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P Calabresi

Publications and source records attributed to P Calabresi.

At least 109 records · Page 6Linked to original sources
PubMed1997

Loss of autoreceptor function in dopaminergic neurons from dopamine D2 receptor deficient mice.

Dopamine plays a key role in the control of motor and cognitive functions through the interaction with membrane receptors. Dopamine elicits its physiological effect by interacting with receptors that belong to the seven transmembrane domain G-protein-coupled receptors family. Pharmacological and structural analyses have allowed the division of these receptors into two classes: the D1- and D2-like receptors. The D1-like subfamily comprises D1 and D5 while the D2-like is formed by D2, D3 and D4. Dopaminergic neurons arise from the ventral tegmental area and the substantia nigra. These neurons give rise to four dopaminergic pathways: the nigrostriatal, the mesolimbic, the mesocortical and tuberoinfundibular pathways. These pathways are involved in the control of movement, learning, motivation reward and hormone synthesis and release. Dysfunction in these pathways leads to neurological, psychiatric and endocrine disorders. Indeed, degeneration of the nigrostriatal pathway leads to Parkinson's disease in humans, characterized by a strong reduction of released dopamine. Thus, a fine tuning of the firing discharge of dopaminergic neurons is a key function in the regulation of dopamine mediated activities in the central nervous system. Somatodendritic dopaminergic autoreceptors of the D2-like family are responsible for such a function. However, it is still controversial whether this function could be ascribed only to one or more members of this subfamily.

Animals↗
PubMed1997

Epileptiform discharge induced by 4-aminopyridine in magnesium-free medium in neocortical neurons: physiological and pharmacological characterization.

An in vitro model of epileptiform activity was developed to study the role of excitatory and inhibitory neurotransmitters in the epileptogenesis. Intracellular recordings were obtained from rat neocortical slices exposed to 4-aminopyridine in a magnesium-free solution. Spontaneous epileptiform activity consisting of paroxysmal depolarization shifts with associated spontaneous depolarizing postsynaptic potentials were observed. The paroxysmal depolarization shifts were blocked either by D,L-2-amino-5-phosphonovalerate (50 microM), an N-methyl-D-aspartate receptor antagonist, or by 6-cyano-7-nitroquinoxaline-2.3-dione (10 microM), a non-N-methyl-D-aspartate receptor antagonist. These glutamate receptor antagonists also reduced the occurrence of spontaneous depolarizing postsynaptic potentials. Bicuculline methiodide, an antagonist of GABAA receptors, suppressed spontaneous depolarizing postsynaptic potentials, while it reduced the frequency of paroxysmal depolarization shifts and increased their duration. Hyperpolarization of the membrane potential by continuous current injection increased the frequency of paroxysmal depolarization shifts and reduced their duration, but it reduced the occurrence of spontaneous postsynaptic potentials. Paroxysmal depolarization shifts were blocked by tetrodotoxin (1 microM). The duration and the frequency of paroxysmal depolarization shift were reduced by dopamine (30-300 microM) in a dose-dependent manner. Our model suggests a different involvement of excitatory and inhibitory processes in the generation of epileptiform activity.

2-Amino-5-phosphonovalerate↗
PubMed1997

Enhancement of NMDA responses by group I metabotropic glutamate receptor activation in striatal neurones.

