RESUMO
The double-stranded RNA-activated protein kinase (PKR) was originally identified as a sensor of virus infection, but its function in the brain remains unknown. Here, we report that the lack of PKR enhances learning and memory in several behavioral tasks while increasing network excitability. In addition, loss of PKR increases the late phase of long-lasting synaptic potentiation (L-LTP) in hippocampal slices. These effects are caused by an interferon-γ (IFN-γ)-mediated selective reduction in GABAergic synaptic action. Together, our results reveal that PKR finely tunes the network activity that must be maintained while storing a given episode during learning. Because PKR activity is altered in several neurological disorders, this kinase presents a promising new target for the treatment of cognitive dysfunction. As a first step in this direction, we show that a selective PKR inhibitor replicates the Pkr(-/-) phenotype in WT mice, enhancing long-term memory storage and L-LTP.
Assuntos
Hipocampo/fisiologia , Interferon gama/metabolismo , Potenciação de Longa Duração , eIF-2 Quinase/antagonistas & inibidores , eIF-2 Quinase/metabolismo , Animais , Eletrofisiologia , Técnicas In Vitro , Interferon gama/genética , Camundongos , Camundongos da Linhagem 129 , Camundongos Endogâmicos C57BL , Sinapses , eIF-2 Quinase/genéticaRESUMO
Atrial fibrillation (AF) is the most commonly diagnosed arrhythmia, and ECG remains the gold standard for diagnosing AF. Wrist-worn technologies are appealing for their ability to passively process near-continuous pulse signals. The clinical application of wearable devices is controversial. Our systematic review and meta-analysis qualitatively and quantitatively analyze available literature on wrist-worn wearable devices (Apple Watch, Samsung, and KardiaBand) and their sensitivity and specificity in detecting AF compared to conventional methods. Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were followed, yielding nine studies (n = 1,581). Observational studies assessing the sensitivity and specificity of wrist-worn wearables in detecting AF in patients with and without a history of AF were included and analyzed using a fixed-effect model with an inverse-variance method. In patients with a history of AF, the overall sensitivity between device groups did not significantly differ (96.83%; P = 0.207). Specificity significantly differed between Apple, Samsung, and KardiaBand (99.61%, 81.13%, and 97.98%, respectively; P<0.001). The effect size for this analysis was highest in the Samsung device group. Two studies (n = 796) differentiated cohorts to assess device sensitivity in patients with known AF and device specificity in patients with normal sinus rhythm (NSR) (sensitivity: 96.02%; confidence intervals (CI) 93.85%-97.59% and specificity: 98.82%; CI:97.46%-99.57%). Wrist-worn wearable devices demonstrate promising results in detecting AF in patients with paroxysmal AF. However, more rigorous prospective data is needed to understand the limitations of these devices in regard to varying specificities which may lead to unintended downstream medical testing and costs.
RESUMO
S 24795 evoked methyllycaconitine-sensitive inward currents in voltage-clamped hippocampal interneurons with maximum amplitude about 14% that of ACh-evoked responses. Experiments with rat alpha7 receptors expressed in Xenopus oocytes confirmed that S 24795 is a partial agonist of alpha7 nAChR with an EC(50) of 34+/-11 microM and I(max) of approximately 10% relative to ACh. When 60 microM ACh was co-applied to alpha7-expressing oocytes along with increasing concentrations of S 24795, there was a progressive decrease in response compared to the responses to 60 microM ACh alone (IC(50) 45+/-9 microM). The positive allosteric modulator 5-hydroxyindole potentiated ACh- and S 24795-evoked responses of alpha7 receptors in both oocytes and hippocampal interneurons. In hippocampal slice experiments, depending on the ACh concentrations in the application pipette and the ratio of ACh to S 24795, co-application of S 24795 with ACh variously increased, decreased, or had no effect on responses, compared to ACh alone. In order to estimate the effective dilution factor for the pressure application experiments, we tested alpha7 receptors in oocytes with ACh alone and in co-application with S 24795 at the same ratios as in the slice experiments, but at varying dilution factors. The pattern of interaction seen in the slice experiments was most closely matched under the conditions of a 3:100 dilution, suggesting that the pipette solution was diluted approximately 30-fold at the site of action. This dilution factor was consistent with the potency of ACh and S 24795 in the oocyte expression system (EC(50)s approximately 30 microM).
