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In this issue, Papouin et al. show that glycine is the endogenous coagonist for extrasynaptic NMDA receptors (NMDARs), unlike at synapses where the coagonist is d-serine. By enzymatically degrading endogenous glycine, they begin to address the enigmatic physiological and pathological roles for extrasynaptic NMDARs.
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NMDA-type glutamate receptors (NMDARs) are widely recognized as master regulators of synaptic plasticity, most notably for driving long-term changes in synapse size and strength that support learning. NMDARs are unique among neurotransmitter receptors in that they require binding of both neurotransmitter (glutamate) and co-agonist (e.g., d-serine) to open the receptor channel, which leads to the influx of calcium ions that drive synaptic plasticity. Over the past decade, evidence has accumulated that NMDARs also support synaptic plasticity via ion flux-independent (non-ionotropic) signaling upon the binding of glutamate in the absence of co-agonist, although conflicting results have led to significant controversy. Here, we hypothesized that a major source of contradictory results might be attributed to variable occupancy of the co-agonist binding site under different experimental conditions. To test this hypothesis, we manipulated co-agonist availability in acute hippocampal slices from mice of both sexes. We found that enzymatic scavenging of endogenous co-agonists enhanced the magnitude of long-term depression (LTD) induced by non-ionotropic NMDAR signaling in the presence of the NMDAR pore blocker MK801. Conversely, a saturating concentration of d-serine completely inhibited LTD and spine shrinkage induced by glutamate binding in the presence of MK801 or Mg2+ Using a Förster resonance energy transfer (FRET)-based assay in cultured neurons, we further found that d-serine completely blocked NMDA-induced conformational movements of the GluN1 cytoplasmic domains in the presence of MK801. Our results support a model in which d-serine availability serves to modulate NMDAR signaling and synaptic plasticity even when the NMDAR is blocked by magnesium.
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Hipocampo , Receptores de N-Metil-D-Aspartato , Serina , Transducción de Señal , Animales , Receptores de N-Metil-D-Aspartato/metabolismo , Ratones , Masculino , Femenino , Serina/metabolismo , Serina/farmacología , Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Ratones Endogámicos C57BL , Depresión Sináptica a Largo Plazo/efectos de los fármacos , Depresión Sináptica a Largo Plazo/fisiología , Ácido Glutámico/metabolismo , Plasticidad Neuronal/efectos de los fármacos , Plasticidad Neuronal/fisiología , Neuronas/efectos de los fármacos , Neuronas/metabolismoRESUMEN
Insect nervous systems offer unique advantages for studying interactions between sensory systems and behavior, given their complexity with high tractability. By examining the neural coding of salient environmental stimuli and resulting behavioral output in the context of environmental stressors, we gain an understanding of the effects of these stressors on brain and behavior and provide insight into normal function. The implication of neonicotinoid (neonic) pesticides in contributing to declines of nontarget species, such as bees, has motivated the development of new compounds that can potentially mitigate putative resistance in target species and declines of nontarget species. We used a neuroethologic approach, including behavioral assays and multineuronal recording techniques, to investigate effects of imidacloprid (IMD) and the novel insecticide sulfoxaflor (SFX) on visual motion-detection circuits and related escape behavior in the tractable locust system. Despite similar LD50 values, IMD and SFX evoked different behavioral and physiological effects. IMD significantly attenuated collision avoidance behaviors and impaired responses of neural populations, including decreases in spontaneous firing and neural habituation. In contrast, SFX displayed no effect at a comparable sublethal dose. These results show that neonics affect population responses and habituation of a visual motion detection system. We propose that differences in the sublethal effects of SFX reflect a different mode of action than that of IMD. More broadly, we suggest that neuroethologic assays for comparative neurotoxicology are valuable tools for fully addressing current issues regarding the proximal effects of environmental toxicity in nontarget species.
