance exploratory, presumably male-seeking behavior, but not simple spontaneous locomotion.     

Yohimbine and naloxone: effects on male rat sexual behavior.

We evaluated the effects of yohimbine (2 mg/kg) and naloxone (5 mg/kg), separately and in combination, on copulatory behavior in male rats. In Experiment 1, yohimbine evinced decrements in intromission frequency, ejaculation latency, and copulatory efficiency, whereas naloxone administration was followed by an increased ejaculation latency, and the combination of yohimbine plus naloxone was without effect. In Experiment 2, yohimbine evinced decreases in intromission frequency, ejaculation latency, copulatory efficiency in the first, but not subsequent, copulatory series, as well as a decreased latency to sexual exhaustion. Further, treatment with yohimbine alone, naloxone alone, or yohimbine plus naloxone was followed by a reduction in the number of ejaculation prior to sexual exhaustion. Thus, at the doses tested, no synergistic effects were observed for the combination of yohimbine plus naloxone.     

Cortical stimulation induces fos expression in striatal neurons.

Electrical stimulation of a broad area of the frontoparietal cortex in the rat brain induces immunocytochemically detectable Fos in striatal neurons normally devoid of the protein. The vividness of labeling within striatal neurons was maximal at 0.5 h after the cessation of a 15-min-long stimulation period and became weaker by 3 h. Although Fos-reactive neurons were widely distributed in the striata of both hemispheres in an uneven pattern, those on the stimulated side were more numerous and more darkly stained. At no time-point were labeled neurons found in the globus pallidus, entopeduncular nucleus or substantia nigra. Destruction of the nigrostriatal dopamine projection with 6-hydroxydopamine did not induce Fos production and failed to prevent the induction of Fos by cortical stimulation. That many of the Fos-positive neurons project to the substantia nigra was confirmed by retrograde labeling with Fluoro-Gold. The data suggest that corticostriatal excitatory transmission may directly influence the genomic activity of striatal neurons by way of Fos.     

Early and delayed biochemical effects of paraquat toxicity on rat lung

This research was undertaken to study the early and delayed biochemical effects of paraquat dichloride on the lungs of rats. To accomplish this, female rats were given a sublethal dose (18 mg/kg) of paraquat, in a single (ip) injection. At the end of 24 hr, only catalase, β-N-acetylglucosaminidase, and glucose-6-phosphate dehydrogenase (G6PD) activities and nonprotein sulfhydryl (NPSH) levels were significantly (P < 0.05) increased above saline (0.9%) -injected controls. However, at 6 days after paraquat injection, all biochemical indices measured were significantly (P < 0.02) above those values of controls. These findings indicate that the biochemical changes produced by paraquat might be consistent with the changes in the population of cell types, which are well documented histologically by other investigators.     

Short-term plasticity at inhibitory synapses in rat striatum and its effects on striatal output

Two forms of short-term plasticity at inhibitory synapses were investigated in adult rat striatal brain slices using intracellular recordings. Intrastriatal stimulation in the presence of the ionotropic glutamate receptor antagonists 6-cyano-7-nitroquinoxaline-2,3-dione (20 microM) and D,L-2-amino-5-phosphonovaleric acid (50 microM) produced an inhibitory postsynaptic potential (IPSP) that reversed polarity at -76 +/- 1 (SE) mV and was sensitive to bicuculline (30 microM). The IPSP rectified at hyperpolarized membrane potentials due in part to activation of K(+) channels. The IPSP exhibited two forms of short-term plasticity, paired-pulse depression (PPD) and synaptic augmentation. PPD lasted for several seconds and was greatest at interstimulus intervals (ISIs) of several hundred milliseconds, reducing the IPSP to 80 +/- 2% of its control amplitude at an ISI of 200 ms. Augmentation of the IPSP, elicited by a conditioning train of 15 stimuli applied at 20 Hz, was 119 +/- 1% of control when sampled 2 s after the conditioning train. Augmentation decayed with a time constant of 10 s. We tested if PPD and augmentation modify the ability of the IPSP to prevent the generation of action potentials. A train of action potentials triggered by a depolarizing current injection of constant amplitude could be interrupted by stimulation of an IPSP. If this IPSP was the second in a pair of IPSPs, it was less effective in blocking spikes due to PPD. By contrast, augmented IPSPs were more effective in blocking spikes. The same results were achieved when action potentials were triggered by a depolarizing current injection of varying amplitude, a manipulation that produces nearly identical spike times from trial to trial and approximates the in vivo behavior of these neurons. These results demonstrate that short-term plasticity of inhibition can modify the output of the striatum and thus may be an important component of information processing during behaviors that involve the striatum.     

Effect of dopamine-depleting brain lesions in rat pups: role of striatal serotonergic neurons in behavior

Previous results from our laboratory have demonstrated that 3-day-old rats given dopamine (DA)-depleting brain lesions are spared the severe behavioral dysfunctions seen after comparable brain damage in adults. This behavioral sparing is accompanied by a sprouting of serotonin (5-HT)-containing neurons in the striatum. The present results extend these observations by demonstrating that rats given the brain lesions as 15- or 27-day-olds continue to suckle, wean, and grow into adulthood without exhibiting any obvious behavioral dysfunctions, yet striatal 5-HT levels do not increase. Moreover, combined destruction of DA- and 5-HT-containing neurons in 3-day-old rat pups also produced no obvious behavioral dysfunctions. These and other results indicate that increases in striatal 5-HT are not necessary for the behavioral sparing observed after DA-depleting brain lesions in neonatal rats.