1. The interactions between N-methyl-D-aspartate (NMDA) and metabotropic glutamate receptors (mGluRs) were investigated in striatal slices, by utilizing intracellular recordings, both in current- and voltage-clamp mode. 2. Bath-application (50 microM) or focal application of NMDA induced a transient membrane depolarization, while in the voltage-clamp mode, NMDA (50 microM) caused a transient inward current. Following bath-application of the non-selective mGluR agonist 1S,3R-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD, 10 microM), NMDA responses were reversibly potentiated both in current (197 +/- 15% of control) and voltage-clamp experiments (200 +/- 18% of control). 3. Bath-application of the group I mGluR agonist (RS)-3,5-dihydroxyphenylglycine (3,5-DHPG, 10-300 microM) resulted in a dose-dependent potentiation of NMDA-induced membrane depolarization (up to 400 +/- 33% of control). This potentiation was either prevented by preincubation with (RS)-alpha-methyl-4-carboxyphenylglycine (RS-alpha-MCPG, 300 microM), or blocked when applied immediately after 3,5-DHPG wash-out. 4. Neither (2S,1'S,2'S)2-(2'-carboxycyclopropyl)glycine (L-CCG I, up to 100 microM) nor (2S,1'R,2'R,3'R)-2-(2,3-dicarboxycyclopropyl)-glycine (DCG-IV, 1 microM), agonists for group II mGluRs caused any change in NMDA responses. Likewise, L-serine-O-phosphate (L-SOP, 30 microM), agonist for group III mGluRs, did not affect the NMDA-induced depolarization. 5. The enhancement of the NMDA responses was mimicked by phorbol-12,13-diacetate (PDAc, 1 microM) which activates protein kinase C (PKC). The 3,5-DHPG-mediated potentiation of the NMDA-induced depolarization was prevented by preincubation with staurosporine (100 nM) or calphostin C (1 microM), antagonists of PKC. 6. Electrophysiological responses to alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptor activation were not affected by agonists for the three-classes of mGluRs. 7. The present data suggest that group I mGluRs exert a positive modulatory action on NMDA responses, probably through activation of PKC. This functional interaction in the striatum appears of crucial importance in the understanding of physiological and pathological events, such as synaptic plasticity and neuronal death, respectively.

Animals↗
PubMed1997

Prolactin response to bromocriptine in flunarizine-treated migrainous women.

Flunarizine, a calcium channel blocker, is widely used in migraine prophylaxis. Although an antidopaminergic effect has been suggested for this drug, it is unclear whether the antimigraine action of flunarizine involves the dopaminergic system. We studied the inhibitory response of prolactin to acute administration of bromocriptine, a D2 dopamine receptor agonist, before and after 1 month of treatment with flunarizine in migrainous women. Flunarizine treatment increased basal prolactin levels, but it did not reduce the inhibitory response of prolactin to acute bromocriptine administration. These findings do not support the hypothesis that flunarizine acts as a direct antagonist at the D2 dopamine receptor.

Adult↗
PubMed1997

A possible mechanism for the aglycemia-induced depression of glutamatergic excitation in the striatum.

We have studied the possible mechanisms underlying the decrease of excitatory transmission induced by glucose deprivation by using electrophysiological recordings in corticostriatal slices. Extracellular field potentials were recorded in the striatum after cortical stimulation; these potentials were progressively reduced by glucose deprivation. The reduction started 5 minutes after the onset of aglycemia. The field potential was fully suppressed after 40 minutes of glucose deprivation. After the washout of the aglycemic solution only a partial recovery was observed. Aglycemia also induced a delayed inward current during single-microelectrode voltage-clamp recordings from spiny neurons. This inward current was coupled with an increased membrane conductance. The A1 adenosine receptor antagonists, 8-cyclopentyl-1,3-dimethylxanthine (CPT, 1 micromol/L) and 1,3-dipropyl-8-cyclopentylxanthine (CPX, 300 nmol/L), significantly reduced the aglycemia-induced decrease of field potential amplitude. Moreover, in the presence of CPT and CPX, a full recovery of the field potential amplitude after the interruption of the aglycemic solution was observed. Conversely, these antagonists affected neither the inward current nor the underlying conductance increase produced by glucose deprivation. The ATP-sensitive potassium channel blockers glibenclamide (10 micromol/L) and glipizide (100 nmol/L) had no effect on the aglycemia-induced decrease of the field potential amplitude. We suggest that endogenous adenosine, but not ATP-dependent potassium channels, plays a significant role in the aglycemia-induced depression of excitatory transmission at corticostriatal synapses probably through a presynaptic mechanism. Moreover, adenosine is not involved in the postsynaptic changes induced by glucose deprivation in spiny striatal neurons.

