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  • 1
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    NLRP3 inflammasome assembly in neutrophils is supported by PAD4 and promotes NETosis under sterile conditions (2021)
    Frontiers Media
    Details
    Publication Date: 2022-10-27
    Description: © The Author(s), 2021. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Muenzer, P., Negro, R., Fukui, S., di Meglio, L., Aymonnier, K., Chu, L., Cherpokova, D., Gutch, S., Sorvillo, N., Shi, L., Magupalli, V. G., Weber, A. N. R., Scharf, R. E., Waterman, C. M., Wu, H., & Wagner, D. D. NLRP3 inflammasome assembly in neutrophils is supported by PAD4 and promotes NETosis under sterile conditions. Frontiers in Immunology, 12, (2021): 683803, https://doi.org/10.3389/fimmu.2021.683803.
    Description: Neutrophil extracellular trap formation (NETosis) and the NLR family pyrin domain containing 3 (NLRP3) inflammasome assembly are associated with a similar spectrum of human disorders. While NETosis is known to be regulated by peptidylarginine deiminase 4 (PAD4), the role of the NLRP3 inflammasome in NETosis was not addressed. Here, we establish that under sterile conditions the cannonical NLRP3 inflammasome participates in NETosis. We show apoptosis-associated speck-like protein containing a CARD (ASC) speck assembly and caspase-1 cleavage in stimulated mouse neutrophils without LPS priming. PAD4 was needed for optimal NLRP3 inflammasome assembly by regulating NLRP3 and ASC protein levels post-transcriptionally. Genetic ablation of NLRP3 signaling resulted in impaired NET formation, because NLRP3 supported both nuclear envelope and plasma membrane rupture. Pharmacological inhibition of NLRP3 in either mouse or human neutrophils also diminished NETosis. Finally, NLRP3 deficiency resulted in a lower density of NETs in thrombi produced by a stenosis-induced mouse model of deep vein thrombosis. Altogether, our results indicate a PAD4-dependent formation of the NLRP3 inflammasome in neutrophils and implicate NLRP3 in NETosis under noninfectious conditions in vitro and in vivo.
    Description: This work was supported by a grant from National Heart, Lung, and Blood Institute of the National Institutes of Health (grant R35 HL135765) and a Steven Berzin family support to DDW, an Individual Erwin Deutsch fellowship by the German, Austrian and Swiss Society of Thrombosis and Hemostasis Research to RES, a Whitman fellowship (MBL) to DDW, and an Individual Marie Skłodowska-Curie Actions fellowship by the European Commission (796365 - COAGULANT) to PM. ANRW was funded by the Deutsche Forschungsgemeinschaft (TRR156/2 –246807620) and a research grant (We-4195/15-19). CMW was supported by the Division of Intramural Research, NHLBI, NIH.
    Keywords: Neutrophils ; NETs ; NLRP3 inflammasome ; MCC950 ; Deep vein thrombosis ; PAD4
    Repository Name: Woods Hole Open Access Server
    Type: Article
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  • 2
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    Synuclein regulates synaptic vesicle clustering and docking at a vertebrate synapse (2022)
    Frontiers Media
    Details
    Publication Date: 2022-10-27
    Description: © The Author(s), 2021. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Fouke, K. E., Wegman, M. E., Weber, S. A., Brady, E. B., Roman-Vendrell, C., & Morgan, J. R. Synuclein regulates synaptic vesicle clustering and docking at a vertebrate synapse. Frontiers in Cell and Developmental Biology, 9, (2021): 774650, https://doi.org/10.3389/fcell.2021.774650.
    Description: Neurotransmission relies critically on the exocytotic release of neurotransmitters from small synaptic vesicles (SVs) at the active zone. Therefore, it is essential for neurons to maintain an adequate pool of SVs clustered at synapses in order to sustain efficient neurotransmission. It is well established that the phosphoprotein synapsin 1 regulates SV clustering at synapses. Here, we demonstrate that synuclein, another SV-associated protein and synapsin binding partner, also modulates SV clustering at a vertebrate synapse. When acutely introduced to unstimulated lamprey reticulospinal synapses, a pan-synuclein antibody raised against the N-terminal domain of α-synuclein induced a significant loss of SVs at the synapse. Both docked SVs and the distal reserve pool of SVs were depleted, resulting in a loss of total membrane at synapses. In contrast, antibodies against two other abundant SV-associated proteins, synaptic vesicle glycoprotein 2 (SV2) and vesicle-associated membrane protein (VAMP/synaptobrevin), had no effect on the size or distribution of SV clusters. Synuclein perturbation caused a dose-dependent reduction in the number of SVs at synapses. Interestingly, the large SV clusters appeared to disperse into smaller SV clusters, as well as individual SVs. Thus, synuclein regulates clustering of SVs at resting synapses, as well as docking of SVs at the active zone. These findings reveal new roles for synuclein at the synapse and provide critical insights into diseases associated with α-synuclein dysfunction, such as Parkinson’s disease.
    Description: Funding support for this project was provided by the National Institutes of Health NINDS/NIA R01 NS078165 (to JM); University of Chicago Jeff Metcalf Fellowship Grant (to SW).
    Keywords: Exocytosis ; Endocytosis ; Synapsin ; Lamprey ; Liquid phase separation ; VAMP2
    Repository Name: Woods Hole Open Access Server
    Type: Article
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  • 3
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    Small-scale variability dominates benthic coverage and diversity across the Jardines de la Reina, Cuba coral reef system (2020)
    Frontiers Media
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    Publication Date: 2022-10-26
    Description: © The Author(s), 2019. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Hernandez-Fernandez, L., Gonzalez de Zayas, R., Weber, L., Apprill, A., & Armenteros, M. Small-scale variability dominates benthic coverage and diversity across the Jardines de la Reina, Cuba coral reef system. Frontiers in Marine Science, 6, (2019): 747, doi: 10.3389/fmars.2019.00747.
