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1.
Cereb Cortex ; 29(6): 2424-2436, 2019 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-29771287

RESUMO

Epidemiological studies have provided contradictory data on the deleterious sequels of cesarean section (C-section) delivery and their links with developmental brain disorders such as Autism Spectrum Disorders. To gain better insight on these issues, we have now compared physiological, morphological, and behavioral parameters in vaginal, term, and preterm C-section delivered mice. We report that C-section delivery does not lead to long-term behavioral alterations though preterm C-section delivery modifies communicative behaviors in pups. Moreover, C-section delivery neither alters the gamma-aminobutyric acid (GABA) developmental excitatory to inhibitory shift nor the frequency or amplitude of glutamatergic and GABAergic postsynaptic currents in hippocampal pyramidal neurons. However, these neurons present an underdeveloped dendritic arbor at birth in pups born by C-section delivery, but this difference disappears 1 day later suggesting an accelerated growth after birth. Therefore, C-section delivery, with prematurity as an aggravating factor, induces transient developmental delays but neither impacts the GABA developmental sequence nor leads to long-term consequences in mice. The deleterious sequels of C-section delivery described in epidemiological studies might be due to a perinatal insult that could be aggravated by C-section delivery.


Assuntos
Cesárea/efeitos adversos , Deficiências do Desenvolvimento/epidemiologia , Nascimento Prematuro , Animais , Comportamento Animal/fisiologia , Região CA3 Hipocampal/metabolismo , Região CA3 Hipocampal/patologia , Região CA3 Hipocampal/fisiopatologia , Feminino , Masculino , Camundongos , Gravidez , Células Piramidais/metabolismo , Células Piramidais/patologia
2.
Cereb Cortex ; 29(9): 3982-3992, 2019 08 14.
Artigo em Inglês | MEDLINE | ID: mdl-30395185

RESUMO

Epidemiological and experimental studies suggest that maternal immune activation (MIA) leads to developmental brain disorders, but whether the pathogenic mechanism impacts neurons already at birth is not known. We now report that MIA abolishes in mice the oxytocin-mediated delivery γ-aminobutyric acid (GABA) shift from depolarizing to hyperpolarizing in CA3 pyramidal neurons, and this is restored by the NKCC1 chloride importer antagonist bumetanide. Furthermore, MIA hippocampal pyramidal neurons at birth have a more exuberant apical arbor organization and increased apical dendritic length than age-matched controls. The frequency of spontaneous glutamatergic postsynaptic currents is also increased in MIA offspring, as well as the pairwise correlation of the synchronized firing of active cells in CA3. These alterations produced by MIA persist, since at P14-15 GABA action remains depolarizing, produces excitatory action, and network activity remains elevated with a higher frequency of spontaneous glutamatergic postsynaptic currents. Therefore, the pathogenic actions of MIA lead to important morphophysiological and network alterations in the hippocampus already at birth.


Assuntos
Região CA3 Hipocampal/crescimento & desenvolvimento , Região CA3 Hipocampal/imunologia , Potenciais da Membrana , Gravidez/imunologia , Células Piramidais/imunologia , Ácido gama-Aminobutírico/imunologia , Animais , Região CA3 Hipocampal/efeitos dos fármacos , Dendritos/efeitos dos fármacos , Dendritos/imunologia , Feminino , Ácido Glutâmico/fisiologia , Potenciais da Membrana/efeitos dos fármacos , Camundongos Endogâmicos C57BL , Poli I-C/administração & dosagem , Células Piramidais/citologia , Células Piramidais/efeitos dos fármacos , Membro 2 da Família 12 de Carreador de Soluto/imunologia
3.
Cereb Cortex ; 29(9): 3778-3795, 2019 08 14.
Artigo em Inglês | MEDLINE | ID: mdl-30295710

RESUMO

Epilepsy is a multifactorial disorder associated with neuronal hyperexcitability that affects more than 1% of the human population. It has long been known that adenosine can reduce seizure generation in animal models of epilepsies. However, in addition to various side effects, the instability of adenosine has precluded its use as an anticonvulsant treatment. Here we report that a stable analogue of diadenosine-tetraphosphate: AppCH2ppA effectively suppresses spontaneous epileptiform activity in vitro and in vivo in a Tuberous Sclerosis Complex (TSC) mouse model (Tsc1+/-), and in postsurgery cortical samples from TSC human patients. These effects are mediated by enhanced adenosine signaling in the cortex post local neuronal adenosine release. The released adenosine induces A1 receptor-dependent activation of potassium channels thereby reducing neuronal excitability, temporal summation, and hypersynchronicity. AppCH2ppA does not cause any disturbances of the main vital autonomous functions of Tsc1+/- mice in vivo. Therefore, we propose this compound to be a potent new candidate for adenosine-related treatment strategies to suppress intractable epilepsies.