Assuntos
Agonistas Colinérgicos/farmacologia , Hipocampo/citologia , Interneurônios/efeitos dos fármacos , Receptores Nicotínicos/fisiologia , Acetilcolina/farmacologia , Aconitina/análogos & derivados , Aconitina/farmacologia , Animais , Animais Recém-Nascidos , Antagonistas Colinérgicos/farmacologia , Relação Dose-Resposta a Droga , Interações Medicamentosas , Feminino , Técnicas In Vitro , Indóis/farmacologia , Concentração Inibidora 50 , Masculino , Potenciais da Membrana/efeitos dos fármacos , Potenciais da Membrana/fisiologia , Potenciais da Membrana/efeitos da radiação , Oócitos , Técnicas de Patch-Clamp , Compostos de Piridínio/farmacologia , Ratos , Ratos Sprague-Dawley , Xenopus , Receptor Nicotínico de Acetilcolina alfa7RESUMO
Nicotinic acetylcholine receptors (nAChRs) are widely expressed throughout the central nervous system and participate in a variety of physiological functions. Recent advances have revealed roles of nAChRs in the regulation of synaptic transmission and synaptic plasticity, particularly in the hippocampus and midbrain dopamine centers. In general, activation of nAChRs causes membrane depolarization and directly and indirectly increases the intracellular calcium concentration. Thus, when nAChRs are expressed on presynaptic membranes their activation generally increases the probability of neurotransmitter release. When expressed on postsynaptic membranes, nAChR-initiated calcium signals and depolarization activate intracellular signaling mechanisms and gene transcription. Together, the presynaptic and postsynaptic effects of nAChRs generate and facilitate the induction of long-term changes in synaptic transmission. The direction of hippocampal nAChR-mediated synaptic plasticity - either potentiation or depression - depends on the timing of nAChR activation relative to coincident presynaptic and postsynaptic electrical activity, and also depends on the location of cholinergic stimulation within the local network. Therapeutic activation of nAChRs may prove efficacious in the treatment of neuropathologies where synaptic transmission is compromised, as in Alzheimer's or Parkinson's disease.
Assuntos
Hipocampo/fisiologia , Plasticidade Neuronal , Receptores Nicotínicos/fisiologia , Transmissão Sináptica , Animais , Humanos , Potenciação de Longa Duração , Receptores Nicotínicos/química , Receptores Nicotínicos/efeitos dos fármacos , Transdução de Sinais , Área Tegmentar Ventral/fisiologiaRESUMO
Adolescents are especially prone to drug addiction, but the underlying biological basis of their increased vulnerability remains unknown. We reveal that translational control by phosphorylation of the translation initiation factor eIF2α (p-eIF2α) accounts for adolescent hypersensitivity to cocaine. In adolescent (but not adult) mice, a low dose of cocaine reduced p-eIF2α in the ventral tegmental area (VTA), potentiated synaptic inputs to VTA dopaminergic neurons, and induced drug-reinforced behavior. Like adolescents, adult mice with reduced p-eIF2α-mediated translational control were more susceptible to cocaine-induced synaptic potentiation and behavior. Conversely, like adults, adolescent mice with increased p-eIF2α became more resistant to cocaine's effects. Accordingly, metabotropic glutamate receptor-mediated long-term depression (mGluR-LTD)-whose disruption is postulated to increase vulnerability to drug addiction-was impaired in both adolescent mice and adult mice with reduced p-eIF2α mediated translation. Thus, during addiction, cocaine hijacks translational control by p-eIF2α, initiating synaptic potentiation and addiction-related behaviors. These insights may hold promise for new treatments for addiction.
