ABSTRACT
Clinical trial sponsors have ethical obligations to register protocols, report study results and comply with applicable legal requirements. To evaluate public commitments to trial disclosure and rates of disclosure by members and non-members of the European Federation of Pharmaceutical Industries and Associations (EFPIA) and/or the Pharmaceutical Research and Manufacturers of America (PhRMA). Websites of the top 50 biopharmaceutical companies by 2015 sales were searched for statements relating to trial data disclosure. Disclosure of trial results completed by biopharmaceutical industry and non-industry sponsors of at least 30 trials (2006-2015) was assessed using TrialsTracker. Among the top 50 companies, 30 were EFPIA/PhRMA members and 20 were non-members, of which 26 and none, respectively, had a statement on their website committing to the disclosure of trials data. Of 29 377 trials in TrialsTracker, 9511 were industry sponsored (69 companies) and 19 866 were non-industry sponsored (254 institutions). The overall mean disclosure rate was 55%, with higher rates for industry (74%) than for non-industry sponsors (46%). Of the 30 companies within the top 50 with data in TrialsTracker, the mean disclosure rate was 76% (77% for EFPIA/PhRMA members [n=25] vs 67% for non-members [n=5]). Most of the top 50 biopharmaceutical companies have publicly committed to the disclosure of trial data. Industry sponsors have responded to the ethical and legal demands of trial disclosure by disclosing three quarters of their trials compared with less than half for non-industry sponsors. Further improvements in clinical trial disclosure are needed.
Subject(s)
Access to Information , Clinical Trials as Topic , Disclosure , Drug Industry , Clinical Trials as Topic/standards , Drug Industry/organization & administration , Humans , RegistriesABSTRACT
RATIONALE: Prefrontal cortex (PFC)-dependent executive function is disrupted in a range of psychiatric disorders and can be modelled in non-human primates and rodents using attentional set-shifting paradigms. There are few current pharmacological strategies for enhancing attentional set shifting, although the PFC is rich in relevant neurotransmitter targets, including 5-hydroxytryptamine (5-HT). Although 5-HT depletion studies do not support a role for 5-HT in attentional set shifting, the effect of 5-HT activation using specific receptor agonists has not been tested. OBJECTIVES AND METHODS: This study investigated the effect of a novel, selective 5-HT(6) receptor agonist, WAY181187, in a rat model of PFC-dependent extra-dimensional (ED) attentional set shifting. The effect of this agent on immediate early gene expression in the medial PFC and other regions was also examined. RESULTS: Compared to vehicle-injected controls, WAY181187 facilitated ED set shifting but did not change other non-ED phases of the task (including intra-dimensional set shifting and reversal). This effect was blocked by the selective 5-HT(6) antagonist SB399885, which alone had no effect. WAY181187 enhanced ED set shifting even when administered after the attentional set had been acquired, thereby ruling out impairments in attentional set formation. In separate experiments, at a dose that increased ED set shifting, WAY181187 increased Fos-like immunoreactivity in the medial PFC in a SB399885-sensitive manner, suggesting a 5-HT(6) receptor-mediated activation of this region. CONCLUSIONS: Through use of a novel 5-HT agonist, these experiments reveal a previously unrecognised role for 5-HT activation in PFC-dependent executive function, mediated by 5-HT(6) receptor activation.
