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Aims: This study aims to investigate whether 3,4-dihydroxyphenylacetic acid (DHAA) can improve gut barrier function by inhibiting the mitogen-activated protein kinase (MAPK) - myosin light-chain kinase (MLCK) signaling pathway in type 2 diabetes (T2D) mice., Main Methods: T2D mice were induced by a high-fat diet combined with streptozotocin. T2D mice were intragastrically administered with DHAA at 75 and 150 mg kg/body weight per day for 4 weeks. Blood glucose, insulin level, oxidative stress and inflammatory cytokines were measured. TJ (tight junction) protein and MAPK-MLCK pathway-related proteins were analyzed by western blot., Key Findings: DHAA alleviated hyperglycemia and decreased insulin resistance of T2D mice. It also decreased oxidative stress via increased glutathione (GSH) and total superoxide dismutase (T-SOD) activities and reduced containing malondialdehyde (MDA). DHAA exhibited a significant anti-inflammatory effect by decreasing the level of pro-inflammatory cytokines lipopolysaccharide (LPS) and interleukin (IL)-6 and increasing that of anti-inflammatory cytokine IL-10. More importantly, DHAA improved gut barrier function by enhancing tight junction protein expression and inhibiting the MAPK-MLCK signaling pathway., Significance: DHAA could reduce oxidative stress, decrease inflammatory response, and improve intestinal function in T2D mice, which may help to relieve the symptoms of T2D., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Aims: To investigate dynamic changes and roles of melatonin (MLT) in the striata of 6-hydroxydopamine (6-OHDA)-treated rats., Main Methods: A Parkinson's disease (PD) rat was established by a unilateral injection of 6-OHDA into the right substantia nigra pars compacta (SNc) and the right medial forebrain bundle (MFB) to achieve a complete lesion of the ipsilateral nigrostriatal DA system. Dialysates were collected in the lesioned striatum at different time intervals by in vivo microdialysis. In addition, both contralateral and ipsilateral striatum tissues were collected at two time intervals (10:00 and 22:00 h) at 3 and 6 weeks after lesioning. The levels of DA, 3, 4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the dialysates, as well as MLT in the dialysates and tissues were determined using HPLC., Key Findings: The dialysate contents of DA, DOPAC and HVA in the lesioned striatum were significantly decreased (P<0.001) in comparison with those in the controls or in the unlesioned side 3 weeks after lesioning while the extracellular level of MLT in the lesioned striatum in these corresponding time intervals distinctly increased when compared with those in the controls (P<0.05). The tissue MLT contents increased in the bilateral striata in different degrees at 6 weeks post-lesion (P<0.05). Moreover, increased MLT levels correlate well with rotations or DA changes in the lesioned striatum., Significance: These data suggest that 6-OHDA lesion manipulates the MLT secretion pattern. Increased striatal MLT level by a unilateral intracerebral injection of 6-OHDA may play dual roles in the progression of PD in rats., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Reproductive experience (RE), i.e. pregnancy and lactation, induces physiological changes in mammals. Recent data show that neuroimmune interactions are modulated by a diversity of events involving neurotransmitters and neuropeptides. These molecules, particularly dopamine (DA), were reported to mediate the relevant cross talk between immune and neuroendocrine systems. Moreover, DA-mediated regulation of leukocyte function is a reasonable approach to investigate the DA-operated regulatory switch for immune-competent cells, such as macrophages. Therefore, the goals of the present study were to determine the effects of RE on: (1) dopaminergic function through hypothalamic levels of DA, dihydroxyphenylacetic acid (DOPAC), homovanilic acid (HVA), serotonin (5-HT), and 5-hydroxyindole acetic acid (5-HIAA); (2) basal levels of circulating prolactin (PRL); and (3) activity of peritoneal macrophage (phagocytosis and oxidative burst). A total of 16 adult (200-250 g) female Wistar rats were used, divided in two groups: nulliparous and primiparous. Approximately 2-3 weeks after weaning pups from the primiparous group, both groups of rats were tested. The findings indicate that: (1) DOPAC concentrations, DOPAC/DA and HVA+DOPAC/DA ratios decreased in primiparous rats as compared to virgin rats, (2) primiparous rats showed significantly lower serum PRL levels, and (3) phorbol miristate acetate (PMA)-induced oxidative burst was decreased in peritoneal macrophage from primiparous rats as compared to virgin rats. To test the possible positive correlation between serum levels of PRL and the intensity of oxidative burst by peritoneal macrophage, an extra experiment was done with adult virgin female rats treated with domperidone, an antagonist of DA receptors. Domperidone-treated animals showed increased serum levels of PRL and simultaneous increase in peritoneal macrophage oxidative burst. Thus, suggesting an indirect participation of hyperprolactinemia, induced by this treatment in peritoneal macrophage activity of female rats. These results suggest that a previous RE can modulate the activity of dopaminergic hypothalamic systems, while decreasing PRL serum levels and the oxidative burst of peritoneal macrophage. The neurochemical and hormonal RE-induced changes correlate with the immune alterations.

