Differential effects of cocaine-induced seizures and lethality on M(1)-like muscarinic and dopaminergic D (1)- and D (2)-like binding receptors in mice brain.

This work was designed to study the changes produced by cocaine-induced seizures and lethality on dopaminergic D(1)- and D(2)-like receptors, muscarinic M(1)-like binding sites, as well as acetylcholinesterase activity in mice prefrontal cortex (PFC) and striatum (ST). Binding assays were performed in brain homogenates from the PFC and ST and ligands used were [(3)H]-N-methylscopolamine, [(3)H]-NMS (in the presence of carbachol), [(3)H]-SCH 23390 and [(3)H]-spiroperidol (in presence of mianserin), for muscarinic (M(1)-like), D(1)- and D(2)-like receptors, respectively. Brain acetylcholinesterase (AChE) activity was also determined in these brain areas. Cocaine-induced SE decreased [(3)H]-SCH 23390 binding in both ST and PFC areas. A decrease in [(3)H]-NMS binding and an increase in [(3)H]-spiroperidol binding in PFC was also observed. Cocaine-induced lethality increased [(3)H]-spiroperidol binding in both areas and decreased [(3)H]-NMS binding only in PFC, while no difference was seen in [(3)H]-SCH 23390 binding. Neither SE, nor lethality altered [(3)H]-NMS binding in ST. AChE activity increased after SE in ST while after death the increase occurred in both PFC and ST. In conclusion, cocaine-induced SE and lethality produces differential changes in brain cholinergic and dopaminergic receptors, depending on the brain area studied suggesting an extensive and complex involvement of these with cocaine toxicity in central nervous system.

Behavioral and neurochemical alterations after lithium-pilocarpine administration in young and adult rats: a comparative study.

Pilocarpine and lithium-pilocarpine can induce seizures and brain damage in adult rats. However, manifestation of cerebral lesions seems to be an age-related phenomenon suggesting that maturational states of neurocircuitry may be involved. We have studied behavior changes, cerebral histopathology, and muscarinic and dopaminergic receptors density in rodents subjected to lithium-pilocarpine treatment. Wistar rats, at two different ages (21 days and 2 months), were treated with pilocarpine (15 mg/kg, SC), lithium (3 mEq/kg, IP), atropine (50 mg/kg, IP) and the combination of lithium to pilocarpine. Histopathologic studies showed that younger animals were more resistant to the development of cerebral changes and there was a preferential involvement of the striatum (Wilcoxon p = 0.02) as opposed to more generalized areas in adult animals such as hippocampus and neocortex. Lithium treatment induced an upregulation of muscarinic receptors at both ages, and this effect was reversed in younger animals after pilocarpine administration. Lithium also induced an upregulation of dopaminergic receptors in the striatum at both ages (p < 0.05), and this effect was not reversed after pilocarpine administration. Our data confirm that young animals show less brain damage after lithium-pilocarpine, and main alterations in dopaminergic receptors density occur in young and older animals after treatment with lithium and lithium combined to a low dose of pilocarpine.

[3H]raclopride and [3H]spiroperidol binding to retinal membranes of the teleost Eugerres plumieri: effect of light and dark adaptation.

Monoamine and metabolites were determined in the retina of the teleost Eugerres plumieri after dark and light adaptation. Dopamine, homovanillic acid and 3,4-dihydroxyphenylacetic acid increased after light exposure. The results indicate an increase in the turnover rate of dopamine due to light exposure. Dopamine D2 receptors were studied by determining the binding parameters of [3H]spiroperidol and [3H]raclopride to retinal membranes. The results were best fitted to a two-site model, where the high-affinity site may correspond to D2 receptors and the low-affinity site could be D4 receptors, which have been recently described in the retina, although further research is needed to confirm this suggestion. The number of sites labeled with [3H]spiroperidol was lower than with [3H]raclopride. This may indicate the existence of monomer and dimer conformations of D2-like receptors in the retina, as has been shown in the brain. Light exposure increased the number of sites labeled with both ligands. Since D2 receptors are known to modulate the production of melatonin, the augmentation in the capacity of these receptors could contribute to the reduction of melantonin during light exposure.

Effect of a corn diet during development on [3H]-spiperone binding in the brain of rats at the perinatal stage.

