Direct Visualization of Single Nuclear Pore Complex Proteins Using Genetically-Encoded Probes for DNA-PAINT

The nuclear pore complex (NPC) is one of the largest and most complex protein assemblies in the cell and, among other functions, serves as the gatekeeper of nucleocytoplasmic transport. Unraveling its molecular architecture and functioning has been an active research topic for decades with recent cryogenic electron microscopy and super-resolution studies advancing our understanding of the architecture of the NPC complex. However, the specific and direct visualization of single copies of NPC proteins is thus far elusive. Herein, we combine genetically-encoded self-labeling enzymes such as SNAP-tag and HaloTag with DNA-PAINT microscopy. We resolve single copies of nucleoporins in the human Y-complex in three dimensions with a precision of circa 3 nm, enabling studies of multicomponent complexes on the level of single proteins in cells using optical fluorescence microscopy

STIM2 regulates PKA-dependent phosphorylation and trafficking of AMPARs

STIMs (STIM1 and STIM2 in mammals) are transmembrane proteins that reside in the endoplasmic reticulum (ER) and regulate store-operated Ca2+ entry (SOCE). The function of STIMs in the brain is only beginning to be explored, and the relevance of SOCE in nerve cells is being debated. Here we identify STIM2 as a central organizer of excitatory synapses. STIM2, but not its paralogue STIM1, influences the formation of dendritic spines and shapes basal synaptic transmission in excitatory neurons. We further demonstrate that STIM2 is essential for cAMP/PKA-dependent phosphorylation of the AMPA receptor (AMPAR) subunit GluA1. cAMP triggers rapid migration of STIM2 to ER–plasma membrane (PM) contact sites, enhances recruitment of GluA1 to these ER-PM junctions, and promotes localization of STIM2 in dendritic spines. Both biochemical and imaging data suggest that STIM2 regulates GluA1 phosphorylation by coupling PKA to the AMPAR in a SOCE-independent manner. Consistent with a central role of STIM2 in regulating AMPAR phosphorylation, STIM2 promotes cAMP-dependent surface delivery of GluA1 through combined effects on exocytosis and endocytosis. Collectively our results point to a unique mechanism of synaptic plasticity driven by dynamic assembly of a STIM2 signaling complex at ER-PM contact sites

Adoption of intercropping among smallholder rubber farmers in Xishuangbanna, China

While the expansion of rubber plantations in the mountainous areas of Southern China has contributed to poverty reduction among the indigenous ethnic groups, the concerns about the associated negatives on ecosystems and sustainable development of livelihoods is rising. One of the measures suggested to reduce environmental and economic risks is rubber intercropping. This study uses cross-section data of some 600 rubber farmers in Xishuangbanna, as a basis to develop four empirical models in order to analyse adoption of intercropping at farm and at plot level. Our study shows that only a small proportion of rubber farmers have adopted intercropping, with tea being the most frequently adopted intercrop. However, we also find that intercropping is an important source of income for the household in the lower income category. Intercropping adoption is affected by ethnicity, household wealth and family labour. The choice of intercrops depends on the nature of rubber plots, the age of rubber trees and geography. This study contributes to a better understanding of the transformation path from rubber monoculture to more diversified, rubber-based agroforestry systems and thus can provide important information for agricultural extension services charged with the promotion of sustainable rubber-based livelihood systems in the Mekong area.German Federal Ministry of Education and Research; FKZ [01LL0919A]; National Natural Science Foundation of China [71333013]SCI(E)SSCIARTICLE3223-2371

Integrating nitrogen fixing structures into above- and belowground functional trait spectra in soy (Glycine max)

This is an accepted manuscript of an article published in Plant and Soil.Aims Phenotypic trait variation across environmental gradients and through plant ontogeny is critical in driving ecological processes, especially in agroecosystems where single genotypes exist in high abundances. While variability in root traits plays a key role in belowground processes, few studies have identified the presence of an intraspecific “Root Economics Spectrum” (RES) within domesticated plants. Furthermore, little is known regarding if an intraspecific RES changes through plant ontogeny, and how trophic interactions – namely root nodulation – relate to above- or belowground trait spectra. Methods We evaluated covariation among 12 root, nodule, leaf, and stem traits in 134 plants of a single genotype of soy (Glycine max). Variation in these traits was assessed across five managed environmental conditions, and three plant ontogenetic stages. Results Root traits covaried along an intraspecific RES that represents a trade-off between resource acquisition and resource conservation. Variation along the RES was closely coordinated with hydraulic traits, but was orthogonal to nodule and leaf economics traits. Trait relationships varied strongly across managed environmental conditions and plant developmental stages. Conclusions Our results indicate the presence of an intraspecific RES in soy that is independent of root nodule investment. Patterns of phenotypic variation in below and aboveground soy traits demonstrate multivariate trait syndromes vary across environmental gradients and are dynamic through plant ontogeny.This research was undertaken in part based on funding from the Canada Research Chairs program and a Natural Sciences and Engineering Research Council of Canada Discovery Grant to Marney E. Isaac, as well as a graduate research assistantship granted to Fallon Hayes courtesy of the Department of Physical and Environmental Sciences, University of Toronto Scarborough