Assuntos
Comportamento/efeitos dos fármacos , Cocaína/metabolismo , Fator de Iniciação 2 em Eucariotos/metabolismo , Biossíntese de Proteínas , Processamento de Proteína Pós-Traducional , Sinapses/efeitos dos fármacos , Área Tegmentar Ventral/fisiologia , Animais , Camundongos , FosforilaçãoRESUMO
Recreational drug use leads to compulsive substance abuse in some individuals. Studies on animal models of drug addiction indicate that persistent long-term potentiation (LTP) of excitatory synaptic transmission onto ventral tegmental area (VTA) dopamine (DA) neurons is a critical component of sustained drug seeking. However, little is known about the mechanism regulating such long-lasting changes in synaptic strength. Previously, we identified that translational control by eIF2α phosphorylation (p-eIF2α) regulates cocaine-induced LTP in the VTA (Huang et al., 2016). Here we report that in mice with reduced p-eIF2α-mediated translation, cocaine induces persistent LTP in VTA DA neurons. Moreover, selectively inhibiting eIF2α-mediated translational control with a small molecule ISRIB, or knocking down oligophrenin-1-an mRNA whose translation is controlled by p-eIF2α-in the VTA also prolongs cocaine-induced LTP. This persistent LTP is mediated by the insertion of GluR2-lacking AMPARs. Collectively, our findings suggest that eIF2α-mediated translational control regulates the progression from transient to persistent cocaine-induced LTP.
Assuntos
Potenciais de Ação/efeitos dos fármacos , Cocaína/metabolismo , Inibidores da Captação de Dopamina/metabolismo , Neurônios Dopaminérgicos/fisiologia , Fator de Iniciação 2 em Eucariotos/metabolismo , Biossíntese de Proteínas , Área Tegmentar Ventral/fisiologia , Animais , Neurônios Dopaminérgicos/efeitos dos fármacos , Camundongos , Área Tegmentar Ventral/efeitos dos fármacosRESUMO
Adolescents are particularly vulnerable to nicotine, the principal addictive component driving tobacco smoking. In a companion study, we found that reduced activity of the translation initiation factor eIF2α underlies the hypersensitivity of adolescent mice to the effects of cocaine. Here we report that nicotine potentiates excitatory synaptic transmission in ventral tegmental area dopaminergic neurons more readily in adolescent mice compared to adults. Adult mice with genetic or pharmacological reduction in p-eIF2α-mediated translation are more susceptible to nicotine's synaptic effects, like adolescents. When we investigated the influence of allelic variability of the Eif2s1 gene (encoding eIF2α) on reward-related neuronal responses in human smokers, we found that a single nucleotide polymorphism in the Eif2s1 gene modulates mesolimbic neuronal reward responses in human smokers. These findings suggest that p-eIF2α regulates synaptic actions of nicotine in both mice and humans, and that reduced p-eIF2α may enhance susceptibility to nicotine (and other drugs of abuse) during adolescence.
Assuntos
Neurônios Dopaminérgicos/fisiologia , Fator de Iniciação 2 em Eucariotos/metabolismo , Nicotina/metabolismo , Biossíntese de Proteínas , Processamento de Proteína Pós-Traducional , Sinapses/efeitos dos fármacos , Área Tegmentar Ventral/fisiologia , Animais , Humanos , Camundongos , Fosforilação , Fumar , NicotianaRESUMO
At hippocampal synapses, activation of group I metabotropic glutamate receptors (mGluRs) induces long-term depression (LTD), which requires new protein synthesis. However, the underlying mechanism remains elusive. Here we describe the translational program that underlies mGluR-LTD and identify the translation factor eIF2α as its master effector. Genetically reducing eIF2α phosphorylation, or specifically blocking the translation controlled by eIF2α phosphorylation, prevented mGluR-LTD and the internalization of surface AMPA receptors (AMPARs). Conversely, direct phosphorylation of eIF2α, bypassing mGluR activation, triggered a sustained LTD and removal of surface AMPARs. Combining polysome profiling and RNA sequencing, we identified the mRNAs translationally upregulated during mGluR-LTD. Translation of one of these mRNAs, oligophrenin-1, mediates the LTD induced by eIF2α phosphorylation. Mice deficient in phospho-eIF2α-mediated translation are impaired in object-place learning, a behavioral task that induces hippocampal mGluR-LTD in vivo. Our findings identify a new model of mGluR-LTD, which promises to be of value in the treatment of mGluR-LTD-linked cognitive disorders.
