An open-access volume electron microscopy atlas of whole cells and tissues.

Understanding cellular architecture is essential for understanding biology. Electron microscopy (EM) uniquely visualizes cellular structures with nanometre resolution. However, traditional methods, such as thin-section EM or EM tomography, have limitations in that they visualize only a single slice or a relatively small volume of the cell, respectively. Focused ion beam-scanning electron microscopy (FIB-SEM) has demonstrated the ability to image small volumes of cellular samples with 4-nm isotropic voxels1. Owing to advances in the precision and stability of FIB milling, together with enhanced signal detection and faster SEM scanning, we have increased the volume that can be imaged with 4-nm voxels by two orders of magnitude. Here we present a volume EM atlas at such resolution comprising ten three-dimensional datasets for whole cells and tissues, including cancer cells, immune cells, mouse pancreatic islets and Drosophila neural tissues. These open access data (via OpenOrganelle2) represent the foundation of a field of high-resolution whole-cell volume EM and subsequent analyses, and we invite researchers to explore this atlas and pose questions.

Phosphatase, Mg2+/Mn2+ dependent 1B regulates the hematopoietic stem cells homeostasis via the Wnt/ß-catenin signaling.

Hematopoietic stem cells (HSC) are primarily dormant in a cell-cycle quiescence state to preserve their self-renewal capacity and long-term maintenance. How HSC maintain the balance between activation and quiescence remains largely unknown. Herein, we found that phosphatase, Mg2+/Mn2+ dependent 1B (Ppm1b) is required for the expansion of phenotypic HSC in vitro. By using a conditional knockout mouse model in which Ppm1b was specifically depleted in hematopoietic cells, we demonstrated that loss of Ppm1b impaired the HSC homeostasis and hematopoietic reconstitution. Ppm1b deficiency mice also exhibited B-cell leukocytopenia, which is due to the compromised commitment and proliferation of B-biased lymphoid progenitor cells from common lymphoid progenitors. With the aid of a small molecular inhibitor, we confirmed the roles of Ppm1b in adult hematopoiesis that phenocopied the effects with loss of Ppm1b. Furthermore, transcriptome profiling of Ppm1b-deficient HSC revealed the disruptive quiescence of HSC. Mechanistically, Ppm1b interacted with ß-catenin and mediated its dephosphorylation. Loss of Ppm1b led to the decrease in the active ß-catenin (non-phosphorylated) that interrupted the Wnt/ß-catenin signaling in HSC, which consequently suppressed HSC expansion. Together, our study identified an indispensable role for Ppm1b in regulating HSC homeostasis via the Wnt/ß-catenin pathway.

The impact of crowding stress on growth and intestinal integrity in largemouth bass (Micropterus salmoides): Insights into ER stress, autophagy and apoptosis.

Crowding stress is a common abiotic stressor in intensive aquaculture that negatively impacts fish species, leading to growth depression. This study primarily explored the effects of crowding stress on the growth and intestinal integrity of largemouth bass (Micropterus salmoides). A 10-week feeding experiment was conducted with two groups: a control group (0.55 kg/m³) and a crowding stress group (1.10 kg/m³). The results showed that crowding stress significantly impaired fish growth and compromised intestinal integrity. To further elucidate the underlying mechanisms, we investigated the effects of crowding stress on endoplasmic reticulum (ER) stress, autophagy and apoptosis. Our findings revealed an increased proportion of terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL)-positive cells and ER swelling in the intestines of stressed fish, along with a higher number of autophagosomes. Furthermore, there was a significant upregulation in the mRNA expression of genes related to ER stress, autophagy and apoptosis, including GRP78, eIF2α, IRE1, atg3, LC3-2, ulk1α, P62, and caspase-8. In conclusion, crowding stress negatively affects fish growth and intestinal integrity, potentially through the induction of apoptosis and autophagy associated with ER stress-mediated unfolded protein response (UPR). These findings provided critical insights into how intensive aquaculture disrupts intestinal integrity and inhibits growth in fish, offering a valuable reference for future research aimed at enhancing stress resistance in fish under intensive aquaculture conditions.