A model of undernutrition based on feeding with a corn-diet (tryptophan deficient) was used to study maturation and physiology of the rat brain 5-HTergic system. Previously, using that model, we observed a decrease in 5-HT uptake in rat brain slices, assuming that some compensatory mechanisms could be implicated at the synaptic level on those conditions. Body, brain weight, and total [3H]-spiperone binding were recorded at two fetal ages and newborn pups from mothers were fed with a corn-diet and with a corn-lysine diet. Significant decreases in body weight and in [3H]-spiperone binding were observed at all ages studied in the corn-based diet groups with respect to controls at the first postnatal day of age, whereas at the 18th gestation day, the [3H]-spiperone binding was higher in the protein restricted and corn-lysine supplemented group than in the control group. Results suggested that expression of 5-HT receptors and possibly their synthesis may be limited at the synaptic level on undernutrition conditions.

[The binding of 3H-spiperone to lymphocytes in patients with schizophrenia and first-degree relatives: in search of a genetic marker]. / La unión de la 3H-espiperona a linfocitos en pacientes con esquizofrenia y familiares de primer grado: en busca de un marcador genético.

3H-spiperone binding of lymphocytes has been studied and proposed to be a useful vulnerability marker for certain subtypes of schizophrenia. The binding of 3H-spiperone into lymphocytes was studied in 22 schizophrenics, 19 unaffected relatives and 8 healthy subjects. Binding assay was performed, incubating lymphocytes with 0.04 to 0.4 mM of 3H-spiperone. Non-specific binding was measured with 10 mM haloperidol. At the end the lymphocytes were homogenized. The results show a significant (p < 0.01) lower affinity (high Kd values) in drug naive schizophrenics compared with the controls. No differences were found in the binding capacity (Bmax) of both schizophrenic groups compared with control subjects. The results cannot offer us a reliable biological marker for schizophrenia. The increase of the number of subjects is suggested.

Daily variation in plasma concentration of fencamfamine and striatal dopamine receptors in rats.

Fencamfamine (FCF) is a psychostimulant drug classified as an indirect dopamine agonist. In the present study we evaluated the daily variation in plasma FCF concentration and in striatal dopamine receptors. Adult male Wistar rats (250-300 g) maintained on a 12-h light/12-h dark cycle (lights on at 07:00 h) were used. Rats received FCF (10.0 mg/kg, ip) at 09:00, 15:00, 21:00 or 03:00 h and blood samples were collected 30 (N = 6) or 60 (N = 6) min after the injections. Plasma FCF was measured by gas chromatography using an electron capture detector. Two-way ANOVA showed significant differences in FCF concentration when blood samples were collected 30 min after the injection, and the highest value was obtained following injection at 21:00 h. Moreover, at 15:00, 21:00 and 03:00 h, plasma FCF levels were significantly lower 60 min after injection when compared to the 30-min interval. Two other groups of rats (N = 6) were decapitated at 09:00 or 21:00 h and the striata were dissected for the binding assays. The Bmax for [3H]-spiroperidol binding to striatal membranes was higher at 21:00 h, without changes in affinity constant (Kd). In conclusion, plasma FCF levels and dopamine receptors undergo daily variation, a phenomenon that should be considered to explain the circadian time-dependent effects of FCF.

Daily variation in plasma concentration of fencamfamine and striatal dopamine receptors in rats

Fencamfamine (FCF) is a psychostimulant drug classified as an indirect dopamine agonist. In the present study we evaluated the daily variation in plasma FCF concentration and in striatal dopamine receptors. Adult male Wistar rats (250-300 g) maintained on a 12-h light/12-h dark cycle (lights on at 07:00 h) were used. Rats received FCF (10.0 mg/kg, ip) at 09:00, 15:00, 21:00 or 03:00 h and blood samples were collected 30 (N = 6) or 60 (N = 6) min after the injections. Plasma FCF was measured by gas chromatography using an electron capture detector. Two-way ANOVA showed significant differences in FCF concentration when blood samples were collected 30 min after the injection, and the highest value was obtained following injection at 21:00 h. Moreover, at 15:00, 21:00 and 03:00 h, plasma FCF levels were significantly lower 60 min after injection when compared to the 30-min interval. Two other groups of rats (N = 6) were decapitated at 09:00 or 21:00 h and the striata were dissected for the binding assays. The Bmax for [3H]-spiroperidol binding to striatal membranes was higher at 21:00 h, without changes in affinity constant (Kd). In conclusion, plasma FCF levels and dopamine receptors undergo daily variation, a phenomenon that should be considered to explain the circadian time-dependent effects of FCF

Autoradiographic analysis of D1 and D2 dopaminergic receptors in rat brain after paradoxical sleep deprivation.