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Exposición a Riesgos Ambientales , Reacción de Fuga/efectos de los fármacos , Insecticidas/toxicidad , Neuronas Motoras/efectos de los fármacos , Neonicotinoides/toxicidad , Nitrocompuestos/toxicidad , Piridinas/toxicidad , Compuestos de Azufre/toxicidad , Animales , Habituación Psicofisiológica/efectos de los fármacos , Dosificación Letal Mediana , Locusta migratoria/efectos de los fármacos , Movimiento (Física)RESUMEN
The quality of drug products in the United States has been a matter of growing concern. Buyers and payers of pharmaceuticals have limited insight into measures of drug-product quality. Therefore, a quality-score system driven by data collection is proposed to differentiate between the qualities of drug products produced by different manufacturers. The quality scores derived using this proposed system would be based upon public regulatory data and independently-derived chemical data. A workflow for integrating the system into procurement decisions within health care organizations is also suggested. The implementation of such a quality-score system would benefit health care organizations by including the consideration of the quality of products while also considering price as a part of the drug procurement process. Such a system would also benefit the U.S. health care industry by bringing accountability and transparency into the drug supply chain and incentivizing manufacturers to place an increased emphasis on the quality and safety of their drug products.
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Industria Farmacéutica , Sector de Atención de Salud , Humanos , Estados UnidosRESUMEN
Angelman syndrome (AS) is a rare genetic neurodevelopmental disorder characterized by intellectual disabilities, motor and balance deficits, impaired communication, and a happy, excitable demeanor with frequent laughter. We sought to elucidate a preclinical outcome measure in male and female rats that addressed communication abnormalities of AS and other neurodevelopmental disorders in which communication is atypical and/or lack of speech is a core feature. We discovered, and herein report for the first time, excessive laughter-like 50 kHz ultrasonic emissions in the Ube3amat-/pat+ rat model of AS, which suggests an excitable, playful demeanor and elevated positive affect, similar to the demeanor of individuals with AS. Also in line with the AS phenotype, Ube3amat-/pat+ rats demonstrated aberrant social interactions with a novel partner, distinctive gait abnormalities, impaired cognition, an underlying LTP deficit, and profound reductions in brain volume. These unique, robust phenotypes provide advantages compared with currently available mouse models and will be highly valuable as outcome measures in the evaluation of therapies for AS.SIGNIFICANCE STATEMENT Angelman syndrome (AS) is a severe neurogenetic disorder for which there is no cure, despite decades of research using mouse models. This study used a recently developed rat model of AS to delineate disease-relevant outcome measures to facilitate therapeutic development. We found the rat to be a strong model of AS, offering several advantages over mouse models by exhibiting numerous AS-relevant phenotypes, including overabundant laughter-like vocalizations, reduced hippocampal LTP, and volumetric anomalies across the brain. These findings are unconfounded by detrimental motor abilities and background strain, issues plaguing mouse models. This rat model represents an important advancement in the field of AS, and the outcome metrics reported herein will be central to the therapeutic pipeline.
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Síndrome de Angelman/genética , Modelos Animales de Enfermedad , Risa/fisiología , Microcefalia/genética , Ubiquitina-Proteína Ligasas/genética , Vocalización Animal/fisiología , Síndrome de Angelman/metabolismo , Síndrome de Angelman/psicología , Animales , Encéfalo/metabolismo , Femenino , Eliminación de Gen , Risa/psicología , Masculino , Microcefalia/metabolismo , Microcefalia/psicología , Técnicas de Cultivo de Órganos , Biosíntesis de Proteínas/fisiología , Ratas , Ratas Sprague-Dawley , Ratas Transgénicas , Reflejo de Sobresalto/fisiología , Conducta Social , Ubiquitina-Proteína Ligasas/deficienciaRESUMEN
Schizophrenia is a psychiatric disorder that affects over 20 million people globally. Notably, schizophrenia is associated with decreased density of dendritic spines and decreased levels of d-serine, a co-agonist required for opening of the N-methyl-d-aspartate receptor (NMDAR). We hypothesized that lowered d-serine levels associated with schizophrenia would enhance ion flux-independent signaling by the NMDAR, driving destabilization and loss of dendritic spines. We tested our hypothesis using the serine racemase knockout (SRKO) mouse model, which lacks the enzyme for d-serine production. We show that activity-dependent spine growth is impaired in SRKO mice, but can be acutely rescued by exogenous d-serine. Moreover, we find a significant bias of synaptic plasticity toward spine shrinkage in the SRKO mice as compared to wild-type littermates. Notably, we demonstrate that enhanced ion flux-independent signaling through the NMDAR contributes to this bias toward spine destabilization, which is exacerbated by an increase in synaptic NMDARs in hippocampal synapses of SRKO mice. Our results support a model in which lowered d-serine levels associated with schizophrenia enhance ion flux-independent NMDAR signaling and bias toward spine shrinkage and destabilization.
