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      Adipose triglyceride lipase protects renal cell endocytosis in a Drosophila dietary model of chronic kidney disease

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          Abstract

          Obesity-related renal lipotoxicity and chronic kidney disease (CKD) are prevalent pathologies with complex aetiologies. One hallmark of renal lipotoxicity is the ectopic accumulation of lipid droplets in kidney podocytes and in proximal tubule cells. Renal lipid droplets are observed in human CKD patients and in high-fat diet (HFD) rodent models, but their precise role remains unclear. Here, we establish a HFD model in Drosophila that recapitulates renal lipid droplets and several other aspects of mammalian CKD. Cell type–specific genetic manipulations show that lipid can overflow from adipose tissue and is taken up by renal cells called nephrocytes. A HFD drives nephrocyte lipid uptake via the multiligand receptor Cubilin (Cubn), leading to the ectopic accumulation of lipid droplets. These nephrocyte lipid droplets correlate with endoplasmic reticulum (ER) and mitochondrial deficits, as well as with impaired macromolecular endocytosis, a key conserved function of renal cells. Nephrocyte knockdown of diglyceride acyltransferase 1 (DGAT1), overexpression of adipose triglyceride lipase (ATGL), and epistasis tests together reveal that fatty acid flux through the lipid droplet triglyceride compartment protects the ER, mitochondria, and endocytosis of renal cells. Strikingly, boosting nephrocyte expression of the lipid droplet resident enzyme ATGL is sufficient to rescue HFD-induced defects in renal endocytosis. Moreover, endocytic rescue requires a conserved mitochondrial regulator, peroxisome proliferator-activated receptor-gamma coactivator 1α (PGC1α). This study demonstrates that lipid droplet lipolysis counteracts the harmful effects of a HFD via a mitochondrial pathway that protects renal endocytosis. It also provides a genetic strategy for determining whether lipid droplets in different biological contexts function primarily to release beneficial or to sequester toxic lipids.

          Abstract

          A high-fat diet model of chronic kidney disease in Drosophila reveals that boosting triglyceride lipolysis in renal cells is sufficient to rescue renal cell function via a pathway involving PGC1 alpha and mitochondria.

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          Fiji is a distribution of the popular open-source software ImageJ focused on biological-image analysis. Fiji uses modern software engineering practices to combine powerful software libraries with a broad range of scripting languages to enable rapid prototyping of image-processing algorithms. Fiji facilitates the transformation of new algorithms into ImageJ plugins that can be shared with end users through an integrated update system. We propose Fiji as a platform for productive collaboration between computer science and biology research communities.
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              A guided tour into subcellular colocalization analysis in light microscopy.

              It is generally accepted that the functional compartmentalization of eukaryotic cells is reflected by the differential occurrence of proteins in their compartments. The location and physiological function of a protein are closely related; local information of a protein is thus crucial to understanding its role in biological processes. The visualization of proteins residing on intracellular structures by fluorescence microscopy has become a routine approach in cell biology and is increasingly used to assess their colocalization with well-characterized markers. However, image-analysis methods for colocalization studies are a field of contention and enigma. We have therefore undertaken to review the most currently used colocalization analysis methods, introducing the basic optical concepts important for image acquisition and subsequent analysis. We provide a summary of practical tips for image acquisition and treatment that should precede proper colocalization analysis. Furthermore, we discuss the application and feasibility of colocalization tools for various biological colocalization situations and discuss their respective strengths and weaknesses. We have created a novel toolbox for subcellular colocalization analysis under ImageJ, named JACoP, that integrates current global statistic methods and a novel object-based approach.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Formal analysisRole: InvestigationRole: MethodologyRole: ValidationRole: Writing – original draftRole: Writing – review & editing
                Role: ConceptualizationRole: InvestigationRole: MethodologyRole: SupervisionRole: Writing – review & editing
                Role: Formal analysisRole: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: Formal analysisRole: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: Formal analysisRole: ValidationRole: Writing – review & editing
                Role: Formal analysisRole: InvestigationRole: MethodologyRole: ValidationRole: Writing – review & editing
                Role: Formal analysis
                Role: MethodologyRole: ResourcesRole: SupervisionRole: Writing – review & editing
                Role: ConceptualizationRole: Formal analysisRole: Funding acquisitionRole: MethodologyRole: ResourcesRole: SupervisionRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: Academic Editor
                Journal
                PLoS Biol
                PLoS Biol
                plos
                plosbiol
                PLoS Biology
                Public Library of Science (San Francisco, CA USA )
                1544-9173
                1545-7885
                4 May 2021
                May 2021
                4 May 2021
                : 19
                : 5
                : e3001230
                Affiliations
                [1 ] Physiology and Metabolism Laboratory, The Francis Crick Institute, London, United Kingdom
                [2 ] Electron Microscopy Science Technology Platform, The Francis Crick Institute, London, United Kingdom
                University of Cambridge, UNITED KINGDOM
                Author notes

                The authors have declared that no competing interests exist.

                [¤]

                Current address: Wellcome Centre for Human Genetics, University of Oxford, Oxford, United Kingdom

                Author information
                https://orcid.org/0000-0002-4796-5884
                https://orcid.org/0000-0001-8015-1420
                https://orcid.org/0000-0003-2143-1689
                https://orcid.org/0000-0003-2667-7426
                https://orcid.org/0000-0002-6067-5160
                https://orcid.org/0000-0003-0260-613X
                https://orcid.org/0000-0002-6875-6985
                Article
                PBIOLOGY-D-20-02824
                10.1371/journal.pbio.3001230
                8121332
                33945525
                521530f5-5b15-4158-8e11-3d13dd0e3f8f
                © 2021 Lubojemska et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 21 September 2020
                : 13 April 2021
                Page count
                Figures: 6, Tables: 0, Pages: 25
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/100004440, Wellcome Trust;
                Award ID: 104566/Z/14/Z
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100010438, Francis Crick Institute;
                Award ID: FC001088
                Award Recipient :
                This work was supported by an Investigator Award to APG from the Wellcome Trust (104566/Z/14/Z, https://wellcome.org) and by funding to APG from the Francis Crick Institute ( https://www.crick.ac.uk), which receives its core funding from Cancer Research UK (FC001088), the UK Medical Research Council (FC001088) and the Wellcome Trust (FC001088). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
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                Biochemistry
                Lipids
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                2021-05-14
                All relevant data are within the paper and its Supporting Information files. Statistical analysis values are provided in S1 Data and the source data for all main and supporting graphs are provided in S2 Data.

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