Previous work had shown that paradoxical sleep deprivation (PSD) results in potentiation of several apomorphine-induced behaviors, leading to the suggestion that PSD induces an upregulation of brain dopamine receptors. In this study, quantitative receptor autoradiography was used to verify whether PSD does, in fact, induce alterations in D1 or D2 receptor binding, and to investigate the regional brain specificity of such effects. After 96 h of PSD, [3H]SCH-23390 binding to D1 receptors was examined in 30 different brain areas of 10 experimental and 10 cage control rats. [3H]Spiperone was used to label D2 sites in adjacent tissue sections. Results revealed a 39% increase in [3H]SCH-23390 binding in the entorhinal cortex of PSD rats (p < 0.05), but no other changes in any of the remaining 29 brain areas examined. In contrast, [3H]spiperone binding was significantly elevated in the n. accumbens (+45%) and in all subregions of the caudate-putamen (range: +13% to +23%). These results, thus, provide evidence that PSD increases D2 but not D1 receptor binding in brain. The present results also suggest that upregulated D2 receptors can account for the previously reported changes in apomorphine-induced behaviors after PSD.

Adaptive mechanisms of striatal D1 and D2 dopamine receptors in response to a prolonged reserpine treatment in mice.

Mice receiving reserpine (1 mg/kg/day) during 5 days develop behavioral supersensitivity. To study the possible molecular correlates of these adaptive changes we compared binding parameters of D1 and D2 receptors and adenylate cyclase activity in striata from normal and reserpinized mice. Saturation curves using [3H]SCH 23390 showed no changes in maximum binding capacity (Bmax) or Kd of striatal D1 receptors taken from control or 5 days reserpine-treated mice. However, [3H]spiperone saturation curves showed a 31% increase in D2 receptors Bmax with no changes in Kd. Dopamine competition of [3H]SCH 23390 and [3H]spiperone binding in mouse striatum was also performed. Analysis of data by LIGAND showed that dopamine recognizes two subpopulations for D1 and for D2 receptors. The proportion of receptors in the high affinity state (D1high and D2high) were increased in reserpine-treated animals. The addition of 100 microM GTP produced a complete conversion of D1high and D2high receptors into their low-affinity states in striata from control and reserpinized mice. Five days of reserpine treatment increased basal adenylate cyclase activity of mouse striatum in the presence of Mn++ or Mg++ ions. Concentration curves with dopamine, NaF or forskolin revealed shifts to the left and higher maximum responses without changes in EC50 values in striata from reserpinized mice. Thus, a prolonged reserpine treatment produces marked changes in D1 and D2 receptors increasing the proportion of high affinity state subpopulations and the total Bmax of D2 receptors. Also, dopamine function may be enhanced through an increment of the catalytic component of striatal adenylate cyclase.

Striatal dopamine receptor supersensitivity after long-term haloperidol treatment of hypophysectomized rats.

Dopamine (DA) receptor sensitivity was studied after long-term treatment with haloperidol (0.5 and 3.0 mg/kg, ip, single daily dose) or saline in hypophysectomized and intact rats. Haloperidol treatment for seven days produced a 25 to 125% increase in [3H]-spiroperidol binding to striatal DA receptors in a dose-dependent fashion. The increase in binding sites (Bmax) was similar in both hypophysectomized and intact rats when compared to controls. The present results show that hypophysectomy does not effect the supersensitivity of striatal DA receptors induced by long-term haloperidol treatment.

(3H) spiroperidol binding decreases in brains of rats infected with Venezuelan equine encephalomyelitis virus.

After i.p. inoculation with the Guajira strain of Venezuelan equine encephalomyelitis virus a significant decrease in the density of (3H) spiroperidol binding sites in the striatum, midbrain and frontal cortex was observed. No changes in the affinity of the receptors could be demonstrated. This finding is compatible with neuronal degeneration caused by the viral infection.