Assuntos
Fator de Iniciação 2 em Eucariotos/genética , Fator de Iniciação 2 em Eucariotos/metabolismo , Aprendizagem/fisiologia , Depressão Sináptica de Longo Prazo/genética , Biossíntese de Proteínas , Receptores de AMPA/fisiologia , Receptores de Glutamato Metabotrópico/metabolismo , Animais , Regulação da Expressão Gênica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Fosforilação/genética , Receptores de AMPA/deficiência , Percepção Espacial/fisiologiaRESUMO
Dopamine (DA) neurons in the ventral tegmental area (VTA) have been implicated in brain mechanisms related to motivation, reward, and drug addiction. Successful identification of these neurons in vitro has historically depended upon the expression of a hyperpolarization-activated current (I(h)) and immunohistochemical demonstration of the presence of tyrosine hydroxylase (TH), the rate-limiting enzyme for DA synthesis. Recent findings suggest that electrophysiological criteria may be insufficient for distinguishing DA neurons from non-DA neurons in the VTA. In this study, we sought to determine factors that could potentially account for the apparent discrepancies in the literature regarding DA neuron identification in the rodent brain slice preparation. We found that confirmed DA neurons from the lateral VTA generally displayed a larger amplitude I(h) relative to DA neurons located in the medial VTA. Measurement of a large amplitude I(h) (>100 pA) consistently indicated a dopaminergic phenotype, but non-dopamine neurons also can have I(h) current. The data also showed that immunohistochemical TH labeling of DA neurons can render false negative results after relatively long duration (>15 min) whole-cell patch clamp recordings. We conclude that whole-cell patch clamp recording in combination with immunohistochemical detection of TH expression can guarantee positive but not negative DA identification in the VTA.
Assuntos
Dopamina/metabolismo , Neurônios/fisiologia , Área Tegmentar Ventral/fisiologia , Potenciais de Ação/fisiologia , Análise de Variância , Animais , Eletrofisiologia , Imuno-Histoquímica , Camundongos , Camundongos Transgênicos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Tirosina 3-Mono-Oxigenase/metabolismoRESUMO
The dopamine (DA) system of the ventral midbrain plays a critical role as mammals learn adaptive behaviors driven by environmental salience and reward. Addictive drugs, including nicotine, exert powerful influences over the mesolimbic DA system by activating and desensitizing nicotinic acetylcholine receptors (nAChRs) in a subtype-dependent manner. Nicotine induces synaptic plasticity at excitatory synapses onto DA neurons, thereby sending elevated DA signals that participate during the reinforcement of addictive behaviors. While humans and animals of any developmental age are potentially vulnerable to these drug-induced effects, evidence from clinical and epidemiological studies indicates that adolescents have an increased risk of addiction. Although this risk arises from a complex set of variables including societal and psychosocial influences, a contributing factor involves age dependent sensitivity to addictive drugs. One aspect of that sensitivity is drug-induced synaptic plasticity at excitatory synapses onto the dopamine neurons in the ventral midbrain. A single, acute exposure to addictive drugs, including nicotine, produces long-term potentiation (LTP) that can be quantified by measuring the shift in the subtypes of ionotropic glutamate receptors mediating evoked synaptic transmission. This change in glutamatergic transmission is expressed as an increased ratio of AMPA receptors to NMDA receptors at glutamatergic synapses. Age-related differences in the excitability and the nicotine sensitivity within the midbrain dopamine system may contribute to the greater risk of nicotine addiction in adolescent animals and humans.