Subject(s)
Attention/drug effects , Receptors, Serotonin/physiology , Serotonin Receptor Agonists/pharmacology , Thiazoles/pharmacology , Tryptamines/pharmacology , Animals , Behavior, Animal/drug effects , Dose-Response Relationship, Drug , Gene Expression Regulation/drug effects , Male , Piperazines/pharmacology , Prefrontal Cortex/drug effects , Prefrontal Cortex/metabolism , Rats , Receptors, Serotonin/drug effects , Reversal Learning/drug effects , Serotonin Receptor Agonists/administration & dosage , Set, Psychology , Sulfonamides/pharmacology , Thiazoles/administration & dosage , Tryptamines/administration & dosageABSTRACT
Clinical evaluation of tachykinin NK(3) receptor antagonists has provided support for the therapeutic utility of this target in schizophrenia. However, these studies have not been entirely conclusive, possibly because of the pharmacokinetic limitations of these molecules. In the search for tachykinin NK(3) receptor antagonists with improved properties, we have discovered GSK172981 and GSK256471. Both compounds demonstrated high affinity for recombinant human (pK(i) values 7.7 and 8.9, respectively) and native guinea pig (pK(i) values 7.8 and 8.4, respectively) tachykinin NK(3) receptors. In vitro functional evaluations revealed GSK172981 to be a competitive antagonist (pA(2)=7.2) at cloned human tachykinin NK(3) receptor whereas GSK256471 diminished the neurokinin B-induced E(max) response, indicative of non-surmountable antagonist pharmacology (pA(2)=9.2). GSK172981 also exhibited a competitive profile in antagonizing neurokinin B-stimulated neuronal activity recorded from the guinea pig medial habenula slices (apparent pK(B)=8.1), whilst GSK256471 abolished the agonist-induced response. Central nervous system penetration by GSK172981 and GSK256471 was indicated by dose-dependent ex vivo tachykinin NK(3) receptor occupancy in medial prefrontal cortex (ED(50) values of 0.8 and 0.9 mg/kg, i.p., respectively) and the dose-dependent attenuation of agonist-induced "wet dog shake" behaviours in guinea pigs. Finally, in vivo microdialysis studies demonstrated that acute GSK172981 (30 mg/kg, i.p.) and GSK256471 (1mg/kg, i.p.) attenuated haloperidol-induced increases in extracellular dopamine in the guinea pig nucleus accumbens. Taken together, these in vitro and in vivo characterisations of the tachykinin NK(3) receptor antagonists GSK172981 and GSK256471 support their potential utility in the treatment of schizophrenia.
Subject(s)
Aminoquinolines/metabolism , Aminoquinolines/pharmacology , Brain/drug effects , Brain/metabolism , Quinolines/metabolism , Quinolines/pharmacology , Receptors, Tachykinin/antagonists & inhibitors , Aminoquinolines/pharmacokinetics , Animals , Behavior, Animal/drug effects , Brain/cytology , Brain/physiology , Cerebral Cortex/drug effects , Cerebral Cortex/metabolism , Cloning, Molecular , Dopamine/metabolism , Extracellular Space/drug effects , Extracellular Space/metabolism , Guinea Pigs , Habenula/cytology , Haloperidol/pharmacology , Humans , Inositol Phosphates/metabolism , Male , Microdialysis , Neurokinin B/pharmacology , Neurons/cytology , Neurons/drug effects , Nucleus Accumbens/drug effects , Nucleus Accumbens/metabolism , Peptide Fragments/pharmacology , Peptides/chemistry , Quinolines/pharmacokinetics , Rats , Receptors, Tachykinin/genetics , Receptors, Tachykinin/metabolism , Substance P/analogs & derivatives , Substance P/pharmacologyABSTRACT
A hypofunction of the N-methyl-D-aspartate (NMDA) receptor has been implicated in the pathophysiology of schizophrenia. Compelling evidence of altered NMDA receptor subunit expression in the schizophrenic brain has not, however, so far emerged. Rats reared in isolation exhibit several characteristics, including disturbed sensory gating, which resemble those seen in schizophrenia. To explore the possibility that NMDA receptor dysfunction may contribute to the behavioral and neurochemical consequences of rearing rats in isolation, we compared NMDA receptor subunit expression in brains of rats which were housed in isolation and which displayed a deficit in prepulse inhibition of the acoustic startle response with that of socially housed controls. An initial microarray analysis revealed a 1.26-fold increase in NR2A transcript in the prefrontal cortex, but not in the nucleus accumbens, of rats reared in isolation compared with those housed socially. In contrast, NR1, NR2B, NR2C, NR2D, NR3A, and NR3B subunit expression was unchanged in either brain area. In a second cohort of animals, in situ hybridization revealed increased NR2A mRNA expression in the medial prefrontal cortex, an observation that was substantiated by increased [(3)H]CGP39653 binding suggesting that NR2A receptor subunit protein expression was also elevated in the medial prefrontal cortex of the same animals. No changes in expression of NR1 or NR2B subunits were observed at both mRNA and protein level. Altered NR2A subunit expression in the medial prefrontal cortex of rats reared in isolation suggests that NMDA receptor dysfunction may contribute to the underlying pathophysiology of this preclinical model of aspects of schizophrenia.