All-hydrocarbon stapling enables improvement of antimicrobial activity and proteolytic stability of peptide Figainin 2.

Figainin 2 is a cationic, hydrophobic, α-helical host-defense peptide with 28 residues, which was isolated from the skin secretions of the Chaco tree frog. It shows potent inhibitory activity against both Gram-negative and Gram-positive pathogens and has garnered considerable interest in developing novel classes of natural antibacterial agents. However, as a linear peptide, conformational flexibility and poor proteolytic stability hindered its development as antibacterial agent. To alleviate its susceptibility to proteolytic degradation and improve its antibacterial activity, a series of hydrocarbon-stable analogs of Figainin 2 were synthesized and evaluated for their secondary structure, protease stability, antimicrobial, and hemolytic activities. Among them, F2-12 showed significant improvement in protease resistance and antimicrobial activity compared to that of the template peptide. This study provides a promising strategy for the development of antimicrobial drugs.

Chemerin promotes invasion of oral squamous cell carcinoma by stimulating IL-6 and TNF-α production via STAT3 activation.

AIMS: Elevated levels of adipokine chemerin have been identified in oral squamous cell carcinoma (OSCC) and found to be associated with metastasis to the cervical lymph nodes. The underlying mechanism through which chemerin affects OSCC progression is unclear. The aims of this study were firstly to determine chemerin levels and cytokine concentrations in serum from patients with OSCC and in OSCC cell cultures, and secondly to observe chemerin effects on OSCC cell cytokine secretion, migration, and invasion in vitro. METHODS: Serum samples were collected from 20 patients diagnosed with OSCC, including groups with (LN+) and without (LN-) cervical lymph node metastasis. A Luminex liquid suspension assay was used to quantify serum concentrations of 27 types of cytokines. Correlations between chemerin and cytokines (i.e., IL-6, IL-15, GM-CSF, RANTES, TNF-α, and VEGF) were analyzed. ELISAs (enzyme-linked immunosorbent assays) were used to determine concentrations of chemerin and selected cytokines in serum and in supernatants of OSCC cell cultures (SCC9 and SCC25 cell lines). OSCC cells were stimulated with human recombinant chemerin, STAT3 inhibitor, or IL-6 together with TNF-α neutralizing antibodies. Phosphorylated STAT3 protein levels were measured with western blot analysis. OSCC cell migration and invasion were investigated with Transwell assays. RESULTS: Compared to the LN- group, OSCC patients with cervical lymph node metastasis had higher levels of IL-6 (P = 0.006), IL-15 (P = 0.020), GM-CSF (P = 0.036), RANTES (P = 0.032), TNF-α (P = 0.005), VEGF (P = 0.006), and chemerin (P = 0.001). Patients' serum chemerin levels correlated directly with IL-6, GM-CSF, TNF-α, and VEGF levels in OSCC patients. Exogenous recombinant chemerin treatment promoted secretion of IL-6 and TNF-α via activation of STAT3 in OSCC cells. Chemerin induced OSCC-cell migration and invasion, and these effects were reduced by IL-6 and TNF-α neutralizing antibodies. CONCLUSION: Our findings indicate that chemerin may play a role in advancing OSCC progression by increasing production of IL-6 and TNF-α, perhaps via a mechanism involving STAT3 signaling.

Associations between social support and proactive health behaviours among Chinese adolescents: the mediating role of self-efficacy and peer relationships.

BACKGROUND: Proactive health behaviours are crucial for enhancing adolescent health. However, there is limited evidence on the potential pathways through which social support influences adolescents' proactive health behaviours. This cross-sectional study aimed to examine the relationships between social support, self-efficacy, peer relationships and proactive health behaviours in Chinese adolescents. METHODS: From October to December 2023, we recruited 6075 adolescents from Shandong Province, China. They completed self-report questionnaires on social support, self-efficacy, peer relationships and proactive health behaviours. RESULTS: Linear regression analysis indicated that social support was positively associated with proactive health behaviours among adolescents (ß = 0.571, 95% CI = 0.542, 0.600). Further mediation analyses revealed that self-efficacy (ß = 0.085, 95% CI = 0.069,0.101) and peer relationships (ß = 0.156, 95% CI = 0.136,0.177) mediated this relationship. CONCLUSIONS: Increased social support was associated with better proactive health behaviours in Chinese adolescents. Additionally, higher self-efficacy and positive peer relationships enhanced this association. Our findings emphasised the significance of providing supportive environments at home and at school to promote proactive health behaviours in adolescents.

