title: Overexpression of TGF-β Inducible microRNA-143 in Zebrafish Leads to Impairment of the Glomerular Filtration Barrier by Targeting Proteoglycans.
abstract: BACKGROUND
  TGF-β is known as an important stress factor of podocytes in glomerular diseases. Apart from activation of direct pro-apoptotic pathways we wanted to analyze micro-RNA (miRs) driven regulation of components involved in the integrity of the glomerular filtration barrier induced by TGF-β. Since miR-143-3p (miR-143) is described as a TGF-β inducible miR in other cell types, we examined this specific miR and its ability to induce glomerular pathology.
  
  
  METHODS
  We analyzed miR-143 expression in cultured human podocytes after stimulation with TGF-β. We also microinjected zebrafish eggs with a miR-143 mimic or with morpholinos specific for its targets syndecan and versican and compared phenotype and proteinuria development.
  
  
  RESULTS
  We detected a time dependent, TGF-β inducible expression of miR-143 in human podocytes. Targets of miR-143 relevant in glomerular biology are syndecans and versican, which are known components of the glycocalyx. We found that syndecan 1 and 4 were predominantly expressed in podocytes while syndecan 3 was largely expressed in glomerular endothelial cells. Versican could be detected in both cell types. After injection of a miR-143 mimic in zebrafish larvae, syndecan 3, 4 and versican were significantly downregulated. Moreover, miR-143 overexpression or versican knockdown by morpholino caused loss of plasma proteins, edema, podocyte effacement and endothelial damage. In contrast, knockdown of syndecan 3 and syndecan 4 had no effects on glomerular filtration barrier.
  
  
  CONCLUSIONS
  Expression of versican and syndecan isoforms is indispensable for proper barrier function. Podocyte-derived miR-143 is a mediator for paracrine and autocrine cross talk between podocytes and glomerular endothelial cells and can alter expression of glomerular glycocalyx proteins.
authors.affiliation: Department of Medicine/Nephrology, Hannover Medical School, Hannover ,Germany.
authors.forename: Janina
authors.initials: J
authors.lastname: Müller-Deile
authors.forename: Finn
authors.initials: F
authors.lastname: Gellrich
authors.forename: Heiko
authors.initials: H
authors.lastname: Schenk
authors.forename: Patricia
authors.initials: P
authors.lastname: Schroder
authors.forename: Jenny
authors.initials: J
authors.lastname: Nyström
authors.forename: Johan
authors.initials: J
authors.lastname: Lorenzen
authors.forename: Hermann
authors.initials: H
authors.lastname: Haller
authors.forename: Mario
authors.initials: M
authors.lastname: Schiffer
mesh_terms: D000818:Animals; D001483:Base Sequence; D001798:Blood Proteins / Q000378:metabolism; D002478:Cells, Cultured; D004487:Edema / Q000473:pathology; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D055785:Gene Knockdown Techniques; D060730:Glomerular Filtration Barrier / Q000187:drug effects / Q000473:pathology*; D019276:Glycocalyx / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007678:Kidney Glomerulus / Q000187:drug effects / Q000473:pathology; D007814:Larva / Q000187:drug effects / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D060172:Morpholinos / Q000494:pharmacology; D050199:Podocytes / Q000187:drug effects / Q000378:metabolism; D011507:Proteinuria / Q000378:metabolism / Q000473:pathology; D011509:Proteoglycans / Q000378:metabolism*; D053667:Syndecans / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics; D053675:Versicans / Q000378:metabolism; D015027:Zebrafish / Q000235:genetics*; D029961:Zebrafish Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D001798:Blood Proteins; C549612:MIRN143 microRNA, zebrafish; D035683:MicroRNAs; D060172:Morpholinos; D011509:Proteoglycans; D053667:Syndecans; D016212:Transforming Growth Factor beta; D029961:Zebrafish Proteins; D053675:Versicans