    Description: Coral reefs are complex and biodiverse ecosystems that are undergoing significant change. Understanding reef composition and biodiversity at multiple spatial scales is necessary to track both large-scale and more subtle ecosystem changes. The Jardines de la Reina (JR) archipelago, located offshore of the island of Cuba, contains the largest marine protected area (MPA) in the Caribbean Sea but lacks multi-scale studies. In this contribution, we documented the diversity of scleractinian corals, octocorals, algae, and sponges across nested spatial scales spanning four orders of magnitude (101–105 m). In addition, we tested the hypothesis that species diversity followed a gradient along the ca. 200 km of reef tract. Across the archipelago, we examined benthic cover and species diversity within 255 photo-quadrats (25 × 25 cm) at 13 fore reef sites (two sampling locations per site, and 10 photo-quadrats per location). Small-scale (101 m) variability between photo-quadrats characterized the coral reef community structure in JR compared with local- (102 m) and mesoscale (104–105 m) variability. This finding suggests that biological processes (e.g., recruitment, competition) had primacy over hydrodynamics for driving the differences in reef community composition. However, the dominance of algae and low cover and diversity of scleractinian corals suggests the pervasive effects of global change on coral communities despite potential benefits provided by the MPA (e.g., oligotrophy and abundance of herbivores). There was no gradient of benthic community structure along the fore reef tract of JR; instead, a patchy distribution occurred in response to more subtle drivers acting at local scales. Overall, our multi-scale comparison was useful for differentiating the impacts of processes potentially impacting the JR reefs, thus providing important information to understand how reef communities are impacted by different environmental and anthropogenic stressors, and the potential benefits of MPAs.
    Description: This work was supported by the Dalio Foundation’s Dalio Ocean Initiative (now “OceanX”).
    Keywords: Coral reef ; Caribbean Sea ; Protected area ; Species richness ; β-diversity ; Spatial scale
    Repository Name: Woods Hole Open Access Server
    Type: Article
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  • 4
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    New Calcium Carbonate Nano‐particulate Pressed Powder Pellet (NFHS‐2‐NP) for LA‐ICP‐OES, LA‐(MC)‐ICP‐MS and µXRF (2022)
    geostandard and geoanalytical research
    In:  EPIC3Geostandards and Geoanalytical Research, geostandard and geoanalytical research, ISSN: 1639-4488
    Details
    Publication Date: 2022-06-02
    Repository Name: EPIC Alfred Wegener Institut
    Type: Article , peerRev
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  • 5
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    Extracellular reef metabolites across the protected Jardines de la Reina, Cuba Reef System (2021)
    Frontiers Media
    Details
    Publication Date: 2022-10-26
    Description: © The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Weber, L., Armenteros, M., Soule, M. K., Longnecker, K., Kujawinski, E. B., & Apprill, A. Extracellular reef metabolites across the protected Jardines de la Reina, Cuba Reef System. Frontiers in Marine Science, 7, (2020): 582161, https://doi.org/10.3389/fmars.2020.582161.
    Description: Coral reef ecosystems are incredibly diverse marine biomes that rely on nutrient cycling by microorganisms to sustain high productivity in generally oligotrophic regions of the ocean. Understanding the composition of extracellular reef metabolites in seawater, the small organic molecules that serve as the currency for microorganisms, may provide insight into benthic-pelagic coupling as well as the complexity of nutrient cycling in coral reef ecosystems. Jardines de la Reina (JR), Cuba is an ideal environment to examine extracellular metabolites across protected and high-quality reefs. Here, we used liquid chromatography mass spectrometry (LC-MS) to quantify specific known metabolites of interest (targeted metabolomics approach) and to survey trends in metabolite feature composition (untargeted metabolomics approach) from surface and reef depth (6 – 14 m) seawater overlying nine forereef sites in JR. We found that untargeted metabolite feature composition was surprisingly similar between reef depth and surface seawater, corresponding with other biogeochemical and physicochemical measurements and suggesting that environmental conditions were largely homogenous across forereefs within JR. Additionally, we quantified 32 of 53 detected metabolites using the targeted approach, including amino acids, nucleosides, vitamins, and other metabolic intermediates. Two of the quantified metabolites, riboflavin and xanthosine, displayed interesting trends by depth. Riboflavin concentrations were higher in reef depth compared to surface seawater, suggesting that riboflavin may be produced by reef organisms at depth and degraded in the surface through photochemical oxidation. Xanthosine concentrations were significantly higher in surface reef seawater. 5′-methylthioadenosine (MTA) concentrations increased significantly within the central region of the archipelago, displaying biogeographic patterns that warrant further investigation. Here we lay the groundwork for future investigations of variations in metabolite composition across reefs, sources and sinks of reef metabolites, and changes in metabolites over environmental, temporal, and reef health gradients.
    Description: This work was supported by the Dalio Foundation (now “OceanX”) and the National Science Foundation (OCE-1736288) (award to Amy Apprill). The mass spectrometry samples were analyzed at the WHOI FT-MS Users’ Facility with instrumentation funded by the National Science Foundation (grant OCE-1058448 to EK and MK) and the Simons Foundation (Award ID #509042, EK). Lastly, a portion of the publication fees was supported by the Massachusetts Institute of Technology (MIT) Open Access Article Publication Subvention fund from MIT Libraries.
    Keywords: Metabolomics ; Coral reefs ; Microorganisms ; Ecology ; DOM cycling
    Repository Name: Woods Hole Open Access Server
    Type: Article
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