To date, UCM707, (5Z,8Z,11Z,14Z)-N-(3-furylmethyl)eicosa-5,8,11,14-tetraenamide, has the highest potency and selectivity in vitro and in vivo as inhibitor of the endocannabinoid uptake. Its biochemical, pharmacological and therapeutic properties have been intensely studied recently, but the information on its capability to modify neurotransmitter activity, which obviously underlies the above properties, is still limited. In the present study, we conducted a time-course experiment in rats aimed at examining the neurochemical effects of UCM707 in several brain regions following a subchronic administration (5 injections during 2.5 days) of this inhibitor in a dose of 5 mg/kg weight. In the hypothalamus, the administration of UCM707 did not modify GABA contents but reduced norepinephrine levels at 5 h after administration, followed by an increase at 12 h. Similar trends were observed for dopamine, whereas serotonin content remained elevated at 1 and, in particular, 5 and 12 h after administration. In the case of the basal ganglia, UCM707 reduced GABA content in the substantia nigra but only at longer (5 or 12 h) times after administration. There were no changes in serotonin content, but a marked reduction in its metabolite 5HIAA was recorded in the substantia nigra. The same pattern was found for dopamine, contents of which were not altered by UCM707 in the caudate-putamen, but its major metabolite DOPAC exhibited a marked decrease at 5 h. In the cerebellum, UCM707 reduced GABA, serotonin and norepinephrine content, but only the reduction found for norepinephrine at 5 h reached statistical significance. The administration of UCM707 did not modify the contents of these neurotransmitters in the hippocampus and the frontal cortex. Lastly, in the case of limbic structures, the administration of UCM707 markedly reduced dopamine content in the nucleus accumbens at 5 h, whereas GABA content remained unchanged in this structure and also in the ventral-tegmental area and the amygdala. By contrast, norepinephrine and serotonin content increased at 5 h in the nucleus accumbens, but not in the other two limbic structures. In summary, UCM707 administered subchronically modified the contents of serotonin, GABA, dopamine and/or norepinephrine with a pattern strongly different in each brain region. So, changes in GABA transmission (decrease) were restricted to the substantia nigra, but did not appear in other regions, whereas dopamine transmission was also altered in the caudate-putamen and the nucleus accumbens. By contrast, norepinephrine and serotonin were altered by UCM707 in the hypothalamus, cerebellum (only norepinephrine), and nucleus accumbens, exhibiting biphasic effects in some cases.

In a previous study, we demonstrated that iptakalim (Ipt) significantly ameliorated hypolocomotion and catalepsy induced by haloperidol and rotenone in rats. In order to further understand the mechanism(s), using a rat model of Parkinson's disease (PD) established by unilateral 6-hydroxydopamine (6-OHDA) administration to the substantia nigra pars compacta (SNpc) and reverse microdialysis techniques with high performance liquid chromatography (HPLC), we investigated the effects of Ipt on extracellular levels of glutamate, dopamine (DA) and its metabolite dihydroxyphenylacetic acid (DOPAC) in the striatum of conscious and freely moving rats. The results indicated that unilateral 6-OHDA-lesioned rats have a significantly higher level of extracellular glutamate and a lower level of extracellular DOPAC in the lesioned-side of the striatum, and a lower level of extracellular DA in both sides of the striatum compared to the striatum of control rats. Ipt reduced extracellular glutamate levels in both sides of striatum of the lesioned and control rats in a concentration-dependent manner. Ipt, at lower concentrations (0.01, 0.1, 1 microM), enhanced extracellular DA levels in the lesioned-side striatum of the unilateral 6-OHDA-lesioned rats, while causing no significant changes in the intact side striatum, and even a significant decline in striatum of control rats at higher concentrations of Ipt (10, 100 microM). In addition, Ipt also caused a significant decline in the extracellular DOPAC levels in the lesioned-side striatum of unilateral 6-OHDA-lesioned rats. These data suggest that the major mechanism underlying the ameliorative effects of Ipt on the behavior in 6-OHDA-lesioned rats is the alteration of levels of extracellular neurotransmitters, such as glutamate and DA in the striatum of unilateral 6-OHDA-lesioned rats.

The present work showed that the intrastriatal injection of 6-OHDA significantly decreases DA, DOPAC and HVA levels in that rat brain structure. Although there is also a decrease in 5-HT levels no changes were observed in 5-HIAA levels as compared to controls. On the other hand, melatonin (2, 5, 10 and 25 mg/kg. i.p., daily for 7 days) treatment starting 1 h after 6-OHDA lesions, partially reverses the decreases caused by 6-OHDA lesions on these neurotransmitter levels, and contents were brought to approximately 50% of that observed in the contralateral sides of controls or of melatonin treated group. Melatonin was more efficient at the doses of 5 and 10 mg/kg, i.p., and effects were similar between the lowest and highest doses characteristic of a bell-shaped type of response. The apomorphine-induced rotational behavior (3 mg/kg, i.p.) was blocked by 60, 89, 78 and 47% after the doses of 2, 5, 10 and 25 mg/kg, i.p., respectively. Similarly, in this case the doses of 5 and 10 mg/kg were also more efficient. Melatonin (5 mg/kg) produced an upregulation of D1 receptors associated with a decrease in Kd value. While no change was observed in maximum density of D2 receptors, the Kd value was also decreased.

Muscarinic acetylcholine receptors (M1-M5) play important roles in the modulation of many key functions of the central and peripheral nervous system. To explore the physiological roles of the two Gi-coupled muscarinic receptors, we disrupted the M2 and M4 receptor genes in mice by using a gene targeting strategy. Pharmacological and behavioral analysis of the resulting mutant mice showed that the M2 receptor subtype is critically involved in mediating three of the most striking central muscarinic effects, tremor, hypothermia, and analgesia. These studies also indicated that M4 receptors are not critically involved in these central muscarinic responses. However, M4 receptor-deficient mice showed an increase in basal locomotor activity and greatly enhanced locomotor responses following drug-induced activation of D1 dopamine receptors. This observation is consistent with the concept that M4 receptors exert inhibitory control over D1 receptor-mediated locomotor stimulation, probably at the level of striatal projection neurons where the two receptors are known to be coexpressed. These findings emphasize the usefulness of gene targeting approaches to shed light on the physiological and pathophysiological roles of the individual muscarinic receptor subtypes.

Central administration of gamma-aminobutyric acid (GABA) has been shown to stimulate the secretion of prolactin (PRL). Whether GABA acts via dopamine, the major PRL-inhibiting hormone, and which GABA receptor type(s) is involved have not been ascertained. Both GABA(A) and GABA(B) receptor agonists and/or antagonists were administered centrally in this study and their effects on both basal and diurnal changes of tuberoinfundibular dopaminergic (TIDA) neuronal activity were determined by measuring the concentration of 3,4-dihydroxyphenylacetic acid (DOPAC) in the median eminence (ME). Serum PRL level was determined by RIA. Ovariectomized, estrogen-primed Sprague-Dawley rats implanted with intracerebroventricular (icv) cannulae were used. Muscimol (1 ng/3 microl/rat, icv), a GABA(A) receptor agonist, but not baclofen (1-100 ng/3 microl/rat, icv), a GABA(B) receptor agonist, injected in the morning significantly lowered and elevated ME DOPAC and serum PRL levels, respectively at 15 and 30 min. Lower and higher doses of muscimol were not effective. The effects of muscimol could also be prevented by co-administration of bicuculline (0.1-10 ng/3 microl, icv), a GABA(A) receptor antagonist. When bicuculline (10-500 ng/3 microl, icv) was given in the afternoon (at 1500 h), it significantly reversed the lowered ME DOPAC level in the afternoon and prevented the concurrent PRL surge. We conclude that endogenous GABA acting through GABA(A) receptors may play a significant role in the control of basal and diurnal changes of TIDA neuronal activity, and in turn, PRL secretion.