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Receptores de N-Metil-D-Aspartato , Esquizofrenia , Animales , Espinas Dendríticas , Modelos Animales de Enfermedad , Humanos , Ratones , Ratones Noqueados , Plasticidad Neuronal , SerinaRESUMEN
We present a follow-on experiment to the recent study from The University of Rochester [Opt. Lett.42, 2479 (2017)OPLEDP0146-959210.1364/OL.42.002479], which reported a new architecture for an inverse weak-value tiltmeter. We recreate the Rochester tiltmeter and specifically investigate mirror oscillations in the low-kilohertz frequency regime, which is relevant to certain potential applications, such as Coriolis vibratory gyroscopes. We find that the inverse weak-value amplification effect persists in this regime, although our measured noise floors are higher than those obtained in the Rochester experiment-approximately 2prad/Hz for mirror oscillation frequencies between 1 and 25 kHz.
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d-serine is the primary NMDAR coagonist at mature forebrain synapses and is synthesized by the enzyme serine racemase (SR). However, our understanding of the mechanisms regulating the availability of synaptic d-serine remains limited. Though early studies suggested d-serine is synthesized and released from astrocytes, more recent studies have demonstrated a predominantly neuronal localization of SR. More specifically, recent work intriguingly suggests that SR may be found at the postsynaptic density, yet the functional implications of postsynaptic SR on synaptic transmission are not yet known. Here, we show an age-dependent dendritic and postsynaptic localization of SR and d-serine by immunohistochemistry and electron microscopy in mouse CA1 pyramidal neurons. In addition, using a single-neuron genetic approach in SR conditional KO mice from both sexes, we demonstrate a cell-autonomous role for SR in regulating synaptic NMDAR function at Schaffer collateral (CA3)-CA1 synapses. Importantly, single-neuron genetic deletion of SR resulted in the elimination of LTP at 1 month of age, which could be rescued by exogenous d-serine. Interestingly, there was a restoration of LTP by 2 months of age that was associated with an upregulation of synaptic GluN2B. Our findings support a cell-autonomous role for postsynaptic neuronal SR in regulating synaptic NMDAR function and suggests a possible autocrine mode of d-serine action.SIGNIFICANCE STATEMENT NMDARs are key regulators of neurodevelopment and synaptic plasticity and are unique in their requirement for binding of a coagonist, which is d-serine at most forebrain synapses. However, our understanding of the mechanisms regulating synaptic d-serine availability remains limited. d-serine is synthesized in the brain by the neuronal enzyme serine racemase (SR). Here, we show dendritic and postsynaptic localization of SR and d-serine in CA1 pyramidal neurons. In addition, using single-neuron genetic deletion of SR, we establish a role of postsynaptic SR in regulating NMDAR function. These results support an autocrine mode of d-serine action at synapses.
