title: Glomerular expression of transforming growth factor-beta (TGF-beta) isoforms in mice lacking CD2-associated protein.
abstract: Mice lacking CD2-associated protein (CD2AP-/-) develop glomerular lesions resembling human focal segmental glomerulosclerosis (FSGS) between 3-4 weeks of age and die approximately 2 weeks later from massive proteinuria and renal failure. The mechanisms involved in the glomerular injury in this model are unclear. In this study, we used laser capture microdissection (LCM) and real-time PCR, and examined expression of TGF-ss isoforms in CD2AP-/- mice at the level of isolated glomeruli. Total RNA yield from cryosections of 30 glomeruli was 10.71 ng (SD, 5.45) in CD2AP+/+ group (n =7), and 4.20 ng (SD, 2.04) in CD2AP-/- group (n =8), p =0.008. Expression of TGF-ss1 mRNA was increased 1.5-fold in the whole kidney (p =0.030), and twofold in isolated CD2AP-/- glomeruli (p =0.026). Whole kidney mRNA of TGF-ss receptor I (RI) and II (RII) was not different in CD2AP-/- and CD2AP+/+ animals, but it was increased in CD2AP-/- glomerular samples by 4.38-fold (p =0.001) and 11.37-fold (p =0.0163), respectively. By using LCM we confirmed increased glomerular expression levels of TGF-ss isoforms previously described by our group in glomeruli isolated by sieving in CD2AP KO mice and underscored the importance of local factors in the development of glomerulosclerosis.
authors.affiliation: Section of Pediatric Nephrology, Children's Hospital at Montefiore, Albert Einstein College of Medicine, Bronx, NY 10467, USA.
authors.forename: Robert P
authors.initials: RP
authors.lastname: Woroniecki
authors.forename: Mario
authors.initials: M
authors.lastname: Schiffer
authors.forename: Andrey S
authors.initials: AS
authors.lastname: Shaw
authors.forename: Frederick J
authors.initials: FJ
authors.lastname: Kaskel
authors.forename: Erwin P
authors.initials: EP
authors.lastname: Bottinger
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D003598:Cytoskeletal Proteins; D015870:Gene Expression; D005923:Glomerulosclerosis, Focal Segmental / Q000378:metabolism* / Q000473:pathology; D007678:Kidney Glomerulus / Q000378:metabolism* / Q000473:pathology; D007834:Lasers; D051379:Mice; D018345:Mice, Knockout; D042282:Microdissection; D050199:Podocytes / Q000473:pathology; D020033:Protein Isoforms; D011506:Proteins / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis; D018125:Receptors, Transforming Growth Factor beta / Q000378:metabolism; D015398:Signal Transduction; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D025721:WT1 Proteins / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C114545:CD2-associated protein; D003598:Cytoskeletal Proteins; D020033:Protein Isoforms; D011506:Proteins; D012333:RNA, Messenger; D018125:Receptors, Transforming Growth Factor beta; D016212:Transforming Growth Factor beta; D025721:WT1 Proteins