Association between loneliness and mild cognitive impairment in older adults: a meta-analysis of longitudinal studies.

OBJECTIVES: Prior studies reporting the effects of loneliness on mild impairment cognitive (MCI) have generated inconsistent results. This meta-analysis aimed to investigate the longitudinal association between loneliness and risk of MCI among community-dwelling middle-aged and older adults. METHOD: Five electronic databases were searched from inception to 9 May 2023. Eligible studies examined the longitudinal association between loneliness and cognitive outcomes, including incident MCI, cognitive impairment, and cognitive decline. Odds ratios (OR) and 95% confidence intervals (CIs) were calculated using random-effects or fixed-effects meta-analysis. Sensitivity analysis and subgroup analysis were conducted. Publication bias was examined using Egger's and Begg tests. RESULTS: Eight studies were included. Among the 45,032 participants, 10,570 were diagnosed with MCI/cognitive decline. Loneliness was positively associated with an increased risk of MCI (overall OR = 1.14; 95% CI = 1.05, 1.23), with moderate heterogeneity (I2 = 44.2%). Sensitivity analysis have minimal influence on the aforementioned pooled effect. Subgroup analyses indicated stronger associations in studies which employed incident MCI as cognitive outcome (OR = 2.55, 95%CI = 1.31, 1.83), were conducted in non-Asia countries (OR = 1.52, 95%CI = 0.95, 1.20), and reported no depression adjustment (OR = 1.51, 95%CI = 1.04, 1.25). The association between loneliness and MCI was stronger among males compare to females. The Egger test and Begg test showed no evidence of significant publication bias (p = .493; p = .474). CONCLUSION: The findings indicated that loneliness was associated with an increased risk of MCI. Future longitudinal studies should evaluate potential cases of MCI through comprehensive clinical assessments by practitioners to draw robust findings on the association of loneliness with MCI.

Loneliness was associated with an increased risk of MCI in older adults.The association between loneliness and MCI was stronger in the males compared to the females.Future studies are warranted to assess loneliness by validated scales and clinical assessments of MCI.

Spatio-temporal dynamics of phytoplankton community structure in the coastal waters of the Southern Beibu Gulf.

This study was conducted during October 2021 (autumn) and April 2022 (spring) to explore the phytoplankton community structure, their distribution characteristics, and the influence of environmental factors in the coastal waters of the Southern Beibu Gulf. The 15 sampling sites were grouped based on the difference in offshore distance to analyze the temporal and spatial differences in community structure and environmental driving in the investigated sea area of the coastal waters of the Southern Beibu Gulf. Permutational multivariate analysis of variance was conducted on the sample data in time and space, revealing that there is no significant difference in space (p > 0.05), but there is significant difference in time (p < 0.05). Notably, water pressure, pH, chemical oxygen demand, nitrite, and labile phosphate were higher in autumn, while total ammonia nitrogen, dissolved oxygen, and suspended solids were significantly higher in spring. Additionally, the study identified 87 phytoplankton species belonging to 6 phyla, dominating by Bacillariophyta, followed by Dinophyta and Cyanophyta. The phytoplankton density, Shannon Weiner's diversity index (H'), Pielou's evenness index (J), and Margalef's richness index (D) ranged from 84.88 to 4675.33 cells L-1, 0.56 to 2.58, 0.26 to 0.89, and 1.21 to 3.64, respectively. Permutational multivariate analysis of variance showed non-significant spatial differences in phytoplankton composition (p > 0.05) but seasonal differences (p < 0.05). Furthermore, canonical correspondence analysis (CCA) identified pH, dissolved oxygen, suspended solids, chemical oxygen demand, nitrite, and labile phosphate as key environmental factors influencing the phytoplankton community structure (p < 0.05). In this study, the dynamic changes of phytoplankton community structure and environmental factors in the southern coastal waters of Beibu Gulf were analyzed in detail from two aspects of time and space. The key environmental factors to protect the ecological environment in the southern coastal area of Beibu Gulf were found out. It provides a reference method and theoretical basis for the management and protection of Beibu Gulf and other tropical marine environment.

