Volume-regulated Cl- current: contributions of distinct Cl- channel and localized Ca2+ signals.

The swelling-activated chloride current (ICl,swell) is induced when a cell swells and plays a central role in maintaining cell volume in response to osmotic stress. The major contributor of ICl,swell is the volume regulated anion channel (VRAC). LRRC8A (SWELL1) was recently identified as an essential component of VRAC but the mechanisms of VRAC activation are still largely unknown; moreover, other Cl- channels, such as anoctamin 1 (ANO1) were also suggested to contribute to ICl,swell. In this present study, we investigated the roles of LRRC8A and ANO1 in activation of ICl,swell; we also explored the role of intracellular Ca2+ in ICl,swell activation. We used CRISPR/Cas9 gene editing approach, electrophysiology, live fluorescent imaging, selective pharmacology and other approaches to show that both LRRC8A and ANO1 can be activated by cell swelling in HEK293 cells. Yet, both channels contribute biophysically and pharmacologically distinct components to ICl,swell, with LRRC8A being the major component. Cell swelling induced oscillatory Ca2+ transients and these Ca2+ signals were required to activate both, the LRRC8A- and ANO1-dependent components of ICl,swell. Both ICl,swell components required localized rather than global Ca2+ for activation. Interestingly, while intracellular Ca2+ was necessary and sufficient to activate ANO1, it was necessary but not sufficient to activate LRRC8A-mediated currents. Finally, Ca2+ transients linked to the ICl,swell activation were mediated by the GPCR-independent PLC isoforms

Dynamic Changes in Dietary Guideline Adherence and Its Association with All-Cause Mortality among Middle-Aged Chinese: A Longitudinal Study from the China Health and Nutrition Survey

The traditional approach to evaluating dietary quality is based on the achievement of the recommended intakes for each food group, which may overlook the achievement of correct relative proportions between food groups. We propose a “Dietary Non-Adherence Score (DNAS)” to assess the degree of similarity between subjects’ diets and those recommended in the Chinese Dietary Guidelines (CDG). Furthermore, it is important to incorporate the time-dependent nature of dietary quality into mortality prediction. This study investigated the association between long-term changes in adherence to the CDG and all-cause mortality. This study included 4533 participants aged 30–60 from the China Health and Nutrition Survey study with a median follow-up of 6.9 years. Intakes from 10 food groups were collected in 5 survey rounds from 2004 to 2015. We calculated the Euclidean distance between the intake of each food and the CDG-recommended intake, and then summed all the food groups as DNAS. Mortality was assessed in 2015. Latent class trajectory modeling was used to identify three classes of participants with distinct longitudinal trajectories of DNAS during the follow-up period. The Cox proportional hazard model was used to assess the risk of all-cause mortality in the three classes of people. Risk factors for death and confounders for diets were sequentially adjusted in the models. There were 187 deaths overall. Participants in the first class identified had consistently low and decreasing DNAS levels (coefficient = −0.020) over their lifetime, compared with a hazard ratio (HR) of 4.4 (95% confidence interval [CI]: 1.5, 12.7) for participants with consistently high and increasing DNAS levels (coefficient = 0.008). Those with moderate DNAS had an HR of 3.0 (95% CI: 1.1, 8.4). In summary, we find that people with consistently high adherence to CDG-recommended dietary patterns had a significantly lower mortality risk. DNAS is a promising method to assess diet quality

De novo transcriptome analysis of Medicago falcata reveals novel insights about the mechanisms underlying abiotic stress-responsive pathway

BACKGROUND: The entire world is facing a deteriorating environment. Understanding the mechanisms underlying plant responses to external abiotic stresses is important for breeding stress-tolerant crops and herbages. Phytohormones play critical regulatory roles in plants in the response to external and internal cues to regulate growth and development. Medicago falcata is one of the stress-tolerant candidate leguminous species and is able to fix atmospheric nitrogen. This ability allows leguminous plants to grow in nitrogen deficient soils. METHODS: We performed Illumina sequencing of cDNA prepared from abiotic stress treated M. falcata. Sequencedreads were assembled to provide a transcriptome resource. Transcripts were annotated using BLASTsearches against the NCBI non-redundant database and gene ontology definitions were assigned. Acomparison among the three abiotic stress treated samples was carried out. The expression of transcriptswas confirmed with qRT-PCR. RESULTS: We present an abiotic stress-responsive M. falcata transcriptome using next-generation sequencing data from samples grown under standard, dehydration, high salinity, and cold conditions. We combined reads from all samples and de novo assembled 98,515 transcripts to build the M. falcata gene index. A comprehensive analysis of the transcriptome revealed abiotic stress-responsive mechanisms underlying the metabolism and core signalling components of major phytohormones. We identified nod factor signalling pathways during early symbiotic nodulation that are modified by abiotic stresses. Additionally, a global comparison of homology between the M. falcata and M. truncatula transcriptomes, along with five other leguminous species, revealed a high level of global sequence conservation within the family. CONCLUSIONS: M. falcata is shown to be a model candidate for studying abiotic stress-responsive mechanisms in legumes. This global gene expression analysis provides new insights into the biochemical and molecular mechanisms involved in the acclimation to abiotic stresses. Our data provides many gene candidates that might be used for herbage and crop breeding. Additionally, FalcataBase (http://bioinformatics.cau.edu.cn/falcata/) was built for storing these data. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-2019-x) contains supplementary material, which is available to authorized users