Adenosine Triphosphate↗
PubMed1997

Astrocyte-specific expression of human T-cell lymphotropic virus type 1 (HTLV-1) Tax: induction of tumor necrosis factor alpha and susceptibility to lysis by CD8+ HTLV-1-specific cytotoxic T cells.

Human T-cell lymphotropic virus type 1 (HTLV-1) is associated with a chronic neurological disease termed HTLV-1-associated myelopathy/tropical spastic paraperesis (HAM/TSP). Although the pathogenesis of this disease remains to be elucidated, the evidence suggests that immunopathological mechanisms are involved. Since HTLV-1 tax mRNA was colocalized with glial acidic fibrillary protein, a marker for astrocytes, we developed an in vitro model to assess whether HTLV-1 infection activates astrocytes to secrete cytokines or present viral immunodominant epitopes to virus-specific T cells. Two human astrocytic glioma cell lines, U251 and U373, were transfected with the 3' portion of the HTLV-1 genome and with the HTLV-1 tax gene under astrocyte-specific promoter control. In this study, we report that Tax-expressing astrocytic glioma transfectants activate the expression of tumor necrosis factor alpha mRNA in vitro. Furthermore, these Tax-expressing glioma transfectants can serve as immunological targets for HTLV-1-specific cytotoxic T lymphocytes (CTL). We propose that these events could contribute to the neuropathology of HAM/TSP, since infected astrocytes can become a source for inflammatory cytokines upon HTLV-1 infection and serve as targets for HTLV-1-specific CTL, resulting in parenchymal damage by direct lysis and/or cytokine release.

Astrocytes↗
PubMed1997

Inhibition of growth factor mitogenicity and growth of tumor cell xenografts by a sulfonated distamycin A derivative.

Interference with growth factor-receptor interactions may have particular relevance in efforts to intervene clinically in both autocrine and paracrine aspects of malignancy. Suramin is a synthetic anticancer agent that works, in part, by blocking the binding of growth factors to their receptors. While initial clinical trials have been encouraging, its use in clinical applications is associated with significant toxicities. Suradista is a novel sulfonated distamycin derivative that is also effective at complexing and inactivating growth factors and cytokines while remaining relatively nontoxic. The goal of this study was to compare the antineoplastic properties of suramin and Suradista. To achieve this, the effects of these compounds on growth factor induced mitogenesis in normal mouse fibroblasts and human umbilical vein endothelial cells were examined, as well as their ability to inhibit the growth of NIH/3T3 cells that had been transformed by the introduction of a fibroblast growth factor (FGF) 1 coding region (residues 1-154) fused to the signal peptide of the hst/KS3 gene (sp-hst/KS3:FGF1-154). In each case, Suradista was more effective than suramin in inhibiting mitogenesis in normal cells, as well as the growth of the transformed cells. Furthermore, Suradista was also shown to be as effective as suramin at inhibiting the growth of sp-hst/KS3:FGF1-154-transformed NIH/3T3 xenografts grown in athymic nude mice when given at only 50% the dosage used for suramin (50 mg/kg for Suradista versus 100 mg/kg for suramin). In summary, these results indicate that novel compounds acting like suramin may be developed as effective antineoplastic agents and may also prove to be of clinical benefit.

3T3 Cells↗
PubMed1997

Role of dopamine receptors in the short- and long-term regulation of corticostriatal transmission.

Several studies have tried to clarify the role of different dopamine (DA) receptors in the control of membrane excitability of striatal neurons. Activation of DA receptors influences both synaptic and intrinsic membrane properties of striatal neurons. More recently it has been reported that endogenous DA plays an important role in the expression of striatal synaptic plasticity. In this review we will try to summarize and discuss the available data concerning the possible impact of the functional role of D1 and D2 receptor activation on the short- and long-term modulation of the excitatory glutamatergic corticostriatal transmission. Moreover, we will also describe the function of the striatum in the integration of glutamatergic and DAergic inputs to produce long-term changes of synaptic efficacy: long-term depression (LTD) and long-term potentiation (LTP).