Subject(s)
Gene Expression Regulation/physiology , Prefrontal Cortex/metabolism , Receptors, N-Methyl-D-Aspartate/metabolism , Social Isolation , 2-Amino-5-phosphonovalerate/analogs & derivatives , 2-Amino-5-phosphonovalerate/metabolism , Acoustic Stimulation/adverse effects , Animals , Animals, Newborn , Gene Expression Profiling/methods , Indoles/metabolism , Male , Oligonucleotide Array Sequence Analysis/methods , Protein Binding/physiology , Radioligand Assay/methods , Rats , Receptors, N-Methyl-D-Aspartate/genetics , Reflex, Startle/physiology , Tritium/metabolismABSTRACT
OBJECTIVES: To test the novel nonimidazole histamine H3 receptor antagonist 5-[(3-cyclobutyl-2,3,4,5-tetrahydro-1H-3-benzazapin-7-yl)oxy]-N-methyl-2-pyrazinecarboxamide (GSK207040) in a series of behavioral and neurochemical paradigms designed to evaluate its antipsychotic potential. MATERIALS AND METHODS: Acute orally administered GSK207040 was investigated for its capacity to reverse a 24-h-induced deficit in novel object recognition memory, deficits in prepulse inhibition (PPI) induced by isolation rearing, and hyperlocomotor activity induced by amphetamine. The acute neurochemical effects of GSK207040 were explored by analyzing rat anterior cingulate cortex microdialysates for levels of dopamine, noradrenaline, and acetylcholine and by c-fos immunohistochemistry. The potential for interaction with the antipsychotic dopamine D2 receptor antagonist haloperidol was explored behaviorally (spontaneous locomotor activity and catalepsy), biochemically (plasma prolactin), and via ex vivo receptor occupancy determinations. RESULTS: GSK207040 significantly enhanced object recognition memory (3 mg/kg) and attenuated isolation rearing-induced deficits in PPI (1.0 and 3.2 mg/kg) but did not reverse amphetamine-induced increases in locomotor activity. There was no evidence of an interaction of GSK207040 with haloperidol. GSK207040 (3.2 mg/kg) raised extracellular concentrations of dopamine, noradrenaline, and acetylcholine in the anterior cingulate cortex and c-fos expression in the core of the nucleus accumbens was increased at doses of 3.2 and 10.0 mg/kg. CONCLUSIONS: The behavioral and neurochemical profile of GSK207040 supports the potential of histamine H3 receptor antagonism to treat the cognitive and sensory gating deficits of schizophrenia. However, the failure of GSK207040 to reverse amphetamine-induced locomotor hyperactivity suggests that the therapeutic utility of histamine H(3) receptor antagonism versus positive symptoms is less likely, at least following acute administration.