Previous report from our laboratory showed that morphine produces a stimulatory effect of hypothalamic noradrenaline (NA) turnover concurrently with enhanced pituitary-adrenal response after its acute injection and during withdrawal. In the present work we have studied the effects of acute and chronic administration of the kappa agonist U-50,488H as well as the influence of U-50,488H withdrawal on the activity of hypothalamic NA and dopamine (DA) neurons and on the activity of hypothalamic-pituitary-adrenal (HPA) axis. A single dose of U-50,488H (15 mg/kg i.p.) significantly increased hypothalamic NA and decreased DA turnover at the time of an enhanced corticosterone release. Rats rendered tolerant to the kappa agonist by administration of U-50,488H twice a day for 4 days showed no changes in corticosterone secretion. Additionally, a decrease in both hypothalamic MHPG (the cerebral NA metabolite) production and NA turnover was observed, whereas DOPAC concentration and DA turnover were enhanced, which indicate the development of tolerance towards the neuronal and endocrine actions of U-50,488H. After naloxone (3 mg/kg s.c.) administration to U-50,488H-tolerant rats, we found neither behavioural signs of physical dependence nor changes in hypothalamic catecholaminergic neurotransmission. In addition, corticosterone secretion was not altered in U-50,488H withdrawn rats. Present data clearly indicate that tolerance develops towards the NA turnover accelerating and DA turnover decreasing effect of U-50,488H. Importantly and by contrast to mu agonists, present results demonstrate that U-50,488H withdrawal produce no changes in hypothalamic catecholamines turnover or in corticosterone release (an index of the hypothalamus-pituitary-adrenal activity), which indicate the absence of neuroendocrine dependence on the kappa agonist. As has been proposed, this would suggest that the mu and the kappa receptor be regulated through different cellular mechanisms, as kappa agonists have a lower proclivity to induce dependence.

A selective inhibitor of the carrier-mediated transport of endogenous cannabinoids, N-(4-hydroxyphenyl)-arachidonylethanolamide (AM404), has been recently synthesized and proposed as a useful tool for studying the physiological effects of endogenous cannabinoids and as a potential therapeutic agent in a variety of diseases. In the present study, we have examined the effects of this compound in two important brain processes in which a role for anandamide and other endogenous cannabinoids has been claimed: neuroendocrine regulation and extrapyramidal motor activity. A single and well-characterized dose of AM404, which presumably resulted in a significant elevation of the levels of endogenous cannabinoids, produced a marked decrease in plasma prolactin (PRL) levels, with no changes in luteinizing hormone (LH) levels. This decrease in PRL levels was accompanied by an increase in the activity of tyrosine hydroxylase (TH) in the medial basal hypothalamus. Both decreased PRL secretion and increased hypothalamic TH activity have been reported to occur after the administration of anandamide. Administration of AM404 also produced a marked motor inhibition in the open-field test, as also reported for anandamide, with a decrease in ambulatory and exploratory activities and an increase in the time spent in inactivity. This was accompanied by a decrease in the activity of TH in the substantia nigra, an effect also previously observed for anandamide.

Using in vivo microdialysis and flow-injection, we evaluated the production of nitric oxide (NO) in the striatum and substantia nigra of freely moving rats in response to challenge doses of L-dihydroxyphenylalanine (L-DOPA) by measuring the NO metabolite levels of nitrate and nitrite. The dopamine and 3,4-dihydroxyphenylacetic acid (DOPAC) concentrations also were determined in the same perfusates. Intraperitoneal injection of L-DOPA produced a significant, dose-dependent increase in the extracellular levels of dopamine and DOPAC in these areas, but did not modify the extracellular levels of the NO metabolites. An acute dose of L-DOPA does not appear to facilitate NO production in the rat striatum and substantia nigra.

The present study reports on the presence of type A and B monoamine oxidase (MAO) activity and their sensitivity to selective MAO-A and MAO-B inhibition by Ro 41-1049 and lazabemide, respectively, in homogenates of isolated rat renal tubules. Non-linear analysis of the saturation curve of H-5-hydroxytryptamine (3H-5-HT ) deamination revealed a Km of 351+/-71 microM (n=4) and a Vmax of 25+/-2 nmol mg protein(-1) h(-1). Deamination of 14C-beta-phenylethylamine (14C-beta-PEA) was also a saturable process yielding Km values of 58+/-12 microM and Vmax values of 24+/-2 nmol mg protein(-1) h(-1). Ro 41-1049 produced a concentration-dependent inhibition of 3H-5-HT deamination with a Ki of 24 nM. Deamination of 14C-beta-PEA was found to be reduced by lazabemide in a concentration-dependent manner with a Ki value of 17 nM. The effect of these selective MAO inhibitors on dopamine fate and DOPAC formation in isolated tubular epithelial cells was also studied. In these studies a clear inhibition of DOPAC formation was observed with Ro 41-1049 (250 nM), while 250 nM lazabemide was found not to increase the accumulation of newly-formed DA in those tubular epithelial cells loaded with 50 microM L-DOPA. In conclusion, the results presented here confirm the presence of both MAO-A and MAO-B activity in renal tubular epithelial cells, that MAO-A is the predominant enzyme involved in the deamination of the natriuretic hormone dopamine and that the deamination of newly-formed dopamine is a time-dependent process which occurs early after the decarboxylation of L-DOPA.