Assuntos
Adenosina/fisiologia , Anticonvulsivantes/administração & dosagem , Fosfatos de Dinucleosídeos/administração & dosagem , Neocórtex/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Convulsões/fisiopatologia , Animais , Feminino , Humanos , Masculino , Potenciais da Membrana/efeitos dos fármacos , Camundongos , Camundongos Transgênicos , Neocórtex/fisiopatologia , Neurônios/fisiologia , Canais de Potássio/fisiologia , Receptor A1 de Adenosina/fisiologia , Convulsões/prevenção & controle , Transdução de Sinais/efeitos dos fármacos , Proteína 1 do Complexo Esclerose Tuberosa/genética
4.
Neural Plast ; 2019: 2382639, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31354805

RESUMO

Autism spectrum disorders (ASD) are neurodevelopmental disorders induced by genetic and environmental factors. In our recent studies, we showed that the GABA developmental shifts during delivery and the second postnatal week are abolished in two rodent models of ASD. Maternal treatment around birth with bumetanide restored the GABA developmental sequence and attenuated the autism pathogenesis in offspring. Clinical trials conducted in parallel confirmed the usefulness of bumetanide treatment to attenuate the symptoms in children with ASD. Collectively, these observations suggest that an alteration of the GABA developmental sequence is a hallmark of ASD. Here, we investigated whether similar alterations occur in the Shank3 mouse model of ASD. We report that in CA3 pyramidal neurons, the driving force and inhibitory action of GABA are not different in naïve and Shank3-mutant age-matched animals at birth and during the second postnatal week. In contrast, the frequency of spontaneous excitatory postsynaptic currents is already enhanced at birth and persists through postnatal day 15. Therefore, in CA3 pyramidal neurons of Shank3-mutant mice, glutamatergic but not GABAergic activity is affected at early developmental stages, hence reflecting the heterogeneity of mechanisms underlying the pathogenesis of ASD.


Assuntos
Região CA3 Hipocampal/metabolismo , Potenciais Pós-Sinápticos Excitadores/fisiologia , Ácido Glutâmico/metabolismo , Proteínas do Tecido Nervoso/genética , Células Piramidais/metabolismo , Animais , Animais Recém-Nascidos , Transtorno do Espectro Autista/genética , Transtorno do Espectro Autista/metabolismo , Modelos Animais de Doenças , Camundongos , Camundongos Knockout , Proteínas dos Microfilamentos , Proteínas do Tecido Nervoso/metabolismo , Técnicas de Patch-Clamp , Ácido gama-Aminobutírico/metabolismo
5.
Mol Pain ; 122016.
Artigo em Inglês | MEDLINE | ID: mdl-27030723

RESUMO

BACKGROUND: A growing body of evidence suggests that ATP-gated P2X3 receptors (P2X3Rs) are implicated in chronic pain. We address the possibility that stable, synthetic analogs of diadenosine tetraphosphate (Ap4A) might induce antinociceptive effects by inhibiting P2X3Rs in peripheral sensory neurons. RESULTS: The effects of two stable, synthetic Ap4A analogs (AppNHppA and AppCH2ppA) are studied firstly in vitro on HEK293 cells expressing recombinant rat P2XRs (P2X2Rs, P2X3Rs, P2X4Rs, and P2X7Rs) and then using native rat brain cells (cultured trigeminal, nodose, or dorsal root ganglion neurons). Thereafter, the action of these stable, synthetic Ap4A analogs on inflammatory pain and thermal hyperalgesia is studied through the measurement of antinociceptive effects in formalin and Hargreaves plantar tests in rats in vivo. In vitro inhibition of rat P2X3Rs (not P2X2Rs, P2X4Rs nor P2X7Rs) is shown to take place mediated by high-affinity desensitization (at low concentrations; IC50 values 100-250 nM) giving way to only weak partial agonism at much higher concentrations (EC50 values ≥ 10 µM). Similar inhibitory activity is observed with human recombinant P2X3Rs. The inhibitory effects of AppNHppA on nodose, dorsal root, and trigeminal neuron whole cell currents suggest that stable, synthetic Ap4A analogs inhibit homomeric P2X3Rs in preference to heteromeric P2X2/3Rs. Both Ap4A analogs mediate clear inhibition of pain responses in both in vivo inflammation models. CONCLUSIONS: Stable, synthetic Ap4A analogs (AppNHppA and AppCH2ppA) being weak partial agonist provoke potent high-affinity desensitization-mediated inhibition of homomeric P2X3Rs at low concentrations. Therefore, both analogs demonstrate clear potential as potent analgesic agents for use in the management of chronic pain associated with heightened P2X3R activation.


