title: Partial rescue of glomerular laminin alpha5 mutations by wild-type endothelia produce hybrid glomeruli.
abstract: Both endothelial cells and podocytes are sources for laminin alpha1 at the inception of glomerulogenesis and then for laminin alpha5 during glomerular maturation. Why glomerular basement membranes (GBM) undergo laminin transitions is unknown, but this may dictate glomerular morphogenesis. In mice that genetically lack laminin alpha5, laminin alpha5beta2gamma1 is not assembled, vascularized glomeruli fail to form, and animals die at midgestation with neural tube closure and placental deficits. It was previously shown that renal cortices of newborn mice contain endothelial progenitors (angioblasts) and that when embryonic day 12 kidneys are transplanted into newborn kidney, hybrid glomeruli (host-derived endothelium and donor-derived podocytes) result. Reasoning that host endothelium may correct the glomerular phenotype that is seen in laminin alpha5 mutants, alpha5 null embryonic day 12 metanephroi were grafted into wild-type newborn kidney. Hybrid glomeruli were identified in grafts by expression of a host-specific LacZ lineage marker. Labeling of glomerular hybrid GBM with chain-specific antibodies showed a markedly stratified distribution of laminins: alpha5 was found only on the inner endothelial half of GBM, whereas alpha1 located to outer layers beneath mutant podocytes. For measurement of the contribution of host endothelium to hybrid GBM, immunofluorescent signals for laminin alpha5 were quantified: Hybrid GBM contained approximately 50% the normal alpha5 complement as wild-type GBM. Electron microscopy of glomerular hybrids showed vascularization, but podocyte foot processes were absent. It was concluded that (1) endothelial and podocyte-derived laminins remain tethered to their cellular origin, (2) developing endothelial cells contribute large amounts of GBM laminins, and (3) podocyte foot process differentiation may require direct exposure to laminin alpha5.
authors.affiliation: Department of Anatomy and Cell Biology, University of Kansas Medical Center, MS 3038, 3901 Rainbow Boulevard, Kansas City, KS 66160, USA. dabrahamson@kumc.edu
authors.forename: Dale R
authors.initials: DR
authors.lastname: Abrahamson
authors.forename: Patricia L
authors.initials: PL
authors.lastname: St John
authors.forename: Kathryn
authors.initials: K
authors.lastname: Isom
authors.forename: Barry
authors.initials: B
authors.lastname: Robert
authors.forename: Jeffrey H
authors.initials: JH
authors.lastname: Miner
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002454:Cell Differentiation / Q000502:physiology; D002678:Chimera; D042783:Endothelial Cells / Q000502:physiology / Q000648:ultrastructure; D016332:Fetal Tissue Transplantation; D018507:Gene Expression Regulation, Developmental; D007678:Kidney Glomerulus / Q000166:cytology / Q000254:growth & development* / Q000637:transplantation*; D007797:Laminin / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D008815:Mice, Inbred Strains; D018345:Mice, Knockout; D008854:Microscopy, Electron; D010641:Phenotype; D050199:Podocytes / Q000502:physiology* / Q000648:ultrastructure; D014184:Transplantation, Homologous
chemical_list: D007797:Laminin; C097043:laminin alpha5