Studies performed in our laboratory indicate that the adrenal deprivation during gestation can greatly influence the fetal catecholamines development in several cerebral areas. The present study was undertaken to determine whether the administration of metyrapone to pregnant rats affects the content of monoamines in fetal brain at term. To test wether the content of monoamines in fetal brain is regulated, at least in part, by endogenous glucocorticoids, pregnant rats were injected for 5 days prior to delivery with metyrapone, an adrenal 11-beta-steroid hidroxylase inhibitor which crosses the placenta and blocks endogenous glucocorticoid synthesis, or saline. On day 21 of gestation, delivery of all animals was accomplished by cesarean section. The encephalons were extracted and immediately dissected in metencephalon, mesencephalon, diencephalon and telencephalon. Monoamine determination was carried out using HPLC-ED. The results obtained indicate that the metyrapone treatment increases both DA and 5-HT and their metabolites in the brain studied areas.

The effects of Selenium (Se) on central dopaminergic function were examined in male Sprague-Dawley rats. In this experiment, animals were implanted with microdialysis probes and dialysates were analyzed for dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA). After reaching baseline values, sodium selenite was either injected intraperitoneally (i.p.) or directly infused into the striatum (ST) or nucleus accumbens (NA). Se administration of 3.0 mg/kg (i.p.) significantly increased (70%) DA overflow in the ST. Meanwhile direct Se perfusion (10 mM) also caused a significant elevation of synaptic DA concentrations in the ST and NA. Levels of DOPAC and HVA were minimally affected in all studies. In order to test for the effects of DA receptor activation, animals were pretreated with quinpirole (0.5 mg/kg, s.c.), an hour prior to Se (10 mM) infusion through the probe. It was found that quinpirole pre-treatment reduced Se-induced changes in DA concentrations. It was concluded from the present study that Se's central action might be related to its ability to potentiate DA function.

We investigated whether enhanced spontaneous locomotor activity in streptozotocin (STZ)-induced diabetic mice is associated with serum glucose levels. Furthermore, the role of factor(s) derived from spleen cells on the enhanced spontaneous locomotor activity in diabetic mice was also examined. Serum glucose levels were significantly increased on day 4 after STZ administration and remained increased at 7, 14, 28 and 56 days after STZ administration. A significant increase in spontaneous locomotor activity in diabetic mice as compared to that in non-diabetic mice was observed 4, 7, 14 or 28 days after STZ treatment. The increase in spontaneous locomotor activity in diabetic mice was not observed when insulin was chronically administered to mice 3 to 7 days after administration of STZ. Splenectomized diabetic mice, which were operated upon 7 days after STZ administration, still had a significantly higher spontaneous locomotor activity than non-diabetic mice. In contrast, spontaneous locomotor activity in diabetic mice that were splenectomized either before or 3 days after administration of STZ returned to the levels in non-diabetic mice. The rate of dopamine (DA) turnover in the limbic forebrain of diabetic mice on day 14 was significantly higher than that in age-matched non-diabetic. Although the rate of DA turnover in the diabetic mice that were splenectomized 7 days after STZ administration was significantly higher than that in non-diabetic mice, an increased the rate of DA turnover was not observed when mice were splenectomized either before or 3 days after administration of STZ. These results suggest that the lasting hyperglycemia may be responsible for their hyperlocomotion in diabetic mice within 14 days sfter STZ treatment, perhaps due to an increase in dopamine release in mesolimbic dopamine systems. Furthermore, factor(s) derived from spleen cells of diabetic mice during the incipient stages of diabetes may play a role in the enhancement of dopaminergic neurotransmission.

Lamotrigine (LTG), an anticonvulsive drug, was tested for its effects on striatal content of DA and its metabolites, DOPAC and HVA, in audiogenic seizure-resistant (ER) and audiogenic seizure-prone (EP) lines of Balb/c mice. A single dose of LTG (20 mg/kg) prevented audiogenic seizures in seizure-prone mice, while reducing substantially the striatal content of the DA metabolite, DOPAC (to less than 50% of saline-injected controls) in both seizure-resistant and seizure-prone mice. LTG administration also resulted in significant reduction of striatal content of HVA. The in situ activity of tyrosine hydroxylase (TH) in extracts of striatum was significantly reduced by LTG administration in both ER and EP mice. These data show that DA synthesis in the striatum of mice is substantially reduced by LTG administration.

The effects of central administration of two serotonin receptor agonists, 8-OH-DPAT and mCPP, on tuberoinfundibular dopaminergic (TIDA) neuron activity and serum prolactin (PRL) levels in ovariectomized, estrogen-treated rats were determined. 8-OH-DPAT dose-dependently (0.1-10 microgram/rat, icv) stimulated serum PRL levels, and depressed serotonergic neuron activity in 30 min. However, the TIDA neuron activity was not affected at all. Similar treatment of mCPP was less effective than 8-OH-DPAT: only the highest dose of mCPP (10 microgram) stimulated PRL secretion and inhibited serotonergic neuron activity. No change in TIDA neuron activity was observed either. We conclude that central serotonin acts on 5-HT1 receptors to stimulate the PRL secretion, which may not involve the TIDA neurons.

In the present study, we compared the changes in dopamine (DA) release and clearance in the striatum after unilateral 6-hydroxydopamine (6- OHDA) lesioning of the nigrostriatal DA pathway in urethane-anesthetized rats. High-speed in vivo chronoamperometric recording techniques using Nafion-coated carbon fiber electrodes were used to evaluate extracellular DA concentration. We found that DA release, induced by local KCl application in the striatum, was maximally suppressed 21 days after lesioning. DA clearance was also affected; however, the maximal effect occurred much earlier than changes in DA release. We found that 3-7 days after lesioning, extracellular DA concentration was significantly higher in the lesioned striatum after locally applying DA. Local application of nomifensine, which blocks high affinity DA uptake, did not further potentiate the amplitude and duration of DA overflow in these animals, suggesting that the high affinity uptake of DA was abolished 3-7 days after 6-OHDA lesioning. In conclusion, our data suggest that the time courses of changes in clearance and the release of DA are differentially affected by this selective neurotoxin.

