Skip to main content
Top
Published in: Pediatric Nephrology 3/2006

01-03-2006 | Original Article

Glomerular expression of transforming growth factor–β (TGF-β) isoforms in mice lacking CD2-associated protein

Authors: Robert P. Woroniecki, Mario Schiffer, Andrey S. Shaw, Frederick J. Kaskel, Erwin P. Bottinger

Published in: Pediatric Nephrology | Issue 3/2006

Login to get access

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.
Literature
1.
go back to reference Warady BA, Hebert D, Sullivan EK, Alexander SR, Tejani A (1997) Renal transplantation, chronic dialysis, and chronic renal insufficiency in children and adolescents. The 1995 Annual Report of the North American Pediatric Renal Transplant Cooperative Study. Pediatr Nephrol 11:49–64CrossRefPubMed Warady BA, Hebert D, Sullivan EK, Alexander SR, Tejani A (1997) Renal transplantation, chronic dialysis, and chronic renal insufficiency in children and adolescents. The 1995 Annual Report of the North American Pediatric Renal Transplant Cooperative Study. Pediatr Nephrol 11:49–64CrossRefPubMed
2.
go back to reference Braden GL (2000) Changing incidence of glomerular diseases in adults. Am J Kidney Dis 35(5):878–883PubMed Braden GL (2000) Changing incidence of glomerular diseases in adults. Am J Kidney Dis 35(5):878–883PubMed
3.
go back to reference Kriz W, Lemley KV (1999) The role of the podocyte in glomerulosclerosis. Curr Opin Nephrol Hypertens 8(4):489–497CrossRefPubMed Kriz W, Lemley KV (1999) The role of the podocyte in glomerulosclerosis. Curr Opin Nephrol Hypertens 8(4):489–497CrossRefPubMed
4.
go back to reference Hattori M, Horita S, Yoshioka T, Yamaguchi Y, Kawaguchi H, Ito K(1997) Mesangial phenotypic changes associated with cellular lesions in primary focal segmental glomerulosclerosis. Am J Kidney Dis 30:632–638PubMed Hattori M, Horita S, Yoshioka T, Yamaguchi Y, Kawaguchi H, Ito K(1997) Mesangial phenotypic changes associated with cellular lesions in primary focal segmental glomerulosclerosis. Am J Kidney Dis 30:632–638PubMed
5.
go back to reference Mundel P, Shankland SJ (2002) Podocyte biology and response to injury. J Am Soc Nephrol 13:3005–3015CrossRefPubMed Mundel P, Shankland SJ (2002) Podocyte biology and response to injury. J Am Soc Nephrol 13:3005–3015CrossRefPubMed
6.
go back to reference Kim YH, Goyal M, Kurnit D, Wharram B, Wiggins J, Holzman L, Kershaw D, Wiggins R (2001) Podocyte depletion and glomerulosclerosis have a direct relationship in the PAN-treated rat. Kidney Int 60:957–968CrossRefPubMed Kim YH, Goyal M, Kurnit D, Wharram B, Wiggins J, Holzman L, Kershaw D, Wiggins R (2001) Podocyte depletion and glomerulosclerosis have a direct relationship in the PAN-treated rat. Kidney Int 60:957–968CrossRefPubMed
7.
go back to reference Dustin ML, Olszowy MW, Holdorf AD, Li J, Bromley S, Desai N, Widder P, Rosenberger F, van der Merwe PA, Allen PM, Shaw AS (1998) A novel adaptor protein orchestrates receptor patterning and cytoskeletal polarity in T-cell contacts. Cell 94:667–677CrossRefPubMed Dustin ML, Olszowy MW, Holdorf AD, Li J, Bromley S, Desai N, Widder P, Rosenberger F, van der Merwe PA, Allen PM, Shaw AS (1998) A novel adaptor protein orchestrates receptor patterning and cytoskeletal polarity in T-cell contacts. Cell 94:667–677CrossRefPubMed
8.
go back to reference Shih NY, Li J, Karpitskii V, Nguyen A, Dustin ML, Kanagawa O, Miner JH, Shaw AS (1999) Congenital nephrotic syndrome in mice lacking CD2-associated protein. Science 286:312–315CrossRefPubMed Shih NY, Li J, Karpitskii V, Nguyen A, Dustin ML, Kanagawa O, Miner JH, Shaw AS (1999) Congenital nephrotic syndrome in mice lacking CD2-associated protein. Science 286:312–315CrossRefPubMed