Assuntos
Fosfatos de Dinucleosídeos/uso terapêutico , Inflamação/complicações , Inflamação/tratamento farmacológico , Dor/complicações , Dor/tratamento farmacológico , Antagonistas do Receptor Purinérgico P2X/uso terapêutico , Receptores Purinérgicos P2X3/metabolismo , Analgésicos/farmacologia , Analgésicos/uso terapêutico , Animais , Fosfatos de Dinucleosídeos/farmacologia , Células HEK293 , Humanos , Hiperalgesia/complicações , Hiperalgesia/tratamento farmacológico , Injeções Subcutâneas , Masculino , Multimerização Proteica/efeitos dos fármacos , Subunidades Proteicas/metabolismo , Antagonistas do Receptor Purinérgico P2X/farmacologia , Ratos Wistar , Proteínas Recombinantes/metabolismo , Células Receptoras Sensoriais/efeitos dos fármacos , Células Receptoras Sensoriais/metabolismo
6.
Cereb Cortex ; 25(9): 2440-55, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24646614

RESUMO

Cannabinoids are known to regulate inhibitory synaptic transmission via activation of presynaptic G protein-coupled cannabinoid CB1 receptors (CB1Rs). Additionally, recent studies suggest that cannabinoids can also directly interact with recombinant GABAA receptors (GABAARs), potentiating currents activated by micromolar concentrations of γ-aminobutyric acid (GABA). However, the impact of this direct interaction on GABAergic inhibition in central nervous system is unknown. Here we report that currents mediated by recombinant GABAARs activated by high (synaptic) concentrations of GABA as well as GABAergic inhibitory postsynaptic currents (IPSCs) at neocortical fast spiking (FS) interneuron to pyramidal neuron synapses are suppressed by exogenous and endogenous cannabinoids in a CB1R-independent manner. This IPSC suppression may account for disruption of inhibitory control of pyramidal neurons by FS interneurons. At FS interneuron to pyramidal neuron synapses, endocannabinoids induce synaptic low-pass filtering of GABAAR-mediated currents evoked by high-frequency stimulation. The CB1R-independent suppression of inhibition is synapse specific. It does not occur in CB1R containing hippocampal cholecystokinin-positive interneuron to pyramidal neuron synapses. Furthermore, in contrast to synaptic receptors, the activity of extrasynaptic GABAARs in neocortical pyramidal neurons is enhanced by cannabinoids in a CB1R-independent manner. Thus, cannabinoids directly interact differentially with synaptic and extrasynaptic GABAARs, providing a potent novel context-dependent mechanism for regulation of inhibition.


Assuntos
Canabinoides/metabolismo , Potenciais Pós-Sinápticos Inibidores/fisiologia , Inibição Neural/fisiologia , Receptores de GABA/metabolismo , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/genética , Animais , Animais Recém-Nascidos , Canabinoides/farmacologia , GABAérgicos/farmacologia , Hipocampo/citologia , Humanos , Técnicas In Vitro , Potenciais Pós-Sinápticos Inibidores/efeitos dos fármacos , Potenciais Pós-Sinápticos Inibidores/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Inibição Neural/efeitos dos fármacos , Técnicas de Patch-Clamp , Ratos , Ratos Sprague-Dawley , Ratos Wistar , Receptor CB1 de Canabinoide/genética , Receptor CB1 de Canabinoide/metabolismo , Transmissão Sináptica/efeitos dos fármacos , Transmissão Sináptica/genética , Transfecção
7.
Brain ; 136(Pt 8): 2457-73, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23831613

RESUMO

Altered development of the human cerebral cortex can cause severe malformations with often intractable focal epileptic seizures and may participate in common pathologies, notably epilepsy. This raises important conceptual and therapeutic issues. Two missense mutations in the sushi repeat-containing protein SRPX2 had been previously identified in epileptic disorders with or without structural developmental alteration of the speech cortex. In the present study, we aimed to decipher the precise developmental role of SRPX2, to have a better knowledge on the consequences of its mutations, and to start addressing therapeutic issues through the design of an appropriate animal model. Using an in utero Srpx2 silencing approach, we show that SRPX2 influences neuronal migration in the developing rat cerebral cortex. Wild-type, but not the mutant human SRPX2 proteins, rescued the neuronal migration phenotype caused by Srpx2 silencing in utero, and increased alpha-tubulin acetylation. Following in utero Srpx2 silencing, spontaneous epileptiform activity was recorded post-natally. The neuronal migration defects and the post-natal epileptic consequences were prevented early in embryos by maternal administration of tubulin deacetylase inhibitor tubacin. Hence epileptiform manifestations of developmental origin could be prevented in utero, using a transient and drug-based therapeutic protocol.