We examined the effect of 7-OH-DPAT on spontaneous locomotor activity that is enhanced in diabetic mice. 7-OH-DPAT (0.1-30 mg/kg, s.c.) dose-dependently decreased the enhanced spontaneous locomotor activity in diabetic mice, but not in nondiabetic mice. When mice were pretreated with of 7-OH-DPAT (0.1 mg/kg), morphine-induced enhanced locomotor activity in nondiabetic mice, but not in diabetic mice, was significantly reduced. Furthermore, 7-OH-DPAT (0.1 mg/kg) significantly attenuated the morphine-induced increase in DA turnover in both nondiabetic and diabetic mice. However, DA turnover was significantly greater in diabetic mice than in nondiabetic mice. Thus, it is likely that the enhanced spontaneous locomotor activity in diabetic mice may be partially related to the down-regulation of D3-receptor-mediated modulation of dopamine release in the limbic area.

During 30 minutes of global forebrain ischemia in rats, striatal intracellular levels of dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) increase at the same time extracellular levels, measured by microdialysis, decrease. If DOPAC and HVA were free to diffuse out of cells, these changes in opposite directions should not occur. These data support the theory that a membrane potential-dependent active transport system is required for DOPAC and HVA to leave striatal cells. During ischemia and subsequent to ATP depletion, neurons and glia depolarize and active transport systems fail. DOPAC and HVA, which are still being produced inside neurons and possibly glia, can not be transported out of these cells and accumulate intracellularly.

The dopamine (DA) uptake inhibitor, GBR 12909 (GBR) and a neonatal dopaminergic denervated rat model were used as tools to study the influence of DA on the serotonin (5HT) system in the striatum. The striatal levels of the amines and their acid metabolites (dihydroxyphenylacetic acid, DOPAC; 5-hydroxyindoleacetic acid, 5HIAA) were determined by HPLC. The administration of a single dose (20 mg/kg) of GBR failed to affect the steady-state levels of the amines or metabolites. Repeated administration of GBR (20 mg/kg/day) for 2 or 4 days decreased DA and DOPAC; only the 4-day regimen decreased 5HT and increased 5HIAA levels. The neonatal dopaminergic lesion with 6-hydroxydopamine (6OHDA) depleted (> 95%) DA and DOPAC and increased 5HT and 5HIAA levels in the striatum. Administration of GBR (20 mg/kg/day for 4 days) to lesioned animals failed to influence the lesion-induced increases in 5HT and 5HIAA levels. The results suggest GBR decreases the steady-state levels of DA, resulting in a compensatory increase in the turnover of 5HT that is dependent on the presence of intact dopaminergic terminals. Thus, the effect of GBR on 5HT turnover is indirect. The studies provide further support for a prominent dopaminergic influence on striatal 5HT metabolism.

Rats of the Lewis (LEW) strain show a greater preference for drugs of abuse than do Fischer 344 (F344) rats. The in vivo microdialysis procedure was used to examine basal and cocaine-evoked extracellular (EC) levels of dopamine (DA), DOPAC, and HVA in the nucleus accumbens (NAc) of F344 and LEW rats. The basal EC levels of the DA metabolites DOPAC and HVA in the NAc were markedly lower in LEW than in F344 rats. Although the increase in EC DA after 3, 10, or 30 mg/kg, i.p. of cocaine was similar in both strains, LEW rats had a smaller peak DA elevation followed by a slower return to basal DA levels at the 30 mg/kg dose. The neurochemical differences observed may contribute to the strain differences in the behavioral response to cocaine.

The brain regional dopamine (DA) and serotonin (5-HT) levels and turnover were studied in the spontaneously hypertensive rat (SHR), following voluntary, long-lasting (7 weeks) wheel-running exercise. Groups of rats were sacrificed 1-2 h, 23-24 h or 47-48 h after termination of the last running session, and the cerebral tissue levels of 5-HT, 5-HIAA, DA and DOPAC were determined and compared to corresponding levels obtained in sedentary controls. In the exercised animals, there was a selective decrease in the limbic forebrain levels of DOPAC in the immediate post-exercise period (1-2 h), while the DA turnover (DOPAC/DA ratio) was not altered. In addition, the 5-HT and 5-HIAA levels in the serotoninergic nerve terminal limbic forebrain and the 5-HT turnover (5-HIAA/5-HT ratio) in the cell body-containing brain stem areas were decreased in the immediate post-exercise period. No significant changes in the DA, DOPAC, 5-HT or 5-HIAA levels, nor in the DA and 5-HT turnover, were obtained in the remainder of the brain regions analyzed, regardless of time after termination of running. Taken together, the present study indicates that voluntary exercise in the SHR gives rise to differentiated regional effects on brain DA and 5-HT levels and turnover, thus supporting the view that changes in central monoaminergic activity are involved in the functional effects of long-term exercise. Interestingly, the psychomotor-associated limbic forebrain areas appeared particularly susceptible.

Dopamine (DA) metabolites in microdialysates from the medial preoptic area (MPOA) of male rats increased during copulation. These increases were not observed during eating of a highly palatable food, or if the animal failed to copulate, or if the microdialysis probe was anterior or dorsal to the MPOA. The only two animals with measurable serotonin (5-HT) levels while the female was present were also the only two that either failed to copulate or copulated but failed to ejaculate. These data are consistent with previous evidence for a facilitative role of MPOA DA in the control of male sexual behavior; however, 5-HT activity in the MPOA may impair copulation.

The present study examined the effect of prolonged morphine treatment on striatal dopamine (DA) release and metabolism, during the initial phase of the development of morphine dependence. Sprague-Dawley rats were implanted with chronic guides for microdialysis of the striatum. Morphine (two 75-mg pellets, subcutaneous implant) or placebo was given (12 hr) to pentobarbital anesthetized animals. Following recovery from anesthesia, morphine physical dependence was verified by the naloxone-evoked abstinence syndrome. Morphine produced significant increases in the dialysate level of DA nad its metabolites (DOPAC and HVA) above baseline compared to placebo treatment. HVA levels began to increase immediately following morphine administration, whereas DA and DOPAC levels began to increase after a latency of one and three hr, respectively. Morphine effects on striatal DA metabolism included changes in the metabolic disposition of DA. Increases in HVA concentration accompanied increases in DOPAC concentration up to a threshold value of DOPAC efflux; further increases in DOPAC level were associated with decreases in HVA level. These in vivo data suggest that morphine-induced changes in the regulation of striatal dopaminergic function may be an important component of the development of physical dependence.

