1,211 research outputs found

    Super-resolution imaging and estimation of protein copy numbers at single synapses with DNA-PAINT

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    In the brain, the strength of each individual synapse is defined by the complement of proteins present or the "local proteome." Activity-dependent changes in synaptic strength are the result of changes in this local proteome and posttranslational protein modifications. Although most synaptic proteins have been identified, we still know little about protein copy numbers in individual synapses and variations between synapses. We use DNA-point accumulation for imaging in nanoscale topography as a single-molecule super-resolution imaging technique to visualize and quantify protein copy numbers in single synapses. The imaging technique provides near-molecular spatial resolution, is unaffected by photobleaching, enables imaging of large field of views, and provides quantitative molecular information. We demonstrate these benefits by accessing copy numbers of surface AMPA-type receptors at single synapses of rat hippocampal neurons along dendritic segments

    Differential regulation of local mRNA dynamics and translation following long-term potentiation and depression

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    Decades of work have demonstrated that messenger RNAs (mRNAs) are localized and translated within neuronal dendrites and axons to provide proteins for remodeling and maintaining growth cones or synapses. It remains unknown, however, whether specific forms of plasticity differentially regulate the dynamics and translation of individual mRNA species. To address this, we targeted three individual synaptically localized mRNAs, CamkIIa, β-actin, Psd95, and used molecular beacons to track endogenous mRNA movements. We used reporters and CRISPR/Cas9 gene editing to track mRNA translation in cultured neurons. We found alterations in mRNA dynamic properties occurred during two forms of synaptic plasticity, long-term potentiation (cLTP) and depression (mGluR-LTD). Changes in mRNA dynamics following either form of plasticity resulted in an enrichment of mRNA in the vicinity of dendritic spines. Both the reporters and tagging of endogenous proteins revealed the transcript-specific stimulation of protein synthesis following cLTP or mGluR-LTD. As such, the plasticity-induced enrichment of mRNA near synapses could be uncoupled from its translational status. The enrichment of mRNA in the proximity of spines allows for localized signaling pathways to decode plasticity milieus and stimulate a specific translational profile, resulting in a customized remodeling of the synaptic proteome

    Impact of Grazing Management Strategies on Carbon Sequestration in a Semi-Arid Rangeland, USA

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    The effects of 12 years of grazing management strategies on carbon (C) distribution and sequestration were assessed on a semi-arid mixed-grass prairie in Wyoming, USA. Five grazing treatments were evaluated: non-grazed exclosures; continuous, season-long grazing at a light (22 steer-days ha-1) stocking rate; and, rotationally-deferred, short-duration rotation, and continuous, season-long grazing, all three at a heavy stocking rate (59 steer-days ha-1). Non-grazed exclosures exhibited a large buildup of dead plant material (72% of total aboveground plant matter) and forb biomass represented a large component (35%) of the plant community. Stocking rate, but not grazing strategy, changed plant community composition and decreased surface litter. Light grazing decreased forbs and increased cool-season mid-grasses, resulting in a highly diversified plant community and the highest total production of grasses. Heavy grazing increased warm-season grasses at the expense of the cool-season grasses, which decreased total forage production and opportunity for early season grazing. Compared to the exclosures, all grazing treatments resulted in significantly higher levels of C (6000-9000 kg ha-1) in the surface 15 cm of the soil. Higher levels of soil C with grazing are likely the result of faster litter decomposition and recycling, and redistribution of C within the 0-60 cm plant-soil system. Grazing at an appropriate stocking rate had beneficial effects on plant composition, forage production, and soil C sequestration. Without grazing, deterioration of the plant-soil system is indicated

    Registration of ‘Haymaker’ Intermediate Wheatgrass

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    ‘NU-ARS AC2’ crested wheatgrass [Agropyron cristatum (L.) Gaertn.–A. cristatum var. pectinatum (M. Bieb.) Tzvelev] (Reg. no. CV-28, PI 634507) is a broadly adapted, complex composite population produced by allowing selected plants from fairway-type germplasm accessions to randomly intermate. It originates from collections made by Douglas Dewey, USDA-ARS Plant Geneticist, in the former USSR in 1977. It was released in September 2002 by USDA-ARS; Agricultural Research Division, Institute of Agricultural and Natural Resources, University of Nebraska-Lincoln; and the USDA-NRCS. NU-ARS AC2 was tested under the experimental designation NE AC2

    Registration of ‘NU-ARS AC2’ Crested Wheatgrass

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    ‘NU-ARS AC2’ crested wheatgrass [Agropyron cristatum (L.) Gaertn.–A. cristatum var. pectinatum (M. Bieb.) Tzvelev] (Reg. no. CV-28, PI 634507) is a broadly adapted, complex composite population produced by allowing selected plants from fairway-type germplasm accessions to randomly intermate. It originates from collections made by Douglas Dewey, USDA-ARS Plant Geneticist, in the former USSR in 1977. It was released in September 2002 by USDA-ARS; Agricultural Research Division, Institute of Agricultural and Natural Resources, University of Nebraska-Lincoln; and the USDA-NRCS. NU-ARS AC2 was tested under the experimental designation NE AC2

    Association of Pyrethroid Pesticide Exposure With Attention-Deficit/Hyperactivity Disorder in a Nationally Representative Sample of U.S. Children

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    Background Pyrethroid pesticides cause abnormalities in the dopamine system and produce an ADHD phenotype in animal models, with effects accentuated in males versus females. However, data regarding behavioral effects of pyrethroid exposure in children is limited. We examined the association between pyrethroid pesticide exposure and ADHD in a nationally representative sample of US children, and tested whether this association differs by sex. Methods Data are from 8–15 year old participants (N = 687) in the 2001–2002 National Health and Nutrition Examination Survey. Exposure was assessed using concurrent urinary levels of the pyrethroid metabolite 3-phenoxybenzoic acid (3-PBA). ADHD was defined by either meeting Diagnostic and Statistical Manual of Mental Disorders-Fourth Edition criteria on the Diagnostic Interview Schedule for Children (DISC) or caregiver report of a prior diagnosis. ADHD symptom counts were determined via the DISC. Multivariable logistic regression examined the link between pyrethroid exposure and ADHD, and poisson regression investigated the link between exposure and ADHD symptom counts. Results Children with urinary 3-PBA above the limit of detection (LOD) were twice as likely to have ADHD compared with those below the LOD (adjusted odds ratio [aOR] 2.42; 95 % confidence interval [CI] 1.06, 5.57). Hyperactive-impulsive symptoms increased by 50 % for every 10-fold increase in 3-PBA levels (adjusted count ratio 1.50; 95 % CI 1.03, 2.19); effects on inattention were not significant. We observed possible sex-specific effects: pyrethroid biomarkers were associated with increased odds of an ADHD diagnosis and number of ADHD symptoms for boys but not girls. Conclusions We found an association between increasing pyrethroid pesticide exposure and ADHD which may be stronger for hyperactive-impulsive symptoms compared to inattention and in boys compared to girls. Given the growing use of pyrethroid pesticides, these results may be of considerable public health import
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