9.
go back to reference Kim JM, Wu H, Green G, Winkler CA, Kopp JB, Miner JH, Unanue ER, Shaw AS (2003) CD2-associated protein haploinsufficiency is linked to glomerular disease susceptibility. Science 300:1298–1300CrossRefPubMed Kim JM, Wu H, Green G, Winkler CA, Kopp JB, Miner JH, Unanue ER, Shaw AS (2003) CD2-associated protein haploinsufficiency is linked to glomerular disease susceptibility. Science 300:1298–1300CrossRefPubMed
10.
go back to reference Schwarz K, Simons M, Reiser J, Saleem MA, Faul C, Kriz W, Shaw AS, Holzman LB, Mundel P (2001) Podocin, a raft-associated component of the glomerular slit diaphragm, interacts with CD2AP and nephrin. J Clin Invest 108:1621–1629CrossRefPubMed Schwarz K, Simons M, Reiser J, Saleem MA, Faul C, Kriz W, Shaw AS, Holzman LB, Mundel P (2001) Podocin, a raft-associated component of the glomerular slit diaphragm, interacts with CD2AP and nephrin. J Clin Invest 108:1621–1629CrossRefPubMed
11.
go back to reference Huber TB, Hartleben B, Kim J, Schmidts M, Schermer B, Keil A, Egger L, Lecha RL, Borner C, Pavenstadt H, Shaw AS, Walz G, Benzing T (2003) Nephrin and CD2AP associate with phosphoinositide 3-OH kinase and stimulate AKT-dependent signaling. Mol Cell Biol 23:4917–4928CrossRefPubMed Huber TB, Hartleben B, Kim J, Schmidts M, Schermer B, Keil A, Egger L, Lecha RL, Borner C, Pavenstadt H, Shaw AS, Walz G, Benzing T (2003) Nephrin and CD2AP associate with phosphoinositide 3-OH kinase and stimulate AKT-dependent signaling. Mol Cell Biol 23:4917–4928CrossRefPubMed
12.
13.
go back to reference Border WA, Okuda S, Languino LR, Sporn MB, Ruoslahti E (1990) Suppression of experimental glomerulonephritis by antiserum against transforming growth factor beta 1. Nature 346:371–374CrossRefPubMed Border WA, Okuda S, Languino LR, Sporn MB, Ruoslahti E (1990) Suppression of experimental glomerulonephritis by antiserum against transforming growth factor beta 1. Nature 346:371–374CrossRefPubMed
14.
go back to reference Yamamoto T, Nakamura T, Noble NA, Ruoslahti E, Border WA (1993) Expression of transforming growth factor β is elevated in human and experimental diabetic nephropathy. Proc Natl Acad Sci USA 90:1814–1818PubMed Yamamoto T, Nakamura T, Noble NA, Ruoslahti E, Border WA (1993) Expression of transforming growth factor β is elevated in human and experimental diabetic nephropathy. Proc Natl Acad Sci USA 90:1814–1818PubMed
15.
go back to reference Kopp JB, Factor VM, Mozes M, Nagy P, Sanderson N, Bottinger EP, Klotman PE, Thorgeirsson SS (1996) Transgenic mice with increased plasma levels of TGF-beta 1 develop progressive renal disease. Lab Invest 74:991–1003PubMed Kopp JB, Factor VM, Mozes M, Nagy P, Sanderson N, Bottinger EP, Klotman PE, Thorgeirsson SS (1996) Transgenic mice with increased plasma levels of TGF-beta 1 develop progressive renal disease. Lab Invest 74:991–1003PubMed
16.
go back to reference Schiffer M, Bitzer M, Roberts IS, Kopp JB, ten Dijke P, Mundel P, Bottinger EP (2001) Apoptosis in podocytes induced by TGF-beta and Smad7. J Clin Invest 108:807–816CrossRefPubMed Schiffer M, Bitzer M, Roberts IS, Kopp JB, ten Dijke P, Mundel P, Bottinger EP (2001) Apoptosis in podocytes induced by TGF-beta and Smad7. J Clin Invest 108:807–816CrossRefPubMed
17.
18.
go back to reference Schiffer M, Mundel P, Shaw AS, Bottinger EP (2004) A novel role for the adaptor molecule CD2-associated protein in TGF-beta-induced apoptosis. J Biol Chem 279:37004–37012CrossRefPubMed Schiffer M, Mundel P, Shaw AS, Bottinger EP (2004) A novel role for the adaptor molecule CD2-associated protein in TGF-beta-induced apoptosis. J Biol Chem 279:37004–37012CrossRefPubMed
