Skip to main content

Advertisement

SpringerLink
Log in

Podocyte-associated proteins FAT, alpha-actinin-4 and filtrin are expressed in Langerhans islets of the pancreas

  • Original Paper
  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Nephrin is a crucial podocyte molecule in the kidney glomerular filtration barrier and it is also expressed in Langerhans islet beta cells of the pancreas. Recently, genetic mapping of proteinuric kidney disease genes and animal models have revealed further important molecules for the kidney filtration function including alpha-actinin-4, podocin, FAT, and NEPH1. This study was addressed to explore the pancreatic expression of the podocyte molecules podocin, FAT, alpha-actinin-4, NEPH1, NEPH2, filtrin/NEPH3, synaptopodin and CD2 associated protein (CD2AP). The mRNA and protein expressions were studied by RT-PCR and immunoblotting, and localization in the pancreas was investigated by immunofluorescence. Of the nephrin-associated podocyte proteins, filtrin/NEPH3, FAT, and alpha-actinin-4 were found to be expressed in the pancreas at the gene and protein level and localized to Langerhans islets. Immunoreactivity with the podocin antibody was detected mostly in the exocrine pancreas. NEPH1 and synaptopodin expression was detected only at the mRNA level. Further studies are needed to unravel the functional role of these podocyte-associated molecules in the pancreatic Langerhans islets.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

CD2AP:

CD2-associated protein

cDNA:

complementary DNA

RT−:

reverse transcriptase negative

RT+:

reverse transcriptase positive

References

  1. Holthofer H, Ahola H, Solin ML, Wang S, Palmen T, Luimula P, Miettinen A, Kerjaschki D (1999) Nephrin localizes at the podocyte filtration slit area and is characteristically spliced in the human kidney. Am J Pathol 155:1681–1687

    PubMed  CAS  Google Scholar 

  2. Palmen T, Ahola H, Palgi J, Aaltonen P, Luimula P, Wang S, Jaakkola I, Knip M, Otonkoski T, Holthofer H (2001) Nephrin is expressed in the pancreatic beta cells. Diabetologia 44:1274–1280

    Article  PubMed  CAS  Google Scholar 

  3. Kestila M, Lenkkeri U, Mannikko M, Lamerdin J, McCready P, Putaala H, Ruotsalainen V, Morita T, Nissinen M, Herva R, Kashtan CE, Peltonen L, Holmberg C, Olsen A, Tryggvason K (1998) Positionally cloned gene for a novel glomerular protein––nephrin––is mutated in congenital nephrotic syndrome. Mol Cell 1:575–582

    Article  PubMed  CAS  Google Scholar 

  4. Holzman LB, St John PL, Kovari IA, Verma R, Holthofer H, Abrahamson DR (1999) Nephrin localizes to the slit pore of the glomerular epithelial cell. Kidney Int 56:1481–1491

    Article  PubMed  Google Scholar 

  5. Ruotsalainen V, Ljungberg P, Wartiovaara J, Lenkkeri U, Kestila M, Jalanko H, Holmberg C, Tryggvason K (1999) Nephrin is specifically located at the slit diaphragm of glomerular podocytes. Proc Natl Acad Sci USA 96:7962–7967

    Article  PubMed  CAS  Google Scholar 

  6. Tryggvason K (1999) Unraveling the mechanisms of glomerular ultrafiltration: nephrin, a key component of the slit diaphragm. J Am Soc Nephrol 10:2440–2445

    PubMed  CAS  Google Scholar 

  7. Putaala H, Soininen R, Kilpelainen P, Wartiovaara J, Tryggvason K (2001) The murine nephrin gene is specifically expressed in kidney, brain and pancreas: inactivation of the gene leads to massive proteinuria and neonatal death. Hum Mol Genet 10:1–8

    Article  PubMed  CAS  Google Scholar 

  8. Rantanen M, Palmen T, Patari A, Ahola H, Lehtonen S, Astrom E, Floss T, Vauti F, Wurst W, Ruiz P, Kerjaschki D, Holthofer H (2002) Nephrin TRAP mice lack slit diaphragms and show fibrotic glomeruli and cystic tubular lesions. J Am Soc Nephrol 13:1586–1594

    Article  PubMed  CAS  Google Scholar 

  9. Boute N, Gribouval O, Roselli S, Benessy F, Lee H, Fuchshuber A, Dahan K, Gubler MC, Niaudet P, Antignac C (2000) NPHS2, encoding the glomerular protein podocin, is mutated in autosomal recessive steroid-resistant nephrotic syndrome. Nat Genet 24:349–354

    Article  PubMed  CAS  Google Scholar 

  10. Ciani L, Patel A, Allen ND, Ffrench-Constant C (2003) Mice lacking the giant protocadherin mFAT1 exhibit renal slit junction abnormalities and a partially penetrant cyclopia and anophthalmia phenotype. Mol Cell Biol 23:3575–3582

    Article  PubMed  CAS  Google Scholar 

  11. Donoviel DB, Freed DD, Vogel H, Potter DG, Hawkins E, Barrish JP, Mathur BN, Turner CA, Geske R, Montgomery CA, Starbuck M, Brandt M, Gupta A, Ramirez-Solis R, Zambrowicz BP, et al. (2001) Proteinuria and perinatal lethality in mice lacking NEPH1, a novel protein with homology to NEPHRIN. Mol Cell Biol 21:4829–4836

    Article  PubMed  CAS  Google Scholar 

  12. Kaplan JM, Kim SH, North KN, Rennke H, Correia LA, Tong HQ, Mathis BJ, Rodriguez-Perez JC, Allen PG, Beggs AH, Pollak MR (2000) Mutations in ACTN4, encoding alpha-actinin-4, cause familial focal segmental glomerulosclerosis. Nat Genet 24:251–256

    Article  PubMed  CAS  Google Scholar 

  13. Kawachi H, Kurihara H, Topham PS, Brown D, Shia MA, Orikasa M, Shimizu F, Salant DJ (1997) Slit diaphragm-reactive nephritogenic MAb 5-1-6 alters expression of ZO-1 in rat podocytes. Am J Physiol 273:F984–F993

    PubMed  CAS  Google Scholar 

  14. Kos CH, Le TC, Sinha S, Henderson JM, Kim SH, Sugimoto H, Kalluri R, Gerszten RE, Pollak MR (2003) Mice deficient in alpha-actinin-4 have severe glomerular disease. J