Animals↗
PubMed1996

Cytokine phenotype of human autoreactive T cell clones specific for the immunodominant myelin basic protein peptide (83-99).

Experimental allergic encephalomyelitis (EAE), an animal model resembling multiple sclerosis (MS), is mediated by myelin antigen-specific CD4+ T cells secreting cytokines such as interferon-gamma (IFN-gamma), tumor necrosis factor-beta (TNF-beta), and the proinflammatory cytokine TNF-alpha-all associated with the T-helper-1 (Th1) T cell subset. Based on numerous similarities between MS and EAE, it has been postulated that Th1-like T cells are involved in the pathogenesis of MS. Production of proinflammatory cytokines such as IFN-gamma and, in particular, TNF-alpha/beta by autoreactive T cells is considered crucial for the initiation and amplification of inflammatory brain lesions and possibly also for direct myelin damage. In contrast, regulatory cytokines such as interleukin-4 (IL-4), IL-10, and IL-13, which are associated with the Th2-like phenotype, may play a role in the resolution of relapses. Although the human T cell response to myelin basic protein (MBP) is well characterized in terms of antigen specificity, HLA restriction, and T cell-receptor (TCR) usage, little is known about the cytokine pattern of these autoreactive T cells. To gain such information, conditions for studying cytokine secretion by human autoreactive T cell clones (TCC) were established. The cytokine secretion profile of human autoreactive CD4+ TCC, specific for myelin basic protein peptide (83-89) [MBP(83-99)], a candidate autoantigen in MS, was investigated. Our results show that TCC cytokine production in long-term culture was stable. In addition, the correlation of various cytokines within specific TCC revealed differences compared to murine T cells. The comparison of 30 human MBP (83-99)-specific TCC demonstrated heterogeneity in cytokine secretion, with a continuum between Th1- and Th2-like cells rather than distinct Th1 or Th2 subsets. These data are important for further investigation of the potential role of cytokines in the inflammatory process of MS, and provide a powerful tool to investigate therapeutic interventions with respect to their influence on cytokine secretion of autoreactive T cells.

Amino Acid Sequence↗
PubMed1996

The modulation of calcium currents by the activation of mGluRs. Functional implications.

Glutamatergic transmission in the central nervous system (CNS) is mediated by ionotropic, ligand-gated receptors (iGluRs), and metabotropic receptors (mGluRs). mGluRs are coupled to GTP-binding regulatory proteins (G-proteins) and modulate different second messenger pathways. Multiple effects have been described following their activation; among others, regulation of fast synaptic transmission, changes in synaptic plasticity, and modification of the threshold for seizure generation. Some of the major roles played by the activation of mGluRs might depend on the modulation of high-voltage-activated (HVA) calcium (Ca2+) currents. Some HVA Ca2+ channels (N-, P-, and Q-type channels) are signaling components at most presynaptic active zones. Their mGluR-mediated inhibition reduces synaptic transmission. The interference, by agonists at mGluRs, on L-type channels might affect the repetitive neuronal firing behavior and the integration of complex events at the somatic level. In addition, the mGluR-mediated effects on voltage-gated Ca2+ signals have been suggested to strongly influence neurotoxicity. Rather different coupling mechanisms underlie the relation between mGluRs and Ca2+ currents: Together with a fast, membrane-delimited mechanism of action, much slower responses, involving intracellular second messengers, have also been postulated. In the recent past, the relative paucity of selective agonists and antagonists for the different subclasses of mGluRs had hampered the clear definition of the roles of mGluRs in brain function. However, the recent availability of new pharmacological tools is promising to provide a better understanding of the neuronal functions related to different mGluR subtypes. The analysis of the mGluR-mediated modulation of Ca2+ conductances will probably offer new insights into the characterization of synaptic transmission and the development of neuroprotective agents.

Animals↗
PubMed1996

The corticostriatal projection: from synaptic plasticity to dysfunctions of the basal ganglia.