The effects of neurotensin on the activity of hypothalamic tuberoinfundibular and periventricular-hypophysial dopaminergic (DA) neurons, and on the secretion of pituitary hormones that are tonically regulated by these neurons (i.e. prolactin and alpha-melanocyte-stimulating hormone [alpha MSH], respectively) were examined in estrogen-primed ovariectomized rats. The activity of tuberoinfundibular and periventricular-hypophysial DA neurons was estimated by measuring concentrations of the dopamine metabolite 3,4-dihydroxyphenylacetic acid (DOPAC) in the terminals of these neurons in the median eminence and intermediate lobe of the posterior pituitary, respectively. Intracerebroventricular administration of neurotensin caused a dose- and time-related increase in DOPAC concentrations in both the median eminence and intermediate lobe, and a concurrent decrease in plasma levels of prolactin and alpha MSH. These results suggest that neurotensin-induced inhibition of secretion of prolactin and alpha MSH from the pituitary may be due to the stimulatory action of this neuropeptide on the release of dopamine from tuberoinfundibular and periventricular-hypophysial neurons.

The effects of histamine on prolactin secretion and the activity of tuberoinfundibular dopaminergic (DA) neurons were examined in male rats. Tuberoinfundibular DA neuronal activity was estimated in situ by measuring the metabolism [concentration of 3,4-dihydroxyphenylacetic acid (DOPAC)] and synthesis [accumulation of 3,4-dihydroxyphenylalanine (DOPA) after administration of a decarboxylase inhibitor] of dopamine in the median eminence. Intracerebroventricular (icv) injection of histamine produced a dose- and time-dependent increase in plasma prolactin levels but had no effect on DOPA accumulation or DOPAC concentrations in the median eminence. These results indicate that the stimulation of prolactin secretion following icv histamine is not mediated by an inhibition of tuberoinfundibular DA neurons.

Pretreatment of rats with the noradrenergic neurotoxin DSP-4 selectively reduced regional levels of noradrenaline in the brain by more than 75%, and decreased the concentration of endogenous DA in microdialysates of the caudate nucleus and nucleus accumbens by 52% and 28%, respectively. Results support the hypothesis that central noradrenergic mechanisms facilitate nigrostriatal and mesolimbic dopamine transmission in vivo.

The administration of amphetamine to rats pretreated with iprindole to inhibit the metabolism of amphetamine results in a long-lasting depletion of striatal dopamine and its metabolites, DOPAC and HVA. Pretreatment with MK801, a noncompetitive antagonist of the NMDA (N-methyl-D-aspartate) subclass of excitatory amino acid receptors, antagonized the depletion of striatal dopamine, DOPAC and HVA 3 days after a single dose of amphetamine in iprindole-treated rats. MK801 pretreatment was effective up to 4 hours but not at 8 or 24 hours in preventing amphetamine effects on striatal dopamine, DOPAC and HVA.

The effect of ibogaine hydrochloride on locomotor stimulation induced by d-amphetamine sulfate was tested in male C57BL/6By mice and in female Sprague-Dawley rats. In mice, locomotor stimulation induced by d-amphetamine at 1 or 5 mg/kg s.c. was reduced by prior administration of one or two injections of ibogaine (40 mg/kg), given 2 or 18 hours earlier. This reduction in locomotor activity persisted for two days. Locomotor stimulation induced by a higher dose (10 mg/kg) of d-amphetamine was not reduced by such prior administration of ibogaine. A lower dose of ibogaine (20 mg/kg) did not reduce the subsequent locomotor activity induced by d-amphetamine. Ibogaine decreased striatal dopamine levels, while d-amphetamine increased them. Ibogaine treatment (2 x 40 mg/kg, 18 hours apart) induced a decrease by 30% in the level of striatal dopamine and its metabolites measured in tissue extracts 3 hours after the second ibogaine injection. One hour after d-amphetamine (5 mg/kg) administration, the level of striatal dopamine increased by 26%. Although the level of striatal dopamine was initially lower in the ibogaine-pretreated mice, d-amphetamine (5 mg/kg) administration induced an increase in striatal dopamine and its metabolites. The effect of ibogaine seems to be species specific, since in rats pretreated with ibogaine 18 hours before d-amphetamine, locomotor stimulation induced by d-amphetamine was further increased. In addition, the in vitro electrical-evoked release of [3H]dopamine from striatal tissue was either unchanged or inhibited in the presence of d-amphetamine, and after ibogaine pretreatment in vivo, the release of tritium in the presence of d-amphetamine was inhibited or stimulated in mice and rats, respectively.

The present study has examined the effects of systemically administered MPTP and MPP+ upon striatal DA and Dopac of C57 mice, also treated concurrently with either saline or reserpine. MPTP followed by saline did not affect DA level but decreased that of Dopac only at 5.0 mg/kg and higher dosages. The potency of MPTP affecting DA increased greatly when the neurotoxicant was followed by either 5.0 or 10.0 mg/kg reserpine; MPTP at 0.10 mg/kg and higher dosages significantly reversed the DA depleting effects of reserpine. MPP+ (1.0 or 10.0 mg/kg) with saline did not affect either DA or Dopac. In contrast, MPP+ at 0.10 mg/kg and higher dosages, when followed by 10.0 mg/kg reserpine, dose-dependently enhanced the DA depleting effects of reserpine. In agreement with the earlier results obtained in vitro, the present study indicates that MPTP administration at trace level dosages may lead to an inhibition of MAO in vivo. The effect of systemically given MPP+ on DA, however, appears to be more complex in nature, conceivably comprised of actions at the striatal neurones including the intraneuronal vesicles and, possibly, at the substantia nigra which may affect striatum in turn. That MPP+ may have reached brain areas in these experiments is also indicated by the observation of a significant striatal level of 3H-MPP+ after its systemic administration. In conclusion, irrespective of MPTP and MPP+ action mechanisms, trace levels of these neurotoxicants appear to affect brain dopamine neurons.