Subject(s)
Antipsychotic Agents/pharmacology , Benzazepines/pharmacology , Histamine Antagonists/pharmacology , Pyrazines/pharmacology , Schizophrenia/drug therapy , Administration, Oral , Amphetamine/pharmacology , Animals , Antipsychotic Agents/administration & dosage , Behavior, Animal/drug effects , Benzazepines/administration & dosage , Dose-Response Relationship, Drug , Histamine Antagonists/administration & dosage , Hyperkinesis/chemically induced , Hyperkinesis/prevention & control , Male , Memory/drug effects , Pyrazines/administration & dosage , Rats , Rats, Sprague-Dawley , Receptors, Histamine H3/drug effects , Recognition, Psychology/drug effects , Schizophrenia/physiopathology , Social Isolation/psychologyABSTRACT
Chandelier neurons are a subset of parvalbumin containing cortical interneurons characterised by their preferential targeting of the axon initial segments of pyramidal neurons. They have been the focus of recent interest after evidence that the arrays of boutons are reduced in the prefrontal cortex of schizophrenic patients, post mortem. Since one chandelier neuron may innervate the axon initial segments of several hundred pyramidal neurons, it is hypothesized that their special connectivity might facilitate synchronisation of cortical outputs and play a key role in working memory. Disruption in their function is therefore thought to play a potentially important role in cortically associated symptoms of schizophrenia. Using the isolation rearing animal model of schizophrenia, we examined immunolabelling for GABA-transporter 1, a marker of chandelier cartridges. We show that the numbers of arrays of chandelier axons are reduced by 36% in the ventral prelimbic cortex of isolation-reared rats, compared with their socially-housed litter mates. This mimics findings in the PFC of schizophrenic patients where GAT-1-positive cartridges are reduced by 40% and is the first study to demonstrate changes in chandelier cartridges in an animal model of schizophrenia.
Subject(s)
Axons/pathology , GABA Plasma Membrane Transport Proteins/metabolism , Interneurons/pathology , Prefrontal Cortex/pathology , Social Isolation/psychology , gamma-Aminobutyric Acid/metabolism , Animals , Axons/metabolism , Behavior, Animal/physiology , Cell Differentiation/physiology , Disease Models, Animal , Environment, Controlled , Immunohistochemistry , Interneurons/metabolism , Male , Neural Inhibition/physiology , Prefrontal Cortex/growth & development , Prefrontal Cortex/metabolism , Rats , Schizophrenia/metabolism , Schizophrenia/pathology , Schizophrenia/physiopathology , Sensory Deprivation/physiologyABSTRACT
Neurokinin-3 (NK3) receptors are concentrated in forebrain and basal ganglia structures within the mammalian CNS. This distribution, together with the modulatory influence of NK3 receptors on monoaminergic neurotransmission, has led to the hypothesis that NK3 receptor antagonists may have therapeutic efficacy in the treatment of psychiatric disorders. Here we describe the in vitro and in vivo characterization of the highly selective NK3 receptor antagonist talnetant (SB-223412). Talnetant has high affinity for recombinant human NK3 receptors (pKi 8.7) and demonstrates selectivity over other neurokinin receptors (pKi NK2 = 6.6 and NK1<4). In native tissue-binding studies, talnetant displayed high affinity for the guinea pig NK3 receptor (pKi 8.5). Functionally, talnetant competitively antagonized neurokinin B (NKB)-induced responses at the human recombinant receptor in both calcium and phosphoinositol second messenger assay systems (pA2 of 8.1 and 7.7, respectively). In guinea pig brain slices, talnetant antagonized NKB-induced increases in neuronal firing in the medial habenula (pKB = 7.9) and senktide-induced increases in neuronal firing in the substantia nigra pars compacta (pKB = 7.7) with no diminution of maximal agonist efficacy, suggesting competitive antagonism at native NK3 receptors. Talnetant (3-30 mg/kg i.p.) significantly attenuated senktide-induced 'wet dog shake' behaviors in the guinea pig in a dose-dependent manner. Microdialysis studies demonstrated that acute administration of talnetant (30 mg/kg i.p.) produced significant increases in extracellular dopamine and norepinephrine in the medial prefrontal cortex and attenuated haloperidol-induced increases in nucleus accumbens dopamine levels in the freely moving guinea pigs. Taken together, these data demonstrate that talnetant is a selective, competitive, brain-penetrant NK3 receptor antagonist with the ability to modulate mesolimbic and mesocortical dopaminergic neurotransmission and hence support its potential therapeutic utility in the treatment of schizophrenia.