Presynaptic Mitochondrial Volume and Packing Density Scale with Presynaptic Power Demand.

Stable neural function requires an energy supply that can meet the intense episodic power demands of neuronal activity. Neurons have presumably optimized the volume of their bioenergetic machinery to ensure these power demands are met, but the relationship between presynaptic power demands and the volume available to the bioenergetic machinery has never been quantified. Here, we estimated the power demands of six motor nerve terminals in female Drosophila larvae through direct measurements of neurotransmitter release and Ca2+ entry, and via theoretical estimates of Na+ entry and power demands at rest. Electron microscopy revealed that terminals with the highest power demands contained the greatest volume of mitochondria, indicating that mitochondria are allocated according to presynaptic power demands. In addition, terminals with the greatest power demand-to-volume ratio (∼66 nmol·min-1·µl-1) harbor the largest mitochondria packed at the greatest density. If we assume sequential and complete oxidation of glucose by glycolysis and oxidative phosphorylation, then these mitochondria are required to produce ATP at a rate of 52 nmol·min-1·µl-1 at rest, rising to 963 during activity. Glycolysis would contribute ATP at 0.24 nmol·min-1·µl-1 of cytosol at rest, rising to 4.36 during activity. These data provide a quantitative framework for presynaptic bioenergetics in situ, and reveal that, beyond an immediate capacity to accelerate ATP output from glycolysis and oxidative phosphorylation, over longer time periods presynaptic terminals optimize mitochondrial volume and density to meet power demand.SIGNIFICANCE STATEMENT The remarkable energy demands of the brain are supported by the complete oxidation of its fuel but debate continues regarding a division of labor between glycolysis and oxidative phosphorylation across different cell types. Here, we exploit the neuromuscular synapse, a model for studying neurophysiology, to elucidate fundamental aspects of neuronal energy metabolism that ultimately constrain rates of neural processing. We quantified energy production rates required to sustain activity at individual nerve terminals and compared these with the volume capable of oxidative phosphorylation (mitochondria) and glycolysis (cytosol). We find strong support for oxidative phosphorylation playing a primary role in presynaptic terminals and provide the first in vivo estimates of energy production rates per unit volume of presynaptic mitochondria and cytosol.

Publisher Correction: Gas cluster ion beam SEM for imaging of large tissue samples with 10 nm isotropic resolution.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Gas cluster ion beam SEM for imaging of large tissue samples with 10 nm isotropic resolution.

We demonstrate gas cluster ion beam scanning electron microscopy (SEM), in which wide-area ion milling is performed on a series of thick tissue sections. This three-dimensional electron microscopy technique acquires datasets with <10 nm isotropic resolution of each section, and these can then be stitched together to span the sectioned volume. Incorporating gas cluster ion beam SEM into existing single-beam and multibeam SEM workflows should be straightforward, increasing reliability while improving z resolution by a factor of three or more.

Statistical methods for exploring spontaneous adverse event reporting databases for drug-host factor interactions.

BACKGROUND: Drug toxicity does not affect patients equally; the toxicity may only exert in patients who possess certain attributes of susceptibility to specific drug properties (i.e., drug-host interaction). This concept is crucial for personalized drug safety but remains under-studied, primarily due to methodological challenges and limited data availability. By monitoring a large volume of adverse event reports in the postmarket stage, spontaneous adverse event reporting systems provide an unparalleled resource of information for adverse events and could be utilized to explore risk disparities of specific adverse events by age, sex, and other host factors. However, well-formulated statistical methods to formally address such risk disparities are currently lacking. METHODS: In this paper, we present a statistical framework to explore spontaneous adverse event reporting databases for drug-host interactions and detect risk disparities in adverse drug events by various host factors, adapting methods for safety signal detection. We proposed four different methods, including likelihood ratio test, normal approximation test, and two tests using subgroup ratios. We applied our proposed methods to simulated data and Food and Drug Administration (FDA) Adverse Event Reporting Systems (FAERS) and explored sex-/age-disparities in reported liver events associated with specific drug classes. RESULTS: The simulation result demonstrates that two tests (likelihood ratio, normal approximation) can detect disparities in adverse drug events associated with host factors while controlling the family wise error rate. Application to real data on drug liver toxicity shows that the proposed method can be used to detect drugs with unusually high level of disparity regarding a host factor (sex or age) for liver toxicity or to determine whether an adverse event demonstrates a significant unbalance regarding the host factor relative to other events for the drug. CONCLUSION: Though spontaneous adverse event reporting databases require careful data processing and inference, the sheer size of the databases with diverse data from different countries provides unique resources for exploring various questions for drug safety that are otherwise impossible to address. Our proposed methods can be used to facilitate future investigation on drug-host interactions in drug toxicity using a large number of reported adverse events.