Assuntos
Dopamina/fisiologia , Plasticidade Neuronal/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Nicotina/farmacologia , Sinapses/efeitos dos fármacos , Adolescente , Comportamento do Adolescente , Fatores Etários , Animais , Humanos , Mesencéfalo/efeitos dos fármacos , Mesencéfalo/crescimento & desenvolvimento , Mesencéfalo/fisiopatologia , Neurônios/fisiologia , Nicotina/efeitos adversos , Receptores Nicotínicos/fisiologia , Sinapses/fisiologia , Tabagismo/fisiopatologia , Tabagismo/prevenção & controleRESUMO
The alpha7-type nicotinic acetylcholine receptor (nAChR) has been recognized as a potential therapeutic target for the treatment of a variety of pathologic conditions, including schizophrenia, Alzheimer's disease, and peripheral inflammation. A unique feature of alpha7 nAChRs that tends to complicate functional assays intended to identify selective drugs for these receptors is the strong concentration-dependent desensitization of their agonist-evoked responses. At low agonist concentrations, voltage-clamp responses are small but tend to closely follow the solution exchange profile, whereas higher agonist concentrations produce responses that peak and then decay very rapidly, usually before the full drug concentration has been achieved. In this article, we report that an alpha7 T245S mutant, which has a point mutation at the sixth position in the alpha7 second transmembrane domain (T6'S), demonstrates a significant gain of function, sustaining current when exposed to relatively high agonist concentrations when expressed in Xenopus laevis oocytes and larger peak currents when expressed in mammalian GH4C1 cells. At the single-channel level, the T6'S mutant has a unitary conductance of 61.7 +/- 5.8 pS, similar to that reported for wild-type alpha7, but a vastly longer average open duration. In addition, channel burst activity indicates a greater than 40% probability of channel re-opening in the sustained presence of 30 muM acetylcholine, consistent with a greater overall open probability relative to wild-type alpha7. Unlike the alpha7 L248T gain-of-function mutant, the T6'S mutant exhibits a pharmacological profile that is remarkably similar to the wild-type alpha7 receptor, implicating it as a potentially useful tool for identifying therapeutic agents.
Assuntos
Mutação de Sentido Incorreto , Receptores Nicotínicos/genética , Animais , Cálcio/metabolismo , Eletrofisiologia , Cinética , Mutagênese Sítio-Dirigida , Oócitos , Técnicas de Patch-Clamp , Ratos , Receptores Nicotínicos/metabolismo , Receptores Nicotínicos/fisiologia , Especificidade por Substrato , Transfecção , Xenopus laevis , Receptor Nicotínico de Acetilcolina alfa7Assuntos
Dopamina/metabolismo , Mesencéfalo/efeitos dos fármacos , Mesencéfalo/fisiopatologia , Motivação , Plasticidade Neuronal/efeitos dos fármacos , Plasticidade Neuronal/fisiologia , Nicotina/farmacologia , Fumar/fisiopatologia , Sinapses/efeitos dos fármacos , Sinapses/fisiologia , Tabagismo/fisiopatologia , Animais , Humanos , Potenciação de Longa Duração/efeitos dos fármacos , Potenciação de Longa Duração/fisiologia , Nicotina/toxicidade , Receptores Nicotínicos/efeitos dos fármacos , Receptores Nicotínicos/fisiologia , Fumar/psicologia , Síndrome de Abstinência a Substâncias/fisiopatologia , Síndrome de Abstinência a Substâncias/psicologia , Tabagismo/psicologia , Adulto JovemRESUMO
Although the muscle-type and homomeric alpha7-type nicotinic acetylcholine receptors (nAChRs) share many structural features and bind alpha-bungarotoxin with high affinity, several important functional and pharmacological properties distinguish these two major nAChR subtypes. We have shown previously that amino acid sequence in the second transmembrane (TM) domain of the beta subunit is critical for pharmacological distinction between muscle type and heteromeric neuronal (e.g., ganglionic) nAChRs. We tested the hypothesis that homologous substitution of amino acid sequence from the muscle beta1 subunit into the alpha7 subunit would confer specific properties of muscle-type receptors to mutant alpha7 nAChRs. In this study, we show that a single amino acid substitution at the alpha7 TM2 6' position makes both biophysical and pharmacological properties of the mutant receptors resemble those of wild-type muscle nAChR. This mutation produces significant changes in acetylcholine potency and response kinetics, eliminating the characteristic fast desensitization of alpha7 and dramatically reducing divalent ion permeability relative to wild-type alpha7. The TM2 T6'F mutation also produces a profound increase in activation by succinylcholine compared with either wild-type alpha7 or neuronal beta-subunit-containing receptors and the loss of potentiation by 5-hydroxyindole. Thus, the alpha7 TM2 T6'F mutant displays several features that are similar to the muscle nAChR, some of which are not typically thought to be regulated by the pore-lining domain of the receptor.