Subject(s)
Brain/drug effects , Quinolines/pharmacology , Receptors, Neurokinin-3/antagonists & inhibitors , Animals , Brain/cytology , Calcium/metabolism , Cell Line, Transformed , Cell Line, Tumor , Dopamine/metabolism , Dose-Response Relationship, Drug , Guinea Pigs , Humans , In Vitro Techniques , Male , Neurons/drug effects , Neurotransmitter Agents/metabolism , Protein Binding/drug effects , Receptors, Neurokinin-3/drug effects , Receptors, Neurokinin-3/metabolismABSTRACT
6-[(3-Cyclobutyl-2,3,4,5-tetrahydro-1H-3-benzazepin-7-yl)oxy]-N-methyl-3-pyridinecarboxamide hydrochloride (GSK189254) is a novel histamine H(3) receptor antagonist with high affinity for human (pK(i) = 9.59 -9.90) and rat (pK(i) = 8.51-9.17) H(3) receptors. GSK189254 is >10,000-fold selective for human H(3) receptors versus other targets tested, and it exhibited potent functional antagonism (pA(2) = 9.06 versus agonist-induced changes in cAMP) and inverse agonism [pIC(50) = 8.20 versus basal guanosine 5'-O-(3-[(35)S]thio)triphosphate binding] at the human recombinant H(3) receptor. In vitro autoradiography demonstrated specific [(3)H]GSK189254 binding in rat and human brain areas, including cortex and hippocampus. In addition, dense H(3) binding was detected in medial temporal cortex samples from severe cases of Alzheimer's disease, suggesting for the first time that H(3) receptors are preserved in late-stage disease. After oral administration, GSK189254 inhibited cortical ex vivo R-(-)-alpha-methyl[imidazole-2,5(n)-(3)H]histamine dihydrochloride ([(3)H]R-alpha-methylhistamine) binding (ED(50) = 0.17 mg/kg) and increased c-Fos immunoreactivity in prefrontal and somatosensory cortex (3 mg/kg). Microdialysis studies demonstrated that GSK189254 (0.3-3 mg/kg p.o.) increased the release of acetylcholine, noradrenaline, and dopamine in the anterior cingulate cortex and acetylcholine in the dorsal hippocampus. Functional antagonism of central H(3) receptors was demonstrated by blockade of R-alpha-methylhistamine-induced dipsogenia in rats (ID(50) = 0.03 mg/kg p.o.). GSK189254 significantly improved performance of rats in diverse cognition paradigms, including passive avoidance (1 and 3 mg/kg p.o.), water maze (1 and 3 mg/kg p.o.), object recognition (0.3 and 1 mg/kg p.o.), and attentional set shift (1 mg/kg p.o.). These data suggest that GSK189254 may have therapeutic potential for the symptomatic treatment of dementia in Alzheimer's disease and other cognitive disorders.
Subject(s)
Benzazepines/pharmacology , Brain/drug effects , Histamine Antagonists/pharmacology , Niacinamide/analogs & derivatives , Nootropic Agents/pharmacology , Receptors, Histamine H3/metabolism , Adolescent , Adult , Aged , Aged, 80 and over , Alzheimer Disease/metabolism , Alzheimer Disease/pathology , Alzheimer Disease/physiopathology , Animals , Benzazepines/metabolism , Benzazepines/pharmacokinetics , Binding, Competitive , Brain/metabolism , Brain/pathology , Cell Line , Dogs , Histamine Agonists/metabolism , Histamine Agonists/pharmacokinetics , Histamine Agonists/pharmacology , Histamine Antagonists/metabolism , Histamine Antagonists/pharmacokinetics , Humans , Male , Maze Learning/drug effects , Mice , Middle Aged , Neurotransmitter Agents/metabolism , Niacinamide/metabolism , Niacinamide/pharmacokinetics , Niacinamide/pharmacology , Nootropic Agents/metabolism , Nootropic Agents/pharmacokinetics , Rats , Rats, Sprague-Dawley , Rats, Wistar , Receptors, Histamine H3/analysis , Sus scrofaABSTRACT
SB-277011-A is a dopamine D(3) receptor antagonist that exhibits over 100-fold selectivity over dopamine D(2) receptors and a broad spectrum of other receptor, ion channels, and enzymes. We employed c-Fos immunohistochemistry to characterise the functional neuroanatomical effects of acute administration of SB-277011-A and observed a time-dependent increase in the density of c-Fos-like positive nuclei in rat forebrain with maximal effects observed 2 h post-dose. The relative influence of the different brain regions on the overall effect of SB-277011-A was ranked by partial least squares discriminant analysis loadings plot which indicated that sites within the nucleus accumbens exerted the greatest influence on the separation of the vehicle and SB-277011-A treatment groups. At the 2 h time-point, c-Fos-like expression was shown to be significantly elevated (p<0.05) in the core and shell of the nucleus accumbens, at both rostral and caudal levels, and in the lateral septum. No significant changes were detected in the caudate nucleus (lateral or medial) or in the cingulate, infralimbic prefrontal, or somatosensory cortices. The capacity of SB-277011-A to trigger immediate early gene expression in these limbic regions of rat brain adds to a growing consensus of the potential utility of dopamine D(3) receptor antagonism in psychiatric disorders including schizophrenia and drug dependency.