IRDC-Net: An Inception Network with a Residual Module and Dilated Convolution for Sign Language Recognition Based on Surface Electromyography.

Deaf and hearing-impaired people always face communication barriers. Non-invasive surface electromyography (sEMG) sensor-based sign language recognition (SLR) technology can help them to better integrate into social life. Since the traditional tandem convolutional neural network (CNN) structure used in most CNN-based studies inadequately captures the features of the input data, we propose a novel inception architecture with a residual module and dilated convolution (IRDC-net) to enlarge the receptive fields and enrich the feature maps, applying it to SLR tasks for the first time. This work first transformed the time domain signal into a time-frequency domain using discrete Fourier transformation. Second, an IRDC-net was constructed to recognize ten Chinese sign language signs. Third, the tandem CNN networks VGG-net and ResNet-18 were compared with our proposed parallel structure network, IRDC-net. Finally, the public dataset Ninapro DB1 was utilized to verify the generalization performance of the IRDC-net. The results showed that after transforming the time domain sEMG signal into the time-frequency domain, the classification accuracy (acc) increased from 84.29% to 91.70% when using the IRDC-net on our sign language dataset. Furthermore, for the time-frequency information of the public dataset Ninapro DB1, the classification accuracy reached 89.82%; this value is higher than that achieved in other recent studies. As such, our findings contribute to research into SLR tasks and to improving deaf and hearing-impaired people's daily lives.

Cellular antioxidant mechanism of mannan-oligosaccharides involving in enhancing flesh quality in grass carp (Ctenopharyngodon idella).

BACKGROUND: Deterioration of flesh quality has bad effects on consumer satisfaction. Therefore, effects of safe mannan-oligosaccharides (MOS) on flesh quality of grass carp (Ctenopharyngodon idella) muscle were studied. A total of 540 healthy fish (215.85 ± 0.30 g) were randomly divided into six groups and fed six separate diets with graded levels of MOS (0, 200, 400, 600, 800 and 1000 mg kg-1 ) for 60 days. This study aimed at investigating the benefits of dietary MOS on flesh quality (fatty acids, amino acids and physicochemical properties) and the protection mechanism regarding antioxidant status. RESULTS: Optimal MOS could improve tenderness (27.4%), pH (5.5%) while decreasing cooking loss (16.6%) to enhance flesh quality. Meanwhile, optimal MOS improved flavor inosine 5'-monophosphate (IMP) of 11.8%, sweetness and umami-associated amino acid, healthy unsaturated fatty acid (UFA) of 14.9% and n-3 polyunsaturated fatty acids (n-3 PUFAs) especially C20:5n-3 (15.8%) and C22:6n-3 (38.3%). Furthermore, the mechanism that MOS affected pH, tenderness and cooking loss could be partly explained by the reduced lactate, cathepsin and oxidation, respectively. The enhanced flesh quality was also associated with enhanced antioxidant ability concerning improving antioxidant enzymes activities and the corresponding transcriptional levels, which were regulated through NF-E2-related factor 2 (Nrf2) and target of rapamycin (TOR) signaling. Based on pH24h , cooking loss, shear force and DHA (docosahexaenoic acid, C22:6n-3), optimal MOS levels for grass carp were estimated to be 442.75, 539.53, 594.73 and 539.53 mg kg-1 , respectively. CONCLUSION: Dietary MOS is a promising alternative strategy to improve flesh quality of fish muscle. © 2022 Society of Chemical Industry.