Neuropathic Injury-Induced Plasticity of GABAergic System in Peripheral Sensory Ganglia

GABA is a major inhibitory neurotransmitter in the mammalian central nervous system (CNS). Inhibitory GABAA channel circuits in the dorsal spinal cord are the gatekeepers of the nociceptive input from the periphery to the CNS. Weakening of these spinal inhibitory mechanisms is a hallmark of chronic pain. Yet, recent studies have suggested the existence of an earlier GABAergic “gate” within the peripheral sensory ganglia. In this study, we performed systematic investigation of plastic changes of the GABA-related proteins in the dorsal root ganglion (DRG) in the process of neuropathic pain development. We found that chronic constriction injury (CCI) induced general downregulation of most GABAA channel subunits and the GABA-producing enzyme, glutamate decarboxylase, consistent with the weakening of the GABAergic inhibition at the periphery. Strikingly, the α5 GABAA subunit was consistently upregulated. Knock-down of the α5 subunit in vivo moderately alleviated neuropathic hyperalgesia. Our findings suggest that while the development of neuropathic pain is generally accompanied by weakening of the peripheral GABAergic system, the α5 GABAA subunit may have a unique pro-algesic role and, hence, might represent a new therapeutic target

Inflammatory mediator bradykinin increases population of sensory neurons expressing functional T-type Ca2+ channels

T-type Ca2+ channels are important regulators of peripheral sensory neuron excitability. Accordingly, T-type Ca2+ currents are often increased in various pathological pain conditions, such as inflammation or nerve injury. Here we investigated effects of inflammation on functional expression of T-type Ca2+ channels in small-diameter cultured dorsal root ganglion (DRG) neurons. We found that overnight treatment of DRG cultures with a cocktail of inflammatory mediators bradykinin (BK), adenosine triphosphate (ATP), norepinephrine (NE) and prostaglandin E2 (PGE2) strongly increased the population size of the small-diameter neurons displaying low-voltage activated (LVA, T-type) Ca2+ currents while having no effect on the peak LVA current amplitude. When applied individually, BK and ATP also increased the population size of LVA-positive neurons while NE and PGE2 had no effect. The PLC inhibitor U-73122 and B2 receptor antagonist, Hoe-140, both abolished the increase of the population of LVA-positive DRG neurons. Inflammatory treatment did not affect CaV3.2 mRNA or protein levels in DRG cultures. Furthermore, an ubiquitination inhibitor, MG132, did not increase the population of LVA-positive neurons. Our data suggest that inflammatory mediators BK and ATP increase the abundance of LVA-positive DRG neurons in total neuronal population by stimulating the recruitment of a 'reserve pool' of CaV3.2 channels, particularly in neurons that do not display measurable LVA currents under control conditions

Improving maize’s N uptake and N use efficiency by strengthening roots’ absorption capacity when intercropped with legumes

