title: Glomerular cell cross-talk influences composition and assembly of extracellular matrix.
abstract: The glomerular basement membrane (GBM) is a specialized extracellular matrix (ECM) compartment within the glomerulus that contains tissue-restricted isoforms of collagen IV and laminin. It is integral to the capillary wall and therefore, functionally linked to glomerular filtration. Although the composition of the GBM has been investigated with global and candidate-based approaches, the relative contributions of glomerular cell types to the production of ECM are not well understood. To characterize specific cellular contributions to the GBM, we used mass spectrometry-based proteomics to analyze ECM isolated from podocytes and glomerular endothelial cells in vitro. These analyses identified cell type-specific differences in ECM composition, indicating distinct contributions to glomerular ECM assembly. Coculture of podocytes and endothelial cells resulted in an altered composition and organization of ECM compared with monoculture ECMs, and electron microscopy revealed basement membrane-like ECM deposition between cocultured cells, suggesting the involvement of cell-cell cross-talk in the production of glomerular ECM. Notably, compared with monoculture ECM proteomes, the coculture ECM proteome better resembled a tissue-derived glomerular ECM dataset, indicating its relevance to GBM in vivo. Protein network analyses revealed a common core of 35 highly connected structural ECM proteins that may be important for glomerular ECM assembly. Overall, these findings show the complexity of the glomerular ECM and suggest that both ECM composition and organization are context-dependent.
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences and.
authors.forename: Adam
authors.initials: A
authors.lastname: Byron
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences and Faculty of Medical and Human Sciences, University of Manchester, Manchester, United Kingdom;
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Randles
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences and.
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Humphries
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences and.
authors.forename: Aleksandr
authors.initials: A
authors.lastname: Mironov
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences and.
authors.forename: Hellyeh
authors.initials: H
authors.lastname: Hamidi
authors.affiliation: Faculty of Medical and Human Sciences, University of Manchester, Manchester, United Kingdom;
authors.forename: Shelley
authors.initials: S
authors.lastname: Harris
authors.affiliation: Academic Renal Unit, Faculty of Medicine and Dentistry, University of Bristol, Bristol, United Kingdom;
authors.forename: Peter W
authors.initials: PW
authors.lastname: Mathieson
authors.affiliation: Academic Renal Unit, Faculty of Medicine and Dentistry, University of Bristol, Bristol, United Kingdom;
authors.forename: Moin A
authors.initials: MA
authors.lastname: Saleem
authors.affiliation: Academic Renal Unit, Faculty of Medicine and Dentistry, University of Bristol, Bristol, United Kingdom;
authors.forename: Simon C
authors.initials: SC
authors.lastname: Satchell
authors.affiliation: Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee; and Department of Medicine, Veterans Affairs Hospital, Nashville, Tennessee.
authors.forename: Roy
authors.initials: R
authors.lastname: Zent
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences and.
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Humphries
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences and Faculty of Medical and Human Sciences, University of Manchester, Manchester, United Kingdom; Rachel.Lennon@manchester.ac.uk.
authors.forename: Rachel
authors.initials: R
authors.lastname: Lennon
mesh_terms: D002478:Cells, Cultured; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D005109:Extracellular Matrix / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis / Q000502:physiology*; D006801:Humans; D007678:Kidney Glomerulus / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D010641:Phenotype; D050199:Podocytes / Q000502:physiology; D060066:Protein Interaction Maps; D020239:Receptor Cross-Talk / Q000502:physiology*
chemical_list: D017077:Culture Media, Conditioned; D016326:Extracellular Matrix Proteins