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Dendritas/metabolismo , Células Piramidales/metabolismo , Racemasas y Epimerasas/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Sinapsis/metabolismo , Factores de Edad , Animales , Región CA1 Hipocampal/metabolismo , Femenino , Masculino , Ratones , Ratones Noqueados , Plasticidad Neuronal/fisiología , Racemasas y Epimerasas/genética , Transmisión Sináptica/fisiologíaRESUMEN
There is substantial evidence that both N-methyl-D-aspartate receptor (NMDAR) hypofunction and dysfunction of GABAergic neurotransmission contribute to schizophrenia, though the relationship between these pathophysiological processes remains largely unknown. Although models using cell-type-specific genetic deletion of NMDARs have been informative, they display overly pronounced phenotypes extending beyond those of schizophrenia. Here, we used the serine racemase knockout (SRKO) mice, a model of reduced NMDAR activity rather than complete receptor elimination, to examine the link between NMDAR hypofunction and decreased GABAergic inhibition. The SRKO mice, in which there is a >90% reduction in the NMDAR coagonist d-serine, exhibit many of the neurochemical and behavioral abnormalities observed in schizophrenia. We found a significant reduction in inhibitory synapses onto CA1 pyramidal neurons in the SRKO mice. This reduction increases the excitation/inhibition balance resulting in enhanced synaptically driven neuronal excitability without changes in intrinsic excitability. Consistently, significant reductions in inhibitory synapse density in CA1 were observed by immunohistochemistry. We further show, using a single-neuron genetic deletion approach, that the loss of GABAergic synapses onto pyramidal neurons observed in the SRKO mice is driven in a cell-autonomous manner following the deletion of SR in individual CA1 pyramidal cells. These results support a model whereby NMDAR hypofunction in pyramidal cells disrupts GABAergic synapses leading to disrupted feedback inhibition and impaired neuronal synchrony.NEW & NOTEWORTHY Recently, disruption of excitation/inhibition (E/I) balance has become an area of considerable interest for psychiatric research. Here, we report a reduction in inhibition in the serine racemase knockout mouse model of schizophrenia that increases E/I balance and enhances synaptically driven neuronal excitability. This reduced inhibition was driven cell-autonomously in pyramidal cells lacking serine racemase, suggesting a novel mechanism for how chronic NMDA receptor hypofunction can disrupt information processing in schizophrenia.
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Potenciales Postsinápticos Excitadores/fisiología , Neuronas GABAérgicas/metabolismo , Potenciales Postsinápticos Inhibidores/fisiología , Racemasas y Epimerasas/deficiencia , Receptores de N-Metil-D-Aspartato/deficiencia , Sinapsis/metabolismo , Animales , Femenino , Hipocampo/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Técnicas de Cultivo de Órganos , Racemasas y Epimerasas/genética , Receptores de N-Metil-D-Aspartato/genética , Esquizofrenia/genética , Esquizofrenia/metabolismo , Sinapsis/genéticaRESUMEN
Detection of looming obstacles is a vital task for both natural and artificial systems. Locusts possess a visual nervous system with an extensively studied obstacle detection pathway, culminating in the lobula giant movement detector (LGMD) neuron. While numerous models of this system exist, none to date have incorporated recent data on the anatomy and function of feedforward and global inhibitory systems in the input network of the LGMD. Moreover, the possibility that global and lateral inhibition shape the feedforward inhibitory signals to the LGMD has not been investigated. To address these points, a novel model of feedforward inhibitory neurons in the locust optic lobe was developed based on the recent literature. This model also incorporated global and lateral inhibition into the afferent network of these neurons, based on their observed behaviour in existing data and the posited role of these mechanisms in the inputs to the LGMD. Tests with the model showed that it accurately replicates the behaviour of feedforward inhibitory neurons in locusts; the model accurately coded for stimulus angular size in an overall linear fashion, with decreasing response saturation and increasing linearity as stimulus size increased or approach velocity decreased. The model also exhibited only phasic responses to the appearance of a grating, along with sustained movement by it at constant speed. By observing the effects of altering inhibition schemes on these responses, it was determined that global inhibition serves primarily to normalize growing excitation as collision approaches, and keeps coding for subtense angle linear. Lateral inhibition was determined to suppress tonic responses to wide-field stimuli translating at constant speed. Based on these features being shared with characterizations of the LGMD input network, it was hypothesized that the feedforward inhibitory neurons and the LGMD share the same excitatory afferents; this necessitates further investigation.