Maize’s nitrogen (N) uptake can be improved through maize-legume intercropping. N uptake mechanisms require further study to better understand how legumes affect root growth and to determine maize’s absorptive capacity in maize-legume intercropping. We conducted a two-year field experiment with two N treatments (zero N (N0) and conventional N (N1)) and three planting patterns (monoculture maize (Zea mays L.) (MM), maize-soybean (Glycine max L. Merr.) strip intercropping (IMS), and maize-peanut (Arachis hypogaea L.) strip intercropping (IMP)). We sought to understand maize’s N uptake mechanisms by investigating root growth and distribution, root uptake capacity, antioxidant enzyme activity, and the antioxidant content in different maize-legume strip intercropping systems. Our results showed that on average, the N uptake of maize was significantly greater by 52.5% in IMS and by 62.4% in IMP than that in MM. The average agronomic efficiency (AE) of maize was increased by 110.5 % in IMS and by 163.4 % in IMP, compared to MM. The apparent recovery efficiency (RE) of maize was increased by 22.3% in IMS. The roots of intercropped maize were extended into soybean and peanut stands underneath the space and even between the inter-rows of legume, resulting in significantly increased root surface area density (RSAD) and total root biomass. The root-bleeding sap intensity of maize was significantly increased by 22.7–49.3% in IMS and 37.9–66.7% in IMP, compared with the MM. The nitrate-N content of maize bleeding sap was significantly greater in IMS and IMP than in MM during the 2018 crop season. The glutathione (GSH) content, superoxide dismutase (SOD), and catalase (CAT) activities in the root significantly increased in IMS and IMP compared to MM. Strip intercropping using legumes increases maize’s aboveground N uptake by promoting root growth and spatial distribution, delaying root senescence, and strengthening root uptake capacity

Baicalin Attenuates Oxygen–Glucose Deprivation/Reoxygenation–Induced Injury by Modulating the BDNF-TrkB/PI3K/Akt and MAPK/Erk1/2 Signaling Axes in Neuron–Astrocyte Cocultures

Background: Baicalin (BCL), a candidate drug for ischemic stroke, has been indicated to protect neurons by promoting brain-derived neurotrophic factor (BDNF). However, the cellular source of BDNF release promoted by baicalin and its detailed protective mechanism after ischemia/reperfusion remains to be studied. The aim of this study was to investigate the neuroprotective mechanisms of baicalin against oxygen–glucose deprivation/reoxygenation (OGD/R) in a neuron–astrocyte coculture system and to explore whether the BDNF-TrkB pathway is involved.Methods and Results: A neuron–astrocyte coculture system was established to elucidate the role of astrocytes in neurons under OGD/R conditions. The results demonstrated that astrocytes became reactive astrocytes and released more BDNF in the coculture system to attenuate neuronal apoptosis and injury after OGD/R. BCL maintained the characteristics of reactive astrocytes and obviously increased the expression of cyclic AMP response element-binding protein (CREB) and the levels of BDNF in the coculture system after OGD/R. To further verify whether BDNF binding to its receptor tyrosine kinase receptor B (TrkB) was required for the neuroprotective effect of baicalin, we examined the effect of ANA-12, an antagonist of TrkB, on NA system injury, including oxidative stress, inflammation, and apoptosis induced by OGD/R. The results showed that treatment of NA systems with ANA-12 significantly attenuated the neuroprotection of BCL. The phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) and mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) pathways are two important downstream cascades of signaling pathways activated by BDNF binding to TrkB. We investigated the expressions of TrkB, PI3K, Akt, MAPK, and ERK. The results demonstrated that baicalin significantly increased the expressions of TrkB, PI3K/AKT, and MAPK/ERK.Conclusion: The neuroprotective effects of baicalin against oxidative stress, inflammation, and apoptosis were improved by astrocytes, mainly mediated by increasing the release of BDNF and its associated receptor TrkB and downstream signaling regulators PI3K/Akt and MAPK/ERK1/2

Local GABAergic signaling within sensory ganglia controls peripheral nociceptive transmission

The integration of somatosensory information is generally assumed to be a function of the central nervous system (CNS). Here we describe fully functional GABAergic communication within rodent peripheral sensory ganglia and show that it can modulate transmission of pain-related signals from the peripheral sensory nerves to the CNS. We found that sensory neurons express major proteins necessary for GABA synthesis and release and that sensory neurons released GABA in response to depolarization. In vivo focal infusion of GABA or GABA reuptake inhibitor to sensory ganglia dramatically reduced acute peripherally induced nociception and alleviated neuropathic and inflammatory pain. In addition, focal application of GABA receptor antagonists to sensory ganglia triggered or exacerbated peripherally induced nociception. We also demonstrated that chemogenetic or optogenetic depolarization of GABAergic dorsal root ganglion neurons in vivo reduced acute and chronic peripherally induced nociception. Mechanistically, GABA depolarized the majority of sensory neuron somata, yet produced a net inhibitory effect on the nociceptive transmission due to the filtering effect at nociceptive fiber T-junctions. Our findings indicate that peripheral somatosensory ganglia represent a hitherto underappreciated site of somatosensory signal integration and offer a potential target for therapeutic intervention