The activities of periventricular-hypophysial dopaminergic (DA) neurons were compared in male and female rats by measuring dopamine synthesis (accumulation of 3,4-dihydroxyphenylalanine [DOPA] after inhibition of L-aromatic amino acid decarboxylase) and metabolism (concentrations of 3,4-dihydroxyphenylacetic acid [DOPAC]) in terminals of these neurons in the intermediate lobe of the pituitary. For comparison, the synthesis and metabolism of dopamine in the neural lobe of the pituitary and median eminence were also determined. The concentrations of DOPAC and accumulation of DOPA were higher in females than in males in both the intermediate lobe and median eminence, revealing a sexual difference in the basal activity of periventricular-hypophysial and tuberoinfundibular DA neurons. In contrast, there were no differences between male and female rats in activity of DA neurons terminating in the neural lobe. One week following gonadectomy, DOPA accumulation in the median eminence was decreased in females and increased in males, but remained unchanged in the intermediate lobe. These results indicate that sexual differences in the activity of periventricular-hypophysial DA neurons terminating in the intermediate lobe are not dependent upon the presence of circulating gonadal steroids, and in this respect, these neurons differ from tuberoinfundibular DA neurons.

Push-pull perfusion technique was used to infuse IL-1 beta into and collect perfusate from the medial basal hypothalamus of freely moving male rats. Dopamine (DA) and its metabolite, dihydroxyphenylacetic acid (DOPAC), were measured in the perfusate using high performance liquid chromatography with electrochemical detection. In the control group, release rates of DA and DOPAC decreased and were 62% and 40%, respectively, below pretreatment levels after 325 min. In contrast, treatment with 50 ng of IL-1 beta produced substantial reductions in these decreases, and treatment with 100 ng of IL-1 beta produced increases of up to 118% and 89% in the release rates of DA and DOPAC, respectively. It is concluded that IL-1 beta affects the metabolism of catecholamines (and probably other neurotransmitters) in the brain, which, in turn, mediate its central and neuroendocrine actions.

We kinetically characterized D2 receptors in thalami pooled from a group of Sprague-Dawley rats and then determined thalamic levels of dopamine (DA), homovanillic acid (HVA), dihydroxyphenylacetic acid (DOPAC), and norepinephrine (NE) in relation to a measure of thalamic DA D2 receptor densities in another group of rats. The equilibrium dissociation constant (kd) was estimated as 0.1 nM by three independent methods, while the Bmax for thalamic D2 receptors was found to be 6.4 fmol/mg p using 3H-spiperone as ligand and ketanserin to occlude 5HT2 binding. Kinetic constants were in agreement with previously reported kinetic data from rodent caudate-putamen. This suggests that thalamic D2 receptors are similar to D2 receptors from other brain areas. Mean thalamic levels of DA (22.6 ng/mg p), DOPAC (1.19 ng/mg p) and HVA (0.31 ng/mg p) concur with previous reports of a sparse distribution of thalamic DA neurons. D2 receptor densities were positively correlated with DA metabolites DOPAC (P less than .05; r = 0.423) and HVA (P less than .05; r = 0.368), but not DA or NE. These results establish fundamental characteristics of thalamic DA neurotransmission to assist in the investigation of behavioral pharmacology of this area.

The effect of the kappa opioid receptor antagonist nor-binaltorphimine (NOR-BNI) was examined on the activity of dopamine (DA) neurons comprising the nigrostriatal, mesolimbic, and tuberohypophysial systems in the male rat. DA neuronal activity was estimated by measuring: (1) the concentration of the DA metabolite 3,4-dihydroxyphenylacetic acid and, (2) the accumulation of 3,4-dihydroxyphenylalanine after administration of a decarboxylase inhibitor in brain (striatum, nucleus accumbens) and pituitary regions (intermediate lobe, neural lobe) containing terminals of these neurons. The intracerebroventricular administration of NOR-BNI produced a dose- and time-related increase in the activity of tuberohypophysial DA neurons, but failed to alter the activity of nigrostriatal or mesolimbic DA neurons. The ability of NOR-BNI to enhance the activity of tuberohypophysial DA neurons was blocked by the kappa opioid agonist U-50,488. These results indicate that NOR-BNI, acting on kappa opioid receptors, activates tuberohypophysial DA neurons projecting to the neural and intermediate lobes of the pituitary.

Although it had been suggested that prolactin (PRL) modulates the dopaminergic receptor sensitivity in extrahypothalamic areas, recent studies have questioned this role. We studied the effects of PRL on the receptor sensitivity in the striatum and the limbic forebrain, analyzing the number of D1 and D2 receptors and the amount of their second messenger, cyclic-adenosine monophosphate (cAMP). Tyrosine hydroxylase (TH) activity and dopamine (DA) and L-3,4-dihydroxyphenylacetic acid (DOPAC) content were also measured as indices of presynaptic activity. The study was carried out in male rats submitted to either acute (PRL injection) or chronic (pituitary grafts or diethylstilbestrol (DES)-induced pituitary tumors) rises of plasma PRL levels. The results showed a common lack of effect of PRL on the dopaminergic receptor sensitivity in both brain areas and, only some few effects on presynaptic activity in the striatum. Thus, grafted rats showed a slight decrease in DA content in the striatum, but neither D1 and D2 receptor number and cAMP content nor DOPAC content and TH activity, were modified, whereas DES animals exhibited no changes in all the parameters studied. A single injection of ovine PRL caused a decrease in DOPAC content and an increase in TH activity in the striatum. In the case of the limbic area, both chronic and acute hyperprolactinemia failed to alter any of the indices studied. In summary, we cannot support the view that PRL plays a role as modulator of dopaminergic receptor sensitivity. The only effects were always produced at the presynaptic level on the striatum, and after acute treatment, which supports the possible development of tolerance after chronic changes in peripheral PRL levels.