Corticostriatal transmission has an important function in the regulation of the neuronal activity of the basal ganglia. The firing activity of corticostriatal neurones excites striatal cells via the release of glutamate. Presynaptic receptors that are located on corticostriatal terminals and that regulate the release of glutamate in the striatum have been postulated for dopamine and glutamate. Activation of these receptors may exert a negative feed-back on the striatal release of glutamate. High-frequency activation of corticostriatal fibres causes either long-term depression or long-term potentiation of excitatory transmission depending on the subclass of glutamate receptor that is activated. These forms of synaptic plasticity could be involved in motor learning. Alterations in striatal synaptic plasticity might be implicated in Parkinson's disease and Huntington's disease.

Animals↗
PubMed1996

Characterization of a barium-sensitive outward current following glutamate application on rat midbrain dopaminergic cells.

Using intracellular electrophysiological recordings in dopaminergic (principal) neurons of the rat mesencephalon maintained in vitro, we studied a postexcitatory amino acid response (PEAAR). Under current-clamp mode, bath application of glutamate produced a depolarization that was followed by a hyperpolarization when the perfusion of the excitatory amino acid was discontinued. Under single-microelectrode voltage-clamp mode, an outward current followed the glutamate-induced inward current. The PEAAR was associated with an increase in membrane conductance and reversed polarity at about-85 mV (2.5 mM extracellular K+). The null potential for the PEAAR was independent of the intracellular loading of chloride ions and was shifted towards less negative values (approximately 23 mV) by increasing extracellular K+ from 2.5 to 8.5 mM. The PEAAR was present in neurons treated with tetraethylammonium (5-10 mM), apamin (1 microM) or glibenclamide (1-300 microM). However, it was strongly depressed or blocked by extracellular barium (300 microM to 1 mM), by low-calcium (0.5 mM) plus cadmium (100 microM) or magnesium (10 mM), and by low-sodium solutions. An outward response was also generated after an inward current induced by the perfusion of the specific agonists for the ionotropic excitatory amino acid receptors NMDA, alpha-amino-3-hydroxy-5-methyl-4-isoxazole (AMPA) and kainate. The PEAAR was not affected by tetrodotoxin (1 microM), saclofen (100-300 microM), bicuculline (30 microM), sulpiride (1 microM) or strychnine (1 microM). In addition, the inhibition of the ATP-dependent Na(+)-K+ pump by ouabain and strophanthidin (1-10 microM) prolonged the glutamate-induced membrane depolarization/inward current while the subsequent PEAAR was reduced or not observed. Our data indicate that the PEAAR mainly results from the activation of a barium-sensitive potassium current. This response might limit the excitatory and eventually neurotoxic effects of glutamate.

Animals↗
PubMed1996

Electrophysiological effects of monoamine oxidase inhibition on rat midbrain dopaminergic neurones: an in vitro study.

1 The effects of the inhibition of monoamine oxidase (MAO) type A and B have been evaluated on the spontaneous firing activity of the dopaminergic (principal) neurones of the rat midbrain intracellularly recorded from a slice preparation. 2 The non-specific MAO inhibitor, pargyline, superfused at a concentration of 10-100 microM, decreased or abolished the spontaneous firing discharge of the principal neurons in the subtantia nigra pars compacta and ventral tegmental area. This effect had a slow onset and appeared to be sustained. 3 The administration of the dopamine D2/3 receptor antagonist, sulpiride (100-300 nM), antagonized the pargyline-induced effect, while the superfusion of the dopamine D1 receptor antagonist, SCH 23390 (1-3 microM) did not counteract the induced inhibition of the firing rate. 4 The inhibitor for the MAO A, clorgyline (30-100 microM), reduced the firing rate of the dopaminergic neurones. A similar depressant effect was also observed when a MAO B inhibitor, deprenyl (30-100 microM), was applied. Lower concentrations of both drugs (300 nM-10 microM) did not produce consistent effects on neuronal discharge. 5 Our data suggest that only the blockade of both types of MAO enzymes favours the inhibitory action of endogenous dopamine on somato-dendritic D2/3 autoreceptors.