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Saltamontes , Percepción de Movimiento , AnimalesRESUMEN
Multiple clinical trials employing recombinant adeno-associated viral (rAAV) vectors have been initiated for neuromuscular disorders, including Duchenne and limb-girdle muscular dystrophies, spinal muscular atrophy, and recently X-linked myotubular myopathy (XLMTM). Our previous work on a canine model of XLMTM showed that a single rAAV8-cMTM1 systemic infusion corrected structural abnormalities within the muscle and restored contractile function, with affected dogs surviving more than 4 years post injection. This remarkable therapeutic efficacy presents a unique opportunity to identify the downstream molecular drivers of XLMTM pathology and to what extent the whole muscle transcriptome is restored to normal after gene transfer. Herein, RNA-sequencing was used to examine the transcriptomes of the Biceps femoris and Vastus lateralis in a previously described canine cohort that showed dose-dependent clinical improvements after rAAV8-cMTM1 gene transfer. Our analysis confirmed several dysregulated genes previously observed in XLMTM mice but also identified transcripts linked to XLMTM pathology. We demonstrated XLMTM transcriptome remodeling and dose-dependent normalization of gene expression after gene transfer and created metrics to pinpoint potential biomarkers of disease progression and correction.
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Dependovirus/genética , Técnicas de Transferencia de Gen , Terapia Genética , Vectores Genéticos/genética , Músculo Esquelético/metabolismo , Miopatías Estructurales Congénitas/genética , Transcriptoma , Animales , Biomarcadores , Modelos Animales de Enfermedad , Perros , Dosificación de Gen , Perfilación de la Expresión Génica , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Transducción GenéticaRESUMEN
Vacuum ultraviolet (VUV) light at 118 nm has been shown to be a powerful tool to ionize molecules for various gas-phase chemical studies. A convenient table top source of 118 nm light can be produced by frequency tripling 355 nm light from a Nd:YAG laser in xenon gas. This process has a low efficiency, typically producing only nJ/pulse of VUV light. Simple models of the tripling process predict that the power of 118 nm light produced should increase quadratically with increasing xenon pressure. However, experimental 118 nm production has been observed to reach a maximum and then decrease to zero with increasing xenon pressure. Here, we describe the basic theory and experimental setup for producing 118 nm light and a new proposed model for the mechanism limiting the production based on pressure broadened absorption.
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Aminoacyl-tRNA synthetases (AaRS) charge tRNAs with amino acids for protein translation. In plants, cytoplasmic, mitochondrial, and chloroplast AaRS exist that are all coded for by nuclear genes and must be imported from the cytosol. In addition, only a few of the mitochondrial tRNAs needed for translation are encoded in mitochondrial DNA. Despite considerable progress made over the last few years, still little is known how the bulk of cytosolic AaRS and respective tRNAs are transported into mitochondria. Here, we report the identification of a protein complex that ties AaRS and tRNA import into the mitochondria of Arabidopsis thaliana. Using leucyl-tRNA synthetase 2 (LeuRS2) as a model for a mitochondrial signal peptide (MSP)-less precursor, a ≈30 kDa protein was identified that interacts with LeuRS2 during import. The protein identified is identical with a previously characterized mitochondrial protein designated HP30-2 (encoded by At3g49560) that contains a sterile alpha motif (SAM) similar to that found in RNA binding proteins. HP30-2 is part of a larger protein complex that contains with TIM22, TIM8, TIM9 and TIM10 four previously identified components of the translocase for MSP-less precursors. Lack of HP30-2 perturbed mitochondrial biogenesis and function and caused seedling lethality during greening, suggesting an essential role of HP30-2 in planta.