A mathematical framework was developed to provide a quantitative basis for either in vivo tissue or in vitro microdialysis. Established physiological and mass transport principles were employed to obtain explicit expressions relating dialysate concentration to tissue extracellular concentration for in vivo applications or external medium concentrations for in vitro probe characterization. Some of the important generalizations derived from the modeling framework are: (i) the microdialysis probe can perturb the spatial concentration profile of the substance of interest for a considerable distance from the probe, (ii) for low molecular weight species the tissue is generally more important than the probe membrane in determining the dialysate-to-tissue concentration relationship, (iii) metabolism, intracellular-extracellular and extracellular-microvascular exchange, together with diffusion, determine the role of the tissue in in vivo probe behavior, and, consequently, (iv) in vitro "calibration" procedures could be useful for characterizing the probe, if properly controlled, but have limited applicability to in vivo performance. The validity of the proposed quantitative approach is illustrated by the good agreement obtained between the predictions of a model developed for tritiated water ([3]H2O) in the brain and experimental data taken from the literature for measurements in the caudoputamen of rats. The importance of metabolism and efflux to the microvasculature is illustrated by the wide variation in predicted tissue concentration profiles among [3]H2O, sucrose and dihydroxyphenylacetic acid (DOPAC).

After a single dose in mice, MPTP-N-oxide caused a dose-dependent depletion of cardiac norepinephrine which was similar although less pronounced than that caused by MPTP itself. After repeated daily doses, MPTP-N-oxide depleted cardiac norepinephrine, but did not deplete norepinephrine in the frontal cortex or dopamine in the striatum of mice. MPTP-N-oxide differed from MPTP, which depleted cardiac norepinephrine, cortical norepinephrine and striatal dopamine after repeated daily doses, but was similar to MPP+, another metabolite of MPTP, which depleted only cardiac norepinephrine. These data suggest that MPTP-N-oxide may contribute to the peripheral catecholamine-depleting effects after MPTP injection.

There are previous evidences of biochemical and functional interhemispheric asymmetry for central dopaminergic systems. The purpose of the current study was to establish an evidence of the existence of mechanisms for interhemispheric regulation of dopaminergic systems. The initial hypothesis was that to compensate the spontaneous interhemispheric asymmetry, the brain side with lower dopamine (DA) concentration show a higher DA turnover. The ratio of metabolite/neurotransmitter was computed as an index of presynaptic turnover rate. Both striatum and hippocampus showed a DOPAC/DA index higher in the brain side with a lower DA concentration. In addition, we found an inverse relation between the interhemispheric index of DA and interhemispheric index of DA turnover. Taken together the present data provide evidence of an interhemispheric regulation for the DA-innervation of both striatum and hippocampus in normal unlesioned rats.

Significant changes in monoamine levels and utilization were noted in certain brain regions of middle-aged Fisher 344 rats when compared with young adult controls. In the prefrontal cortex and septum, 3,4 dihydroxyphenylglycol (MHPG) and the MHPG/norepinephrine (NE) ratio were decreased. The septum also showed increases in dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) but there was a decrease in the DOPAC/DA ratio. The striatum showed an increase in the MPHG/NE ratio and an increase in DOPAC. The hippocampus and thalamus showed an increase in 5-hydroxyindoleacetic acid (5HIAA). This demonstrates that selected neurotransmitter systems in the brain are altered at an early stage of senescence. This could lead to ensuing neurological deficits.

Haloperidol or saline was administered to rats daily for 1, 8, 15 or 22 days. During haloperidol, but not saline administration, changes in plasma homovanillic acid (HVA) concentrations were correlated with changes in nucleus accumbens HVA. Haloperidol administration also had a significant effect on the intercorrelation of dopamine (DA) concentrations and indices of DA turnover across multiple brain areas. In particular, intercorrelations of HVA concentrations among DA terminal brain areas (i.e. striatum, nucleus accumbens, and olfactory tubercle) occurred only during haloperidol treatment.

The lateral cerebral ventricles of freely moving rats were perfused by means of chronically implanted push-pull cannulae every second day for 2 weeks. Perfusates were analyzed for metabolites of dopamine [dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA)] and of 5-hydroxytryptamine [5-hydroxyindoleacetic acid (5HIAA)] using high performance liquid chromatography and an amperometric detector. Rats received daily subcutaneous injections of haloperidol (1 mg/kg) or its vehicle. After the first injection of haloperidol the concentrations of DOPAC and HVA were markedly increased while that of 5HIAA was unchanged. Complete tolerance developed to the haloperidol-induced increased efflux of dopamine metabolites by day 9, although a higher dose of haloperidol (2 mgf/kg) on day 15 was still capable of eliciting a modest increase in the efflux of DOPAC and HVA.

Others have shown that amphetamine given to normal rats causes turning in a particular, preferred direction in most animals. We have studied the effect of training on amphetamine-induced behavioral and biochemical asymmetries in male Sprague-Dawley rats. Water deprived animals were trained to circle either in the same or opposite direction to their intrinsic bias using a sucrose water reward. Acquisition of the learned circling behavior was independent of turning preference and all animals were able to make the operant association. After training, animals given amphetamine turned in the trained direction regardless of their previous circling preference. Amphetamine-induced circling also led to increased dopamine concentrations in caudate contralateral to the trained circling direction. Therefore, intrinsic striatal lateralization is not resistant to behavioral modification and both the behavioral and biochemical asymmetries can be reversed by circling training.

Intracerebral dialysis was used to monitor the in vivo efflux of striatal dopamine (DA), homovanillic acid (HVA), dihydroxyphenylacetic acid (DOPAC) and 3-methoxytyramine (3-MT) in the pentobarbital anesthetized rat. In untreated rats, there were low levels of extra-cellular DA and 3-MT which were increased 15-fold by treatment with amphetamine. Under basal and drug-stimulated conditions, 3-MT concentrations were maintained at approximately 30% of the extracellular DA levels. These data agree with in vivo turnover estimates which indicate that 20 to 30% of DA turnover is through the 3-MT pool in the striatum. In contrast, extracellular DOPAC and HVA levels were reduced only slightly by amphetamine and with a delayed onset. Our data support the hypothesis that striatal DOPAC is an accurate index of intraneuronal DA metabolism and that 3-MT is an index of the extracellular concentration of DA.