Intraspecific variation in soy across the leaf economics spectrum

Background and Aims: Intraspecific trait variation (ITV) is an important dimension of plant ecological diversity, particularly in agroecosystems, where phenotypic ITV (within crop genotypes) is an important correlate of key agroecosystem processes including yield. There are few studies that have evaluated whether plants of the same genotype vary along well-defined axes of biological variation, such as the leaf economics spectrum (LES). There is even less information disentangling environmental and ontogenetic determinants of crop ITV along an intraspecific LES, and whether or not a plant's position along an intraspecific LES is correlated with reproductive output. Methods: We sought to capture the extent of phenotypic ITV within a single cultivar of soy (Glycine max) - the world's most commonly cultivated legume - using a data set of nine leaf traits measured on 402 leaves, sampled from 134 plants in both agroforestry and monoculture management systems, across three distinct whole-plant ontogenetic stages (while holding leaf age and canopy position stable). Key Results: Leaf traits covaried strongly along an intraspecific LES, in patterns that were largely statistically indistinguishable from the 'universal LES' observed across non-domesticated plants. Whole-plant ontogenetic stage explained the highest proportion of phenotypic ITV in LES traits, with plants progressively expressing more 'resource-conservative' LES syndromes throughout development. Within ontogenetic stages, leaf traits differed systematically across management systems, with plants growing in monoculture expressing more 'resource-conservative' trait syndromes: trends largely owing to an approximately ≥50% increases in leaf mass per area (LMA) in high-light monoculture vs. shaded agroforestry systems. Certain traits, particularly LMA, leaf area and maximum photosynthetic rates, correlated closely with plant-level reproductive output. Conclusions: Phenotypic ITV in soy is governed by constraints in trait trade-offs along an intraspecific LES, which in turn (1) underpins plant responses to managed environmental gradients, and (2) reflects shifts in plant functional biology and resource allocation that occur throughout whole-plant ontogeny

Functional connectivity of the pedunculopontine nucleus and surrounding region in parkinson’s disease

Deep brain stimulation of the pedunculopontine nucleus and surrounding region (PPNR) is a novel treatment strategy for gait freezing in Parkinson’s Disease (PD). However, clinical results have been variable, in part because of the paucity of functional information that might help guide selection of the optimal surgical target. In this study, we use simultaneous magnetoencephalography and local field recordings from the PPNR in seven PD patients, to characterise functional connectivity with distant brain areas at rest. The PPNR was preferentially coupled to brainstem and cingulate regions in the alpha frequency (8-12Hz) band and to the medial motor strip and neighbouring areas in the beta (18-33Hz) band. The distribution of coupling also depended on the vertical distance of the electrode from the pontomesencephalic line: most effects being greatest in the middle PPNR, which may correspond to the caudal pars dissipata of the pedunculopontine nucleus. These observations confirm the crucial position of the PPNR as a functional node between cortical areas such as the cingulate/ medial motor strip and other brainstem nuclei, particularly in the dorsal pons. In particular they suggest a special role for the middle PPNR as this has the greatest functional connectivity with other brain regions

Pedunculopontine nucleus deep brain stimulation in Parkinson's disease: A clinical review

Item does not contain fulltextPedunculopontine nucleus region deep brain stimulation (DBS) is a promising but experimental therapy for axial motor deficits in Parkinson's disease (PD), particularly gait freezing and falls. Here, we summarise the clinical application and outcomes reported during the past 10 years. The published dataset is limited, comprising fewer than 100 cases. Furthermore, there is great variability in clinical methodology between and within surgical centers. The most common indication has been severe medication refractory gait freezing (often associated with postural instability). Some patients received lone pedunculopontine nucleus DBS (unilateral or bilateral) and some received costimulation of the subthalamic nucleus or internal pallidum. Both rostral and caudal pedunculopontine nucleus subregions have been targeted. However, the spread of stimulation and variance in targeting means that neighboring brain stem regions may be implicated in any response. Low stimulation frequencies are typically employed (20-80 Hertz). The fluctuating nature of gait freezing can confound programming and outcome assessments. Although firm conclusions cannot be drawn on therapeutic efficacy, the literature suggests that medication refractory gait freezing and falls can improve. The impact on postural instability is unclear. Most groups report a lack of benefit on gait or limb akinesia or dopaminergic medication requirements. The key question is whether pedunculopontine nucleus DBS can improve quality of life in PD. So far, the evidence supporting such an effect is minimal. Development of pedunculopontine nucleus DBS to become a reliable, established therapy would likely require a collaborative effort between experienced centres to clarify biomarkers predictive of response and the optimal clinical methodology. (c) 2017 International Parkinson and Movement Disorder Society

Pedunculopontine nucleus deep brain stimulation in Parkinson's disease: A clinical review

Pedunculopontine nucleus region deep brain stimulation (DBS) is a promising but experimental therapy for axial motor deficits in Parkinson's disease (PD), particularly gait freezing and falls. Here, we summarise the clinical application and outcomes reported during the past 10 years. The published dataset is limited, comprising fewer than 100 cases. Furthermore, there is great variability in clinical methodology between and within surgical centers. The most common indication has been severe medication refractory gait freezing (often associated with postural instability). Some patients received lone pedunculopontine nucleus DBS (unilateral or bilateral) and some received costimulation of the subthalamic nucleus or internal pallidum. Both rostral and caudal pedunculopontine nucleus subregions have been targeted. However, the spread of stimulation and variance in targeting means that neighboring brain stem regions may be implicated in any response. Low stimulation frequencies are typically employed (20-80 Hertz). The fluctuating nature of gait freezing can confound programming and outcome assessments. Although firm conclusions cannot be drawn on therapeutic efficacy, the literature suggests that medication refractory gait freezing and falls can improve. The impact on postural instability is unclear. Most groups report a lack of benefit on gait or limb akinesia or dopaminergic medication requirements. The key question is whether pedunculopontine nucleus DBS can improve quality of life in PD. So far, the evidence supporting such an effect is minimal. Development of pedunculopontine nucleus DBS to become a reliable, established therapy would likely require a collaborative effort between experienced centres to clarify biomarkers predictive of response and the optimal clinical methodology. © 2017 International Parkinson and Movement Disorder Society