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Arabidopsis/fisiología , Leucina-ARNt Ligasa/metabolismo , Mitocondrias/genética , Mitocondrias/metabolismo , ARN de Transferencia/genética , Transporte Biológico , Proteínas Mitocondriales/genética , Proteínas Mitocondriales/metabolismo , Mutación , Biogénesis de Organelos , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Unión Proteica , ARN de Transferencia/metabolismoRESUMEN
We describe the natural history of 'Amish' nemaline myopathy (ANM), an infantile-onset, lethal disease linked to a pathogenic c.505G>T nonsense mutation of TNNT1, which encodes the slow fiber isoform of troponin T (TNNT1; a.k.a. TnT). The TNNT1 c.505G>T allele has a carrier frequency of 6.5% within Old Order Amish settlements of North America. We collected natural history data for 106 ANM patients born between 1923 and 2017. Over the last two decades, mean age of molecular diagnosis was 16 ± 27 days. TNNT1 c.505G>T homozygotes were normal weight at birth but failed to thrive by age 9 months. Presenting neonatal signs were axial hypotonia, hip and shoulder stiffness, and tremors, followed by progressive muscle weakness, atrophy and contractures. Affected children developed thoracic rigidity, pectus carinatum and restrictive lung disease during infancy, and all succumbed to respiratory failure by 6 years of age (median survival 18 months, range 0.2-66 months). Muscle histology from two affected children showed marked fiber size variation owing to both Type 1 myofiber smallness (hypotrophy) and Type 2 fiber hypertrophy, with evidence of nemaline rods, myofibrillar disarray and vacuolar pathology in both fiber types. The truncated slow TNNT1 (TnT) fragment (p.Glu180Ter) was undetectable in ANM muscle, reflecting its rapid proteolysis and clearance from sarcoplasm. Similar functional and histological phenotypes were observed in other human cohorts and two transgenic murine models (Tnnt1-/- and Tnnt1 c.505G>T). These findings have implications for emerging molecular therapies, including the suitably of TNNT1 gene replacement for newborns with ANM or other TNNT1-associated myopathies.
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Debilidad Muscular/genética , Músculo Esquelético/patología , Miopatías Nemalínicas/genética , Troponina T/genética , Amish/genética , Animales , Niño , Codón sin Sentido/genética , Femenino , Homocigoto , Humanos , Recién Nacido , Masculino , Ratones , Debilidad Muscular/diagnóstico , Debilidad Muscular/fisiopatología , Músculo Esquelético/metabolismo , Miopatías Nemalínicas/diagnóstico , Miopatías Nemalínicas/fisiopatología , Patología Molecular , Fenotipo , Isoformas de Proteínas/genéticaRESUMEN
SEC14 and Spectrin domain-1 (Sestd1) is a synapse protein that exhibits a striking shift from the presynaptic to postsynaptic space as neurons mature postnatally in the mouse hippocampus. Hippocampal pyramidal neurons from mice with global genetic deletion of Sestd1 have reduced dendrite arbors, spines, and excitatory synapses. Electrophysiologically this correlates with cell-autonomous reductions in both AMPA- and NMDA-excitatory postsynaptic currents in individual hippocampal neurons from which Sestd1 has been deleted in vivo. These neurodevelopmental and functional deficits are associated with increased activation of the Rho family GTPases Rac1 and RhoA. Co-immunoprecipitation and mass spectrometry reveal that the Breakpoint Cluster Region protein, a Rho GTPase activating protein (GAP), forms complexes with Sestd1 in brain tissue. This complements earlier findings that Sestd1 can also partner with other Rho family GAPs and guanine nucleotide exchange factors. Our findings demonstrate that Sestd1 is a developmentally dynamic synaptic regulator of Rho GTPases that contributes to dendrite and excitatory synapse formation within differentiating pyramidal neurons of the forebrain.