MScanFit motor unit number estimation of human anconeus muscle.

INTRODUCTION/AIMS: Motor unit number estimation (MUNE) studies of the anconeus muscle are very limited, although the information they provide is useful for neurophysiological investigations. The objective of this study was to estimate the number of motor units in the anconeus muscle. METHODS: Compound muscle action potential scans of the anconeus muscle were recorded from 11 healthy participants, all of whom were tested on two occasions. MUNE was determined from the MScanFit program. RESULTS: The average MUNE of the anconeus muscle was 55.09 ± 3.27 (mean ± standard error of the mean) for the first test and 54.64 ± 3.70 for the retest, demonstrating excellent measurement reliability, with an intraclass correlation coefficient of 0.90. DISCUSSION: A relatively low motor unit number is found in the anconeus, a muscle not comprehensively studied in literature.

Discovery of N-arylcinnamamides as novel erythroblast enucleation inducers.

Derivation of mature red blood cells (RBCs) from stem cells in vitro is a promising solution to the current shortage of blood supply, in which terminal enucleation is the rate-limiting step. Here we discovered two cinnamamides B8 and B16 showed potential activities of enhancing the enucleation of erythroblasts through the screening of "in-house" compound library. Subsequently, twenty-four N-arylcinnamamides were rationally designed and synthesized on the basis of the structure of B8 and B16, in which N-(9H-carbazol-2-yl)cinnamamide (KS-2) significantly elevated the percentage of reticulocytes in the cultured mouse fetal liver cells in vitro (relative enucleation = 2.43). The underlying mechanism of KS-2 in promoting mouse erythroid enucleation is accelerating the process of cell cycle exit via p53 activation in late stage erythrocytes. These results strongly suggest that compound KS-2 is worthy of further study as a potential erythrocyte enucleation inducer.

Significant Enhancement of the Capacity and Cycling Stability of Lithium-Rich Manganese-Based Layered Cathode Materials via Molybdenum Surface Modification.

Lithium-rich manganese-based layered cathode materials are considered to be one of the best options for next-generation lithium-ion batteries, owing to their ultra-high specific capacity (>250 mAh·g−1) and platform voltage. However, their poor cycling stability, caused by the release of lattice oxygen as well as the electrode/electrolyte side reactions accompanying complex phase transformation, makes it difficult to use this material in practical applications. In this work, we suggest a molybdenum surface modification strategy to improve the electrochemical performance of Li1.2Mn0.54Ni0.13Co0.13O2. The Mo-modified Li1.2Mn0.54Ni0.13Co0.13O2 material exhibits an enhanced discharge specific capacity of up to 290.5 mAh·g−1 (20 mA·g−1) and a capacity retention rate of 82% (300 cycles at 200 mA·g−1), compared with 261.2 mAh·g−1 and a 70% retention rate for the material without Mo modification. The significantly enhanced performance of the modified material can be ascribed to the formation of a Mo-compound-involved nanolayer on the surface of the materials, which effectively lessens the electrolyte corrosion of the cathode, as well as the activation of Mo6+ towards Ni2+/Ni4+ redox couples and the pre-activation of a Mo compound. This study offers a facile and effective strategy to address the poor cyclability of lithium-rich manganese-based layered cathode materials.

[Modified Kaixin San improves memory and synaptic damage of mice with Alzheimer's disease by modulating αCaMKâ ¡-PSD95 protein binding through inhibition of neuroinflammation:a study of mechanism].