Animals↗
PubMed1996

A field potential analysis on the effects of lamotrigine, GP 47779, and felbamate in neocortical slices.

We studied the action of the new antiepileptic drugs lamotrigine (LTG), GP 47779 (the active metabolite of oxcarbazepine), and felbamate (FBM) on stimulus-evoked field potentials recorded from rat prefrontal and frontal cortical slices. In the presence of physiologic concentrations of extracellular magnesium (1.2 mM) the field potential amplitude was not affected by the N-methyl-D-aspartate (NMDA) glutamate receptor antagonist, 2-amino-5-phosphonovalerate (APV), while it was blocked by the non-NMDA glutamate receptor antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). When magnesium was removed from the bathing medium, there was a significant NMDA-mediated component of the field potential. LTG and GP 47779 decreased, in a dose-dependent manner, the field potential amplitude under both experimental conditions. FBM caused a dose-related decrease of the field potential amplitude only in the absence of external magnesium, suggesting a selective interaction with an NMDA-mediated component of this potential. These findings indicate that the reduction of cortical excitatory transmission might represent a common target for new antiepileptic drugs.

Animals↗
PubMed1996

Felbamate inhibits dihydropyridine-sensitive calcium channels in central neurons.

The effect of the antiepileptic drug felbamate (FBM) on high-voltage-activated Ca++ currents was studied in cortical and neostriatal neurons acutely isolated from adult rats. Patch-clamp recordings in the whole-cell configuration were performed. Ba++ ions as the charge carrier for Ca++ channels were used. In pyramidal cortical cells, FBM dose-dependently reduced high-voltage-activated Ca++ currents in all the tested neurons. At concentrations of 30 to 100 nM, FBM already produced a significant inhibition of high-voltage-activated Ca++ currents (-6/-15%). At saturating concentrations (1-3 microM), FBM-mediated inhibition averaged 44%. The responses were fully reversible. The dose-response curves revealed IC50 of 504 nM. In striatal neurons, FBM decreased the same conductances by about 28%; the threshold dose was 1 to 2 microM, with an IC50 of 18.7 microM. In both structures, the observed inhibitions were unaffected by omega-conotoxin GVIA and omega-agatoxin IVA, suggesting that N-like channels and P-Like channels were not involved in the FBM-mediated responses. In addition, when omega-conotoxin GVIA and omega-agatoxin IVA (100 nM) were coapplied, the FBM-mediated inhibition on the remaining Ca++ currents averaged 87%. The FBM responses were occluded by micromolar concentrations of nifedipine, supporting a direct interference with dihydropyridine-sensitive channels. It is concluded that the described effect of FBM might represent an efficacious mechanism for either controlling spike discharge from epileptic foci or protecting neurons from excessive Ca++ loading. In both cases, FBM would act as a broad spectrum neuroprotective agent.

Animals↗
PubMed1995

Actions of glycine on non-dopaminergic neurons of the rat substantia nigra.

The effects of glycine on non-dopaminergic cells in rat substantia nigra pars compacta and pars reticulata maintained in vitro were investigated using intracellular recording techniques. Glycine, superfused at a concentration between 30 microM and 1 mM, reversibly blocked the spontaneous firing of these neurons. The inhibition of firing discharge was associated with a hyperpolarization of the membrane (potassium acetate-filled electrodes) and an increase in conductance. Under voltage-clamp experiments (holding potential between -57 and -65 mV), glycine produced an outward response which reversed polarity at about -74 mV. However, when the recording electrodes were filled with KCl, the glycinergic response was mainly depolarizing/inward and reversed at about -43 mV. Thus, it appeared to be due to an increase in chloride permeability. Furthermore, the effects of glycine were reversibly antagonized by strychnine (between 300 nM and 1 microM). Our findings demonstrate that glycine is a potent inhibitory agent on non-dopaminergic cells of the substantia pars compacta and par reticulata that acts by activating strychnine-sensitive receptors.

Animals↗