Subject(s)
Nitriles/pharmacology , Prosencephalon/drug effects , Proto-Oncogene Proteins c-fos/drug effects , Receptors, Dopamine D3/antagonists & inhibitors , Tetrahydroisoquinolines/pharmacology , Animals , Immunohistochemistry , Male , Prosencephalon/metabolism , Proto-Oncogene Proteins c-fos/biosynthesis , Rats , Rats, Sprague-Dawley , Time FactorsABSTRACT
Recent clinical evidence indicates that the broad spectrum anticonvulsant drug lamotrigine is effective against the depressive phase of bipolar illness and the difficult to treat rapid cycling form of the disorder. However, the molecular mechanism underlying this therapeutic action remains uncertain. Given that inhibition of the A-type of monoamine oxidase (MAO) is a proven antidepressant mechanism, we investigated the effects of lamotrigine on MAO activities in vitro and on monoamine disposition in vivo. In vitro, lamotrigine inhibited rat brain MAO activities with Ki values (MAO-A, 15 microM; MAO-B, 18 microM) potentially within the therapeutic range for this drug. The effects of lamotrigine on the MAO-A activities of rat brain and human liver preparations were almost identical suggesting minimal species or tissue variation. In contrast, there was no (MAO-A) or minimal (MAO-B) reduction in brain MAO activities when assayed ex vivo following the administration of lamotrigine to rats. In vivo brain microdialysis failed to detect meaningful alterations in extracellular hippocampal or frontal cortex monoamine concentrations. Furthermore, lamotrigine did not modulate oral tyramine-induced hypertension in rats or 5-hydroxytryptophan-induced head shaking in mice, providing strong evidence that the drug does not perturb monoamine metabolism in vivo. The absence of observable effects of lamotrigine on monoamine disposition in vivo may be explained by the competitive and highly reversible nature of the interaction of lamotrigine with MAO isoforms. Thus, altered monoamine metabolism in vivo is unlikely to account for the antidepressant action of the drug in bipolar depression.