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Proteínas Portadoras/metabolismo , Espinas Dendríticas/metabolismo , Neuropéptidos/metabolismo , Prosencéfalo/metabolismo , Proteínas Proto-Oncogénicas c-bcr/metabolismo , Sinapsis/metabolismo , Proteína de Unión al GTP rac1/metabolismo , Animales , Proteínas Portadoras/análisis , Dendritas/química , Dendritas/metabolismo , Espinas Dendríticas/química , Femenino , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Neurogénesis/fisiología , Neuropéptidos/análisis , Técnicas de Cultivo de Órganos , Prosencéfalo/química , Prosencéfalo/crecimiento & desarrollo , Proteínas Proto-Oncogénicas c-bcr/análisis , Sinapsis/química , Proteína de Unión al GTP rac1/análisisRESUMEN
NMDA receptors (NMDARs) mediate both long-term potentiation and long-term depression (LTD) and understanding how a single receptor can initiate both phenomena remains a major question in neuroscience. A prominent hypothesis implicates the NMDAR subunit composition, specifically GluN2A and GluN2B, in dictating the rules of synaptic plasticity. However, studies testing this hypothesis have yielded inconsistent and often contradictory results, especially for LTD. These inconsistent results may be due to challenges in the interpretation of subunit-selective pharmacology and in dissecting out the contributions of differential channel properties versus the interacting proteins unique to GluN2A or GluN2B. In this study, we address the pharmacological and biochemical challenges by using a single-neuron genetic approach to delete NMDAR subunits in conditional knock-out mice. In addition, the recently discovered non-ionotropic nature of NMDAR-dependent LTD allowed the rigorous assessment of unique subunit contributions to NMDAR-dependent LTD while eliminating the variable of differential charge transfer. Here we find that neither the GluN2A nor the GluN2B subunit is strictly necessary for either non-ionotropic or ionotropic LTD.SIGNIFICANCE STATEMENT NMDA receptors are key regulators of bidirectional synaptic plasticity. Understanding the mechanisms regulating bidirectional plasticity will guide development of therapeutic strategies to treat the dysfunctional synaptic plasticity in multiple neuropsychiatric disorders. Because of the unique properties of the NMDA receptor GluN2 subunits, they have been postulated to differentially affect synaptic plasticity. However, there has been significant controversy regarding the roles of the GluN2 subunits in synaptic long term depression (LTD). Using single-neuron knock-out of the GluN2 subunits, we show that LTD requires neither GluN2A nor GluN2B.
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Depresión Sináptica a Largo Plazo/fisiología , Receptores de N-Metil-D-Aspartato/fisiología , Animales , Femenino , Masculino , Ratones , Ratones Noqueados , Resultados Negativos , Plasticidad Neuronal/fisiología , Neuronas/metabolismo , Receptores de Glutamato Metabotrópico/efectos de los fármacos , Receptores de Glutamato Metabotrópico/fisiología , Receptores de N-Metil-D-Aspartato/efectos de los fármacos , Receptores de N-Metil-D-Aspartato/metabolismoRESUMEN
A theory-driven tailored intervention developed in partnership with the community used evidence-based practices to (a) increase knowledge about colon cancer and screening and (b) assist patients in completing screenings. During the 16 months of delivery screening, patient navigators integrated into gastroenterology clinics met all goals, which included (a) enrolling an ethnically diverse group of participants ( N = 415) through inreach (clinic-referred patients who did not schedule appointments) and community outreach, (b) facilitating screening completions for 217 of the 358 (61%) patients identified as needing screening, and (c) obtaining satisfaction ratings from 89% of participants. A random sample ( N = 214) of nonnavigated patients matched on gender and age revealed no differences between navigated and nonnavigated patients on polyps detected. Navigated males (but not females) were significantly less likely than nonnavigated males to have either poor or only fair bowel preparation quality (odds ratio = .418, p = .020, 95% confidence interval [.197, .885]). Low-quality bowel preparation can lead to incomplete readings of the colon or cancelling a colonoscopy. This intervention demonstrates that evidence-based patient navigator programs are effective in increasing screening among a hard-to-reach population and improving bowel preparation quality for males.
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Colonoscopía/estadística & datos numéricos , Neoplasias Colorrectales/prevención & control , Tamizaje Masivo/estadística & datos numéricos , Aceptación de la Atención de Salud/estadística & datos numéricos , Navegación de Pacientes/organización & administración , Neoplasias Colorrectales/diagnóstico , Detección Precoz del Cáncer/estadística & datos numéricos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Educación del Paciente como Asunto , Proyectos de InvestigaciónRESUMEN
Core promoters are crucial for gene regulation, providing blueprints for the assembly of transcriptional machinery at transcription start sites (TSSs). Empirically, TSSs define the coordinates of core promoters and other regulatory sequences. Thus, experimental TSS identification provides an essential step in the characterization of promoters and their features. Here, we describe the application of CAGE (cap analysis of gene expression) to identify genome-wide TSSs used in root and shoot tissues of two maize (Zea mays) inbred lines (B73 and Mo17). Our studies indicate that most TSS clusters are sharp in maize, similar to mice, but distinct from Arabidopsis thaliana, Drosophila melanogaster, or zebra fish, in which a majority of genes have broad-shaped TSS clusters. We established that â¼38% of maize promoters are characterized by a broader TATA-motif consensus, and this motif is significantly enriched in genes with sharp TSSs. A noteworthy plasticity in TSS usage between tissues and inbreds was uncovered, with â¼1500 genes showing significantly different dominant TSSs, sometimes affecting protein sequence by providing alternate translation initiation codons. We experimentally characterized instances in which this differential TSS utilization results in protein isoforms with additional domains or targeted to distinct subcellular compartments. These results provide important insights into TSS selection and gene expression in an agronomically important crop.