Assuntos
Anilidas/farmacologia , Movimento Celular/genética , Córtex Cerebral/metabolismo , Epilepsia/genética , Inibidores de Histona Desacetilases/farmacologia , Ácidos Hidroxâmicos/farmacologia , Proteínas de Membrana/genética , Neurônios/metabolismo , Animais , Movimento Celular/efeitos dos fármacos , Córtex Cerebral/citologia , Córtex Cerebral/efeitos dos fármacos , Epilepsia/metabolismo , Inativação Gênica , Humanos , Neurônios/citologia , Neurônios/efeitos dos fármacos , Ratos , Tubulina (Proteína)/genética , Tubulina (Proteína)/metabolismo
8.
eNeuro ; 11(5)2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38755010

RESUMO

Cholinergic neurons of the basal forebrain represent the main source of cholinergic innervation of large parts of the neocortex and are involved in adults in the modulation of attention, memory, and arousal. During the first postnatal days, they play a crucial role in the development of cortical neurons and cortical cytoarchitecture. However, their characteristics, during this period have not been studied. To understand how they can fulfill this role, we investigated the morphological and electrophysiological maturation of cholinergic neurons of the substantia innominata-nucleus basalis of Meynert (SI/NBM) complex in the perinatal period in mice. We show that cholinergic neurons, whether or not they express gamma-aminobutyric acid (GABA) as a cotransmitter, are already functional at Embryonic Day 18. Until the end of the first postnatal week, they constitute a single population of neurons with a well developed dendritic tree, a spontaneous activity including bursting periods, and a short-latency response to depolarizations (early-firing). They are excited by both their GABAergic and glutamatergic afferents. During the second postnatal week, a second, less excitable, neuronal population emerges, with a longer delay response to depolarizations (late-firing), together with the hyperpolarizing action of GABAA receptor-mediated currents. This classification into early-firing (40%) and late-firing (60%) neurons is again independent of the coexpression of GABAergic markers. These results strongly suggest that during the first postnatal week, the specific properties of developing SI/NBM cholinergic neurons allow them to spontaneously release acetylcholine (ACh), or ACh and GABA, into the developing cortex.


Assuntos
Prosencéfalo Basal , Neurônios Colinérgicos , Ácido gama-Aminobutírico , Animais , Neurônios Colinérgicos/fisiologia , Neurônios Colinérgicos/metabolismo , Ácido gama-Aminobutírico/metabolismo , Prosencéfalo Basal/fisiologia , Prosencéfalo Basal/metabolismo , Animais Recém-Nascidos , Camundongos Endogâmicos C57BL , Feminino , Núcleo Basal de Meynert/fisiologia , Núcleo Basal de Meynert/metabolismo , Substância Inominada/fisiologia , Substância Inominada/metabolismo , Camundongos , Receptores de GABA-A/metabolismo , Potenciais de Ação/fisiologia , Técnicas de Patch-Clamp , Ácido Glutâmico/metabolismo
9.
J Neurosci ; 32(50): 18047-53, 2012 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-23238720

RESUMO

Cellular electrophysiological signatures of Parkinson's disease described in the pharmacological 6-hydroxydopamine (6-OHDA) animal models of Parkinson's disease include spontaneous repetitive giant GABAergic currents in a subpopulation of striatal medium spiny neurons (MSNs), and spontaneous rhythmic bursts of spikes generated by subthalamic nucleus (STN) neurons. We investigated whether similar signatures are present in Pink1(-/-) mice, a genetic rodent model of the PARK6 variant of Parkinson's disease. Although 9- to 24-month-old Pink1(-/-) mice show reduced striatal dopamine content and release, and impaired spontaneous locomotion, the relevance of this model to Parkinson's disease has been questioned because mesencephalic dopaminergic neurons do not degenerate during the mouse lifespan. We show that 75% of the MSNs of 5- to 7-month-old Pink1(-/-) mice exhibit giant GABAergic currents, occurring either singly or in bursts (at 40 Hz), rather than the low-frequency (2 Hz), low-amplitude, tonic GABAergic drive common to wild-type MSNs of the same age. STN neurons from 5- to 7-month-old Pink1(-/-) mice spontaneously generated bursts of spikes instead of the control tonic drive. Chronic kainic acid lesion of the STN or chronic levodopa treatment reliably suppressed the giant GABAergic currents of MSNs after 1 month and replaced them with the control tonic activity. The similarity between the in vitro resting states of Pink1 MSNs and those of fully dopamine (DA)-depleted MSNs of 6-OHDA-treated mice, together with the beneficial effect of levodopa treatment, strongly suggest that dysfunction of mesencephalic dopaminergic neurons in Pink1(-/-) mice is more severe than expected. The beneficial effect of the STN lesion also suggests that pathological STN activity strongly influences striatal networks in Pink1(-/-) mice.