Subject(s)
Anticonvulsants/pharmacology , Biogenic Monoamines/metabolism , Monoamine Oxidase/metabolism , Triazines/pharmacology , 5-Hydroxytryptophan/pharmacology , Administration, Oral , Animals , Behavior, Animal/drug effects , Blood Pressure/drug effects , Brain/drug effects , Brain/enzymology , Cerebellum/drug effects , Cerebellum/enzymology , Dopamine/metabolism , Dose-Response Relationship, Drug , Extracellular Space/drug effects , Extracellular Space/metabolism , Humans , Lamotrigine , Liver/drug effects , Liver/enzymology , Male , Microdialysis , Norepinephrine/metabolism , Rats , Rats, Wistar , Serotonin/metabolism , Time Factors , Tyramine/pharmacologyABSTRACT
RATIONALE: The alpha7 subtype of the nicotinic receptor plays an important role in auditory sensory gating. Schizophrenics show deficient sensory gating and abnormalities in the number and regulation of nicotinic receptors. Prepulse inhibition (PPI) deficits exhibited by isolation-reared rats are thought to model the sensorimotor gating deficits seen in schizophrenia. OBJECTIVE: To examine the role of nicotinic alpha7 receptors in the isolation-rearing rat model, we tested whether the selective alpha7 receptor agonist (R)-N-(1-Azabicyclo[2.2.2]oct-3-yl)(5-(2-pyridyl)thiophene-2-carboxamide) (compound A) could reverse isolation-rearing-induced PPI deficits, and investigated alpha7 receptor RNA expression in the hippocampus, prefrontal cortex, cerebellum, nucleus accumbens and thalamus, and alpha7 receptor protein expression in the hippocampus of isolation- and group-reared rats. METHOD: Rats reared in isolation or groups of five from weaning were tested in the PPI paradigm under conditions of variable inter-stimulus interval (ISI) (pulse = 110 dB/50 ms; prepulse = 75 dB/30 ms; ISI = 30, 100 and 300 ms) 30 min following administration of compound A (3.2-10 mg/kg i.p.). Alpha7 receptor expression was measured by TaqMan RT-PCR (total RNA) and autoradiography (protein). RESULTS: Isolation-rearing-induced PPI deficits were attenuated by both doses of compound A at 100-ms ISI and by 10 mg/kg at 300-ms ISI. Expression of alpha7 receptor RNA and protein was unaltered in isolation-reared rats. CONCLUSION: Although altered alpha7 receptor expression may not underlie the phenotype of isolation-reared rats, the activation of these receptors may be of benefit in re-establishing efficient gating function.
Subject(s)
Nicotinic Agonists/pharmacology , Receptors, Nicotinic/physiology , Reflex, Startle/drug effects , Social Isolation , Aconitine/analogs & derivatives , Aconitine/metabolism , Animals , Bungarotoxins/metabolism , Male , RNA, Messenger/analysis , Rats , Receptors, Nicotinic/genetics , Reverse Transcriptase Polymerase Chain Reaction , alpha7 Nicotinic Acetylcholine ReceptorABSTRACT
Chandelier neurons and their characteristic arrays of axonal terminals, known as cartridges, have been implicated in a variety of psychiatric and neurological disorders including schizophrenia and epilepsy. As a result, these neurons have been extensively examined in the brains of several species using a range of markers. However, these markers have not been systematically compared in a single species for their robustness in labelling chandelier cell cartridges. We have therefore examined several markers, reported to label chandelier arrays in primates, for their capacity to mark these structures in rat medial prefrontal cortex and hippocampus. These studies revealed that cartridge-like structures were labelled by parvalbumin and GAT-1 immunohistochemistry in both medial prefrontal cortex and hippocampus of the rat brain. Additionally, GAD65 immunohistochemistry labelled array-like structures preferentially in the dentate gyrus. In contrast, PSA-NCAM, calbindin and GAD67 immunohistochemistry did not reveal any array-like structures in either region of rat brain. These observations indicate that the various immunological markers previously used to visualise chandelier cell cartridges in primates are not equally efficient in labelling these structures in the rat brain, and that GAT-1 immunohistochemistry is the most robust means of visualising chandelier cell cartridges in the regions examined. These are important considerations for quantitative studies in animal models of neurological disorders where chandelier neurons are implicated.
Subject(s)
Hippocampus/cytology , Interneurons/metabolism , Membrane Transport Proteins/metabolism , Parvalbumins/metabolism , Prefrontal Cortex/cytology , Animals , Biomarkers/metabolism , Calcium-Binding Proteins/metabolism , GABA Plasma Membrane Transport Proteins , Hippocampus/metabolism , Immunohistochemistry , Interneurons/classification , Interneurons/cytology , Male , Nerve Tissue Proteins/metabolism , Prefrontal Cortex/metabolism , Presynaptic Terminals/classification , Presynaptic Terminals/metabolism , Rats , Rats, Inbred Strains , gamma-Aminobutyric Acid/metabolismABSTRACT
Epilepsy is perhaps the most common of all the serious neurological disorders, with 50-100 million people worldwide exhibiting clinically recognized epilepsy. However, there has been relatively little improvement in anticonvulsant drug efficacy since the introduction of, for example, phenobarbital and phenytoin in 1912 and 1930. Epilepsy can be broadly subdivided into partial, generalized and unclassified seizures, and seizure type is used to guide the selection of the anticonvulsant drug to be used. If monotherapy with one anticonvulsant class fails, then an agent from a different class is attempted. If patients still prove to be refractory, combination therapy is considered. Despite side effects, approximately 60% of patients experience long-term remission of their symptoms following drug therapy, while 40% remain refractory to drug treatment, emphasizing the need to develop novel anticonvulsant mechanisms with enhanced efficacy.
Subject(s)
Anticonvulsants/therapeutic use , Epilepsy/drug therapy , Animals , Calcium Channel Blockers/therapeutic use , Humans , Receptors, AMPA/physiology , Receptors, GABA-A/physiology , Receptors, N-Methyl-D-Aspartate/physiology , Sodium Channel Blockers/therapeutic useABSTRACT
The novel putative anticonvulsant drug 1-[2,6-difluorophenyl)-methyl]-1H-1,2,3-triazolo[4,5-c]) pyridine-4-amine monohydrochloride (BW534U87) effectively reduced seizures induced in rodents by threshold maximal and supramaximal electroshock, electrical kindling, pentylenetetrazole (PTZ) infusion and by vestibular stimulation in the genetically seizure-prone epilepsy-like (EL) mouse. The range of animal seizure models in which BW534U87 was effective is consistent with a broad spectrum anticonvulsant profile. In the EL mouse, the activity of BW534U87 was partially reversed by predosing with the selective adenosine A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), suggesting that an adenosine-dependent mechanism contributed to the antiseizure activity of the drug. BW534U87 inhibited rat brain homogenate adenosine deaminase activity, thus, raising the possibility that, by blocking the metabolism of endogenous adenosine by this route, BW534U87 limited seizure activity by promoting the inhibitory tone mediated by endogenous adenosine in the brain. The seizure protection conferred by the selective adenosine deaminase inhibitor erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) in EL mice and mice infused with PTZ confirms that inhibition of adenosine metabolism by deamination is an effective antiseizure strategy in these models.
Subject(s)
Adenine/analogs & derivatives , Adenosine/physiology , Anticonvulsants/pharmacology , Triazoles/pharmacology , Adenosine Deaminase/metabolism , Adenosine Deaminase Inhibitors , Amygdala/physiology , Animals , Convulsants , Dose-Response Relationship, Drug , Electroshock , Enzyme Inhibitors/pharmacology , Female , Kindling, Neurologic/drug effects , Male , Mice , Pentylenetetrazole , Purinergic P1 Receptor Antagonists , Rats , Rats, Wistar , Seizures/chemically induced , Seizures/prevention & control , Xanthines/pharmacologyABSTRACT
Exposure of adult rat cerebellar slices to a moderately raised K+ concentration (15 mM) caused a large (30-fold) rise in the levels of cyclic GMP. Excitatory amino acid antagonists failed to inhibit this response, nor could it be mimicked by agonists active at a number of other transmitter receptors. It was, however, inhibited by the nitric oxide (NO) synthase antagonist, l-methylarginine (IC50=10 microM), and also by tetrodotoxin (1 microM) implying that underlying the cyclic GMP response was an action potential-dependent formation of NO. Prelesioning of climbing fibres resulted in a loss of approximately 50% of the response to K+ but failed to influence the effects of glutamate receptor agonists or the NO-donor, nitroprusside. These findings point to a new mechanism for the formation of NO in the central nervous system and suggest that, in the cerebellum, climbing fibres are a source of NO.