To investigated the mechanisms underlying the effects of modified Kaixin San(MKXS) on improving memory and synaptic damage of Alzheimer's disease(AD) mouse model with conditional presenilin 1/2 conditional double knockout(PS cDKO). Specifically, 60 PS cDKO mice(3-3.5 months old) and their age-matched wild-type(WT) littermates were randomized into three groups: WT group(n=20), PS cDKO group(n=20), and PS cDKO+MKXS group(n=20). Mice in WT and PS cDKO groups were fed with standard chow and those in PS cDKO+MKXS group were given chow containing MKXS(at 2.55 g·kg~(-1)) for 60 days. Novel object reco-gnition task was employed to detect the recognition memory of mice, and Western blot to detect the protein levels of synapse-associated proteins in the hippocampus(HPC) of mice, such as NR1, NR2 A, NR2 B, p-αCaMKⅡ, tau, and p-tau. Microglial morphology in the HPC CA1 of mice was observed based on immunohistochemistry. Quantitative real time-PCR(qRT-PCR) was employed to detect the mRNA levels of the pro-inflammatory factors and synapse-associated proteins in the HPC of mice, including COX-2, iNOS, IL-1ß, IL-6, TNF-α, PSD95, NR1, NR2 A, NR2 B, and MAP2. The protein levels of IL-1ß, TNF-α, and IL-6 were tested by enzyme-linked immunosorbent assay(ELISA). The interaction between PSD95 and αCaMKⅡ and between PSD95 and p-αCaMKⅡ was tested by co-immunoprecipitation(Co-IP). The results showed that PS cDKO+MKXS demonstrated significantly higher preference index and recognition index of the new objects, lower protein level of p-tau(ser 396/404) and mRNA levels of COX-2, iNOS, TNF-α, IL-1ß, and IL-6 in HPC, higher protein levels of NR1, NR2 A, NR2 B, and p-αCaMKⅡ and mRNA levels of NR1, NR2 A, NR2 B, PSD95, and MAP2, and stronger interaction of αCaMKⅡ with PSD95 and interaction of p-αCaMKⅡ with PSD95 than the PS cDKO group. Immunohistoche-mical staining showed that MKXS inhibited the activation of microglia. In conclusion, MKXS improves memory and synaptic damage in mice with AD by modulating αCaMKⅡ-PSD95 protein binding through inhibition of neuroinflammation.

Neuronal Glutamatergic Synaptic Clefts Alkalinize Rather Than Acidify during Neurotransmission.

The dogma that the synaptic cleft acidifies during neurotransmission is based on the corelease of neurotransmitters and protons from synaptic vesicles, and is supported by direct data from sensory ribbon-type synapses. However, it is unclear whether acidification occurs at non-ribbon-type synapses. Here we used genetically encoded fluorescent pH indicators to examine cleft pH at conventional neuronal synapses. At the neuromuscular junction of female Drosophila larvae, we observed alkaline spikes of over 1 log unit during fictive locomotion in vivo. Ex vivo, single action potentials evoked alkalinizing pH transients of only ∼0.01 log unit, but these transients summated rapidly during burst firing. A chemical pH indicator targeted to the cleft corroborated these findings. Cleft pH transients were dependent on Ca2+ movement across the postsynaptic membrane, rather than neurotransmitter release per se, a result consistent with cleft alkalinization being driven by the Ca2+/H+ antiporting activity of the plasma membrane Ca2+-ATPase at the postsynaptic membrane. Targeting the pH indicators to the microenvironment of the presynaptic voltage gated Ca2+ channels revealed that alkalinization also occurred within the cleft proper at the active zone and not just within extrasynaptic regions. Application of the pH indicators at the mouse calyx of Held, a mammalian central synapse, similarly revealed cleft alkalinization during burst firing in both males and females. These findings, made at two quite different non-ribbon type synapses, suggest that cleft alkalinization during neurotransmission, rather than acidification, is a generalizable phenomenon across conventional neuronal synapses.SIGNIFICANCE STATEMENT Neurotransmission is highly sensitive to the pH of the extracellular milieu. This is readily evident in the neurological symptoms that accompany systemic acid/base imbalances. Imaging data from sensory ribbon-type synapses show that neurotransmission itself can acidify the synaptic cleft, likely due to the corelease of protons and glutamate. It is not clear whether the same phenomenon occurs at conventional neuronal synapses due to the difficulties in collecting such data. If it does occur, it would provide for an additional layer of activity-dependent modulation of neurotransmission. Our findings of alkalinization, rather than acidification, within the cleft of two different neuronal synapses encourages a reassessment of the scope of activity-dependent pH influences on neurotransmission and short-term synaptic plasticity.