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GNPS Metabolomics Profiling of Single Enlarged Lysosomes Defines the Molecular Signature of Lysosomal Heterogeneity

ABSTRACT: Lysosomes are critical for cellular metabolism and heterogeneously involved in various cellular processes such as endocytosis, autophagy and senescence. The ability to measure lysosomal metabolic heterogeneity is essential for understanding its physiological roles. We therefore built a single-lysosome mass spectrometry (SLMS) platform integrating lysosomal patch-clamp recording and induced nanoESI/MS that enabled concurrent metabolic and electrophysiological profiling of individual enlarged lysosomes.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Primary Mouse Embryonic Fibroblasts (mefs) Human Uroepithelial Cell Line (sv-huc-1 Cells) Hek-293t Mouse Lung Fibroblasts (mlfs) T24 Bladder Carcinoma Cell Line (t24 Cells)

SUBMITTER: Wei Xiong

PROVIDER: MSV000087208 | GNPS | Wed Apr 14 23:18:00 BST 2021

REPOSITORIES: GNPS

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Project description:Arbuscular mycorrhizal (AM) fungi contribute to plant nutrient uptake in systems managed with reduced fertilizer inputs such as organic agriculture and natural ecosystems by extending the effective size of the rhizosphere and delivering mineral. Connecting the molecular study of the AM symbiosis with agriculturally- and ecologically-relevant field environments remains a challenge and is a largely unexplored research topic. This study utilized a cross-disciplinary approach to examine the transcriptional, metabolic, and physiological responses of tomato (Solanum lycopersicum) AM roots to a localized patch of nitrogen (N). A wild-type mycorrhizal tomato and a closely-related nonmycorrhizal mutant were grown at an organic farm in soil that contained an active AM extraradical hyphal network and soil microbe community. The majority of genes regulated by upon enrichment of nitrogen were similarly expressed in mycorrhizal and nonmycorrhizal roots, suggesting that the primary response to an enriched N patch is mediated by mycorrhiza-independent root processes. However where inorganic N concentrations in the soil were low, differential regulation of key tomato N transport and assimilation genes indicate a transcriptome shift towards mycorrhiza-mediated N uptake over direct root supplied N. Furthermore, two novel mycorrhizal-specific tomato ammonium transporters were also found to be regulated under low N conditions. A conceptual model is presented integrating the transcriptome response to low N and highlighting the mycorrhizal-specific ammonium transporters. These results enhance our understanding of the role of the AM symbiosis in sensing and response to an enriched N patch, and demonstrate that transcriptome analyses of complex plant-microbe-soil interactions provide a global snapshot of biological processes relevant to soil processes in organic agriculture. 30 samples were analyzed. There were 2 genotypes (wildtype and mutant) and 3 treatments (two N treatments and a water control) for a total of 6 groups. Each group had 5 biological replicates.

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GNPS Metabolomics Profiling of Single Enlarged Lysosomes Defines the Molecular Signature of Lysosomal Heterogeneity

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