19.
go back to reference Bonner RF, Emmert-Buck M, Cole K, Pohida T, Chuaqui R, Goldstein S, Liotta LA (1997) Laser capture microdissection: Molecular analysis of tissue. Science 278:1481–1483CrossRefPubMed Bonner RF, Emmert-Buck M, Cole K, Pohida T, Chuaqui R, Goldstein S, Liotta LA (1997) Laser capture microdissection: Molecular analysis of tissue. Science 278:1481–1483CrossRefPubMed
20.
go back to reference Kohda Y, Murakami H, Moe OW, Star RA (2000) Analysis of segmental renal gene expression by laser capture microdissection. Kidney Int 57:321–331CrossRefPubMed Kohda Y, Murakami H, Moe OW, Star RA (2000) Analysis of segmental renal gene expression by laser capture microdissection. Kidney Int 57:321–331CrossRefPubMed
21.
go back to reference Peterson KS, Huang JF, Zhu J, D’Agati V, Liu X, Miller N, Erlander MG, Jackson MR, Winchester RJ (2004) Characterization of heterogeneity in the molecular pathogenesis of lupus nephritis from transcriptional profiles of laser-captured glomeruli. J Clin Invest 113:1722–1733CrossRefPubMed Peterson KS, Huang JF, Zhu J, D’Agati V, Liu X, Miller N, Erlander MG, Jackson MR, Winchester RJ (2004) Characterization of heterogeneity in the molecular pathogenesis of lupus nephritis from transcriptional profiles of laser-captured glomeruli. J Clin Invest 113:1722–1733CrossRefPubMed
22.
go back to reference Schiffer M, Wu DT, Soto-Nieves N, Yu L, Shaw AS, Bottinger EP (2002) Phenotype characterization and gene expression profiling in CD2AP-knockout mice. J Am Soc Nephrol 13:43ACrossRef Schiffer M, Wu DT, Soto-Nieves N, Yu L, Shaw AS, Bottinger EP (2002) Phenotype characterization and gene expression profiling in CD2AP-knockout mice. J Am Soc Nephrol 13:43ACrossRef
23.
go back to reference Craven RA, Totty N, Harnden P, Selby PJ, Banks RE (2002) Laser capture microdissection and two-dimensional polyacrylamide gel electrophoresis. Evaluation of tissue preparation and sample limitations. Am J Pathol 160:815–822PubMed Craven RA, Totty N, Harnden P, Selby PJ, Banks RE (2002) Laser capture microdissection and two-dimensional polyacrylamide gel electrophoresis. Evaluation of tissue preparation and sample limitations. Am J Pathol 160:815–822PubMed
24.
go back to reference Zavadil J, Cermak L, Soto-Nieves N, Bottinger EP (2004) Integration of TGF-beta/Smad and jagged1/notch signalling in epithelial-to-mesenchymal transition. EMBO J 23:1155–1165CrossRefPubMed Zavadil J, Cermak L, Soto-Nieves N, Bottinger EP (2004) Integration of TGF-beta/Smad and jagged1/notch signalling in epithelial-to-mesenchymal transition. EMBO J 23:1155–1165CrossRefPubMed
25.
go back to reference Morrissey J, Guo G, Moridaira K, Fitzgerald M, McCracken R, Tolley T, Klahr S (2002) Transforming growth factor-beta induces renal epithelial jagged-1 expression in fibrotic disease. J Am Soc Nephrol 13:1499–1508CrossRefPubMed Morrissey J, Guo G, Moridaira K, Fitzgerald M, McCracken R, Tolley T, Klahr S (2002) Transforming growth factor-beta induces renal epithelial jagged-1 expression in fibrotic disease. J Am Soc Nephrol 13:1499–1508CrossRefPubMed
Metadata
Title
Glomerular expression of transforming growth factor–β (TGF-β) isoforms in mice lacking CD2-associated protein
Authors
Robert P. Woroniecki
Mario Schiffer
Andrey S. Shaw
Frederick J. Kaskel
Erwin P. Bottinger
Publication date
01-03-2006
Publisher
Springer-Verlag
Published in
Pediatric Nephrology / Issue 3/2006
Print ISSN: 0931-041X
Electronic ISSN: 1432-198X
DOI
https://doi.org/10.1007/s00467-005-2102-7

Other articles of this Issue 3/2006

Pediatric Nephrology 3/2006 Go to the issue