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Regulación de la Expresión Génica de las Plantas , Genoma de Planta/genética , Regiones Promotoras Genéticas/genética , Sitio de Iniciación de la Transcripción , Zea mays/genética , Biblioteca de Genes , Genotipo , Motivos de Nucleótidos , Raíces de Plantas/citología , Raíces de Plantas/genética , Brotes de la Planta/citología , Brotes de la Planta/genética , Análisis de Secuencia de ARN , Zea mays/citologíaRESUMEN
Lignin is an essential polymer in vascular plants that plays key structural roles in vessels and fibers. Lignification is induced by external inputs such as wounding, but the molecular mechanisms that link this stress to lignification remain largely unknown. In this work, we provide evidence that three maize (Zea mays) lignin repressors, MYB11, MYB31, and MYB42, participate in wound-induced lignification by interacting with ZML2, a protein belonging to the TIFY family. We determined that the three R2R3-MYB factors and ZML2 bind in vivo to AC-rich and GAT(A/C) cis-elements, respectively, present in a set of lignin genes. In particular, we show that MYB11 and ZML2 bind simultaneously to the AC-rich and GAT(A/C) cis-elements present in the promoter of the caffeic acid O-methyl transferase (comt) gene. We show that, like the R2R3-MYB factors, ZML2 also acts as a transcriptional repressor. We found that upon wounding and methyl jasmonate treatments, MYB11 and ZML2 proteins are degraded and comt transcription is induced. Based on these results, we propose a molecular regulatory mechanism involving a MYB/ZML complex in which wound-induced lignification can be achieved by the derepression of a set of lignin genes.
Asunto(s)
Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Lignina/genética , Zea mays/genética , Acetatos/farmacología , Secuencias de Aminoácidos , Secuencia de Bases , Inmunoprecipitación de Cromatina , Ciclopentanos/farmacología , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Lignina/metabolismo , Modelos Biológicos , Datos de Secuencia Molecular , Oxilipinas/farmacología , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regiones Promotoras Genéticas/genética , Unión Proteica/efectos de los fármacos , Proteolisis/efectos de los fármacos , Zea mays/efectos de los fármacosRESUMEN
Adaptive collision avoidance behaviours require accurate detection of complex spatiotemporal properties of an object approaching in an animal's natural, three-dimensional environment. Within the locust, the lobula giant movement detector and its postsynaptic partner, the descending contralateral movement detector (DCMD), respond robustly to images that emulate an approaching two-dimensional object and exhibit firing rate modulation correlated with changes in object trajectory. It is not known how this pathway responds to visual expansion of a three-dimensional object or an approaching object that changes velocity, both of which represent natural stimuli. We compared DCMD responses with images that emulate the approach of a sphere with those elicited by a two-dimensional disc. A sphere evoked later peak firing and decreased sensitivity to the ratio of the half size of the object to the approach velocity, resulting in an increased threshold subtense angle required to generate peak firing. We also presented locusts with an approaching sphere that decreased or increased in velocity. A velocity decrease resulted in transition-associated peak firing followed by a firing rate increase that resembled the response to a constant, slower velocity. A velocity increase resulted in an earlier increase in the firing rate that was more pronounced with an earlier transition. These results further demonstrate that this pathway can provide motor circuits for behaviour with salient information about complex stimulus dynamics.