Assuntos
Levodopa/farmacologia , Neurônios/efeitos dos fármacos , Transtornos Parkinsonianos/fisiopatologia , Proteínas Quinases/deficiência , Núcleo Subtalâmico/efeitos dos fármacos , Ácido gama-Aminobutírico/metabolismo , Animais , Antiparkinsonianos/farmacologia , Corpo Estriado/patologia , Corpo Estriado/fisiopatologia , Condutividade Elétrica , Agonistas de Aminoácidos Excitatórios/toxicidade , Feminino , Imuno-Histoquímica , Ácido Caínico/toxicidade , Masculino , Camundongos , Camundongos Knockout , Neurônios/fisiologia , Transtornos Parkinsonianos/patologia , Técnicas de Patch-Clamp , Proteínas Quinases/genética , Núcleo Subtalâmico/lesões , Núcleo Subtalâmico/patologia , Núcleo Subtalâmico/fisiopatologia
10.
Commun Biol ; 6(1): 723, 2023 07 14.
Artigo em Inglês | MEDLINE | ID: mdl-37452171

RESUMO

Cholinergic interneurons of the striatum play a role in action selection and associative learning by activating local GABAergic inhibitory microcircuits. We investigated whether cholinergic-GABAergic microcircuits function differently and fulfill a different role during early postnatal development, when GABAA actions are not inhibitory and mice pups do not walk. We focused our study mainly on dual cholinergic/GABAergic interneurons (CGINs). We report that morphological and intrinsic electrophysiological properties of CGINs rapidly develop during the first post-natal week. At this stage, CGINs are excited by the activation of GABAA receptors or GABAergic synaptic inputs, respond to cortical stimulation by a long excitation and are linked by polysynaptic excitations. All these excitations are replaced by inhibitions at P12-P15. Early chronic treatment with the NKCC1 antagonist bumetanide to evoke premature GABAergic inhibitions from P4 to P8, prevented the GABA polarity shift and corticostriatal pause response at control postnatal days. We propose that early excitatory cholinergic-GABAergic microcircuits are instrumental in the maturation of GABAergic inhibition.


Assuntos
Colinérgicos , Potenciais Pós-Sinápticos Inibidores , Camundongos , Animais , Potenciais Pós-Sinápticos Inibidores/fisiologia , Colinérgicos/farmacologia , Corpo Estriado/metabolismo , Receptores de GABA-A/metabolismo , Ácido gama-Aminobutírico/farmacologia
11.
Cell Stress ; 2(6): 147-149, 2018 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-31225481

RESUMO

The rule of one terminal and one transmitter acting on one synapse clearly fails to cover the complexity of chemical synapse operation in the brain. Compelling evidence now indicates that two transmitters can be released from the same terminal, acting in a complementary manner to generate complex electrical activity in the targets. Our laboratory now showed that a subpopulation striatal cholinergic neurons also release the classical inhibitory transmitter GABA with a balance between excitation and inhibition being provided by acetylcholine and GABA, respectively. An illustration of the importance of this dual release comes from the fact that when dopamine signals are absent such as in Parkinson disease (PD) the GABAergic inhibition in these dual cholinergic/GABAergic cells fails because of high intracellular chloride ((Cl-)I) levels rendering the cholinergic excitatory component unmet by a parallel inhibitory drive. Restoring low (Cl-)I with the NKCC1 chloride importer antagonist bumetanide attenuates the electrical and motor disturbance. In addition to illustrating the complex interactions between two transmitters acting at the same synapse, this study paves the way to novel conceptual treatment of PD based on restoration of GABAergic inhibition in keeping with our pilot clinical trial showing indeed that bumetanide together with levodopa attenuates axial motor disturbance. It is also in keeping with extensive investigations showing increased (Cl-)I levels and weakened inhibition in a wide range of pathological insults and their restoration by bumetanide. It raises fundamental issues related to the operation of the striatum and basal ganglia in health and disease.

12.
Methods Mol Biol ; 1677: 231-239, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28986876

RESUMO

Analysis of electrophysiological properties of NMDARs and NMDAR-mediated synaptic transmission in identified neurons and synapses in brain slices is a major step in understanding their function in normal and pathological neuronal brain networks. In many central synapses excitatory postsynaptic currents (EPSCs) are mediated by excitatory neurotransmitter glutamate that activates colocalized AMPAR and NMDAR generating a complex EPSC. Here, we describe the methods commonly used in brain slices to study the electrophysiological properties of NMDAR-mediated component of spontaneous or evoked EPSCs by extracellular stimulation or by stimulating synaptically connected neurons. This approach is based on whole-cell patch-clamp recordings, pharmacological tools, and the analysis of the difference in temporal parameters between the AMPA and NMDA receptors. It allows pinpointing of the basic functional properties of NMDARs that are specific to identified brain regions, neurons, and synapses of wild-type or genetically manipulated mice.


Assuntos
Encéfalo/metabolismo , Receptores de AMPA/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Animais , Potenciais Pós-Sinápticos Excitadores/fisiologia , Neurônios/metabolismo , Técnicas de Patch-Clamp , Sinapses/metabolismo
13.
J Neurosci ; 25(33): 7499-506, 2005 Aug 17.
Artigo em Inglês | MEDLINE | ID: mdl-16107637

RESUMO

At many central synapses, endocannabinoids released by postsynaptic cells act retrogradely on presynaptic G-protein-coupled cannabinoid receptors to inhibit neurotransmitter release. Here, we demonstrate that cannabinoids may directly affect the functioning of inhibitory glycine receptor (GlyR) channels. In isolated hippocampal pyramidal and Purkinje cerebellar neurons, endogenous cannabinoids anandamide and 2-arachidonylglycerol, applied at physiological concentrations, inhibited the amplitude and altered the kinetics of rise time, desensitization, and deactivation of the glycine-activated current (I(Gly)) in a concentration-dependent manner. These effects of cannabinoids were observed in the presence of cannabinoid CB1/CB3, vanilloid receptor 1 antagonists, and the G-protein inhibitor GDPbetaS, suggesting a direct action of cannabinoids on GlyRs. The effect of cannabinoids on I(Gly) desensitization was strongly voltage dependent. We also demonstrate that, in the presence of a GABA(A) receptor antagonist, GlyRs may contribute to the generation of seizure-like activity induced by short bursts (seven stimuli) of high-frequency stimulation of inputs to hippocampal CA1 region, because this activity was diminished by selective GlyR antagonists (strychnine and ginkgolides B and J). The GlyR-mediated rhythmic activity was also reduced by cannabinoids (anandamide) in the presence of a CB1 receptor antagonist. These results suggest that the direct inhibition of GlyRs by endocannabinoids can modulate the hippocampal network activity.


Assuntos
Moduladores de Receptores de Canabinoides/fisiologia , Inibição Neural/fisiologia , Neurônios/efeitos dos fármacos , Neurônios/fisiologia , Receptores de Glicina/agonistas , Receptores de Glicina/fisiologia , Animais , Agonistas de Receptores de Canabinoides , Sistema Nervoso Central/efeitos dos fármacos , Sistema Nervoso Central/fisiologia , Relação Dose-Resposta a Droga , Técnicas In Vitro , Ratos , Ratos Wistar , Receptores de Canabinoides/fisiologia
14.
Biochim Biophys Acta ; 1745(1): 1-6, 2005 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-16085050

RESUMO

Acid-sensing ion channels (ASICs) are widely expressed in mammalian sensory neurons and supposedly play a role in nociception and acid sensing. In the course of functioning the redox status of the tissue is subjected to changes. Using whole-cell patch-clamp/concentration clamp techniques we have investigated the effect of redox reagents on the ASIC-like currents in the sensory ganglia and hippocampal neurons of rat. The reducing agent dithiothreitol (DTT), when applied in the concentrations 1-2 mM, reversibly potentiates proton-activated currents, while the oxidizing reagent 5,5'-dithio-bis-(2-nitrobenzoic acid) (DTNB) causes their inhibition. The EC50 and Hill coefficient for the activation of ASIC-like currents by protons are not affected by DTT. Redox modulation of proton-activated currents is independent on the membrane potential and on the level of pH used for the current activation. The endogenous antioxidant tripeptide glutathione (its reduced form, g-l-glutamyl-l-cysteinyl-glycine, GSH) also potentiates proton-activated currents. Our results indicate that ASIC-like currents are susceptible to regulation by redox agents.


Assuntos
Hipocampo/fisiologia , Concentração de Íons de Hidrogênio , Proteínas de Membrana/fisiologia , Proteínas do Tecido Nervoso/fisiologia , Células Piramidais/fisiologia , Canais de Sódio/fisiologia , Canais Iônicos Sensíveis a Ácido , Animais , Cálcio/metabolismo , Ácido Ditionitrobenzoico/farmacologia , Glutationa/farmacologia , Indicadores e Reagentes , Proteínas de Membrana/efeitos dos fármacos , Proteínas do Tecido Nervoso/efeitos dos fármacos , Oxirredução , Ratos , Ratos Wistar , Sódio/metabolismo , Canais de Sódio/efeitos dos fármacos
15.
Toxicology ; 207(1): 129-36, 2005 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-15590128

RESUMO

We have identified a novel polypeptide toxin (Lsp-1) from the venom of the spider Lycosa (LS). Its effect has been examined on the P-type calcium channels in Purkinje neurons, using whole-cell patch-clamp. This toxin (at saturating concentration 7 nM) produces prominent (four-fold) deceleration of the activation kinetics and partial (71+/-6%) decrease of the amplitude of P-current without affecting either deactivation or inactivation kinetics. These effects are not use-dependent. They are partially reversible within a minute upon the wash-out of the toxin. Intracellular perfusion of Purkinje neurons with 100 microM of GDP or 2 microM of GTPgammaS, as well as strong depolarising pre-pulses (+100 mV), do not eliminate the action of Lsp-1 on P-channels indicating that down-modulation via guanine nucleotide-binding proteins (G-proteins) is not involved in the observed phenomenon. In view of extremely high functional significance of P-channels, the toxin can be suggested as a useful pharmacological tool.


Assuntos
Canais de Cálcio Tipo P/metabolismo , Peptídeos/farmacologia , Células de Purkinje/efeitos dos fármacos , Venenos de Aranha/química , Toxinas Biológicas/farmacologia , Animais , Cinética , Potenciais da Membrana/efeitos dos fármacos , Técnicas de Patch-Clamp , Células de Purkinje/metabolismo , Células de Purkinje/fisiologia , Ratos , Ratos Wistar
16.
Science ; 343(6171): 675-9, 2014 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-24503856

RESUMO

We report that the oxytocin-mediated neuroprotective γ-aminobutyric acid (GABA) excitatory-inhibitory shift during delivery is abolished in the valproate and fragile X rodent models of autism. During delivery and subsequently, hippocampal neurons in these models have elevated intracellular chloride levels, increased excitatory GABA, enhanced glutamatergic activity, and elevated gamma oscillations. Maternal pretreatment with bumetanide restored in offspring control electrophysiological and behavioral phenotypes. Conversely, blocking oxytocin signaling in naïve mothers produced offspring having electrophysiological and behavioral autistic-like features. Our results suggest a chronic deficient chloride regulation in these rodent models of autism and stress the importance of oxytocin-mediated GABAergic inhibition during the delivery process. Our data validate the amelioration observed with bumetanide and oxytocin and point to common pathways in a drug-induced and a genetic rodent model of autism.


Assuntos
Transtorno Autístico/induzido quimicamente , Transtorno Autístico/genética , Citoproteção , Ocitocina/metabolismo , Ácido gama-Aminobutírico/metabolismo , Animais , Transtorno Autístico/metabolismo , Comportamento Animal , Bumetanida/administração & dosagem , Cloretos/metabolismo , Modelos Animais de Doenças , Feminino , Proteína do X Frágil da Deficiência Intelectual/genética , Troca Materno-Fetal , Camundongos , Parto , Gravidez , Ratos , Ácido Valproico/farmacologia
17.
Nat Genet ; 45(9): 1073-6, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23933818

RESUMO

Epilepsy-aphasia syndromes (EAS) are a group of rare, severe epileptic encephalopathies of unknown etiology with a characteristic electroencephalogram (EEG) pattern and developmental regression particularly affecting language. Rare pathogenic deletions that include GRIN2A have been implicated in neurodevelopmental disorders. We sought to delineate the pathogenic role of GRIN2A in 519 probands with epileptic encephalopathies with diverse epilepsy syndromes. We identified four probands with GRIN2A variants that segregated with the disorder in their families. Notably, all four families presented with EAS, accounting for 9% of epilepsy-aphasia cases. We did not detect pathogenic variants in GRIN2A in other epileptic encephalopathies (n = 475) nor in probands with benign childhood epilepsy with centrotemporal spikes (n = 81). We report the first monogenic cause, to our knowledge, for EAS. GRIN2A mutations are restricted to this group of cases, which has important ramifications for diagnostic testing and treatment and provides new insights into the pathogenesis of this debilitating group of conditions.


Assuntos
Síndrome de Landau-Kleffner/genética , Mutação , Receptores de N-Metil-D-Aspartato/genética , Eletroencefalografia , Feminino , Humanos , Síndrome de Landau-Kleffner/diagnóstico , Masculino , Linhagem , Fenótipo
18.
Nat Genet ; 45(9): 1061-6, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23933820

RESUMO

Epileptic encephalopathies are severe brain disorders with the epileptic component contributing to the worsening of cognitive and behavioral manifestations. Acquired epileptic aphasia (Landau-Kleffner syndrome, LKS) and continuous spike and waves during slow-wave sleep syndrome (CSWSS) represent rare and closely related childhood focal epileptic encephalopathies of unknown etiology. They show electroclinical overlap with rolandic epilepsy (the most frequent childhood focal epilepsy) and can be viewed as different clinical expressions of a single pathological entity situated at the crossroads of epileptic, speech, language, cognitive and behavioral disorders. Here we demonstrate that about 20% of cases of LKS, CSWSS and electroclinically atypical rolandic epilepsy often associated with speech impairment can have a genetic origin sustained by de novo or inherited mutations in the GRIN2A gene (encoding the N-methyl-D-aspartate (NMDA) glutamate receptor α2 subunit, GluN2A). The identification of GRIN2A as a major gene for these epileptic encephalopathies provides crucial insights into the underlying pathophysiology.


Assuntos
Epilepsias Parciais/genética , Síndrome de Landau-Kleffner/genética , Mutação , Receptores de N-Metil-D-Aspartato/genética , Substituição de Aminoácidos , Linhagem Celular , Eletroencefalografia , Feminino , Expressão Gênica , Genótipo , Humanos , Masculino , Linhagem , Fenótipo
19.
Front Mol Neurosci ; 4: 13, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21847369

RESUMO

Endocannabinoids are known as retrograde messengers, being released from the postsynaptic neuron and acting on specific presynaptic G-protein-coupled cannabinoid (CB) receptors to decrease neurotransmitter release. Also, at physiologically relevant concentrations cannabinoids can directly modulate the function of voltage-gated and receptor-operated ion channels. Using patch-clamp recording we analyzed the consequences of the direct action of an endocannabinoid, 2-arachidonoylglycerol (2-AG), on the functional properties of glycine receptor channels (GlyRs) and ionic currents in glycinergic synapses. At physiologically relevant concentrations (0.1-1 µM), 2-AG directly affected the functions of recombinant homomeric α1H GlyR: it inhibited peak amplitude and dramatically enhanced desensitization. The action of 2-AG on GlyR-mediated currents developed rapidly, within ∼300 ms. Addition of 1 µM 2-AG strongly facilitated the depression of glycine-induced currents during repetitive (4-10 Hz) application of short (2 ms duration) pulses of glycine to outside-out patches. In brainstem slices from CB1 receptor knockout mice, 2-AG significantly decreased the extent of facilitation of synaptic currents in hypoglossal motoneurons during repetitive (10-20 Hz) stimulation. These observations suggest that endocannabinoids can modulate postsynaptic metaplasticity of glycinergic synaptic currents in a CB1 receptor-independent manner.

20.
Science ; 327(5973): 1614-8, 2010 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-20150444

RESUMO

Synaptic vesicle fusion in brain synapses occurs in phases that are either tightly coupled to action potentials (synchronous), immediately following action potentials (asynchronous), or as stochastic events in the absence of action potentials (spontaneous). Synaptotagmin-1, -2, and -9 are vesicle-associated Ca2+ sensors for synchronous release. Here we found that double C2 domain (Doc2) proteins act as Ca2+ sensors to trigger spontaneous release. Although Doc2 proteins are cytosolic, they function analogously to synaptotagmin-1 but with a higher Ca2+ sensitivity. Doc2 proteins bound to N-ethylmaleimide-sensitive factor attachment receptor (SNARE) complexes in competition with synaptotagmin-1. Thus, different classes of multiple C2 domain-containing molecules trigger synchronous versus spontaneous fusion, which suggests a general mechanism for synaptic vesicle fusion triggered by the combined actions of SNAREs and multiple C2 domain-containing proteins.


Assuntos
Proteínas de Ligação ao Cálcio/metabolismo , Cálcio/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Neurotransmissores/metabolismo , Transmissão Sináptica , Vesículas Sinápticas/fisiologia , Potenciais de Ação , Animais , Sítios de Ligação , Proteínas de Ligação ao Cálcio/química , Proteínas de Ligação ao Cálcio/genética , Células Cultivadas , Potenciais Pós-Sinápticos Excitadores , Hipocampo/citologia , Potenciais Pós-Sinápticos Inibidores , Fusão de Membrana , Camundongos , Camundongos Knockout , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Proteínas do Tecido Nervoso/química , Proteínas do Tecido Nervoso/genética , Neurônios/fisiologia , Técnicas de Patch-Clamp , Estrutura Terciária de Proteína , Células de Purkinje/fisiologia , Ratos , Proteínas SNARE/metabolismo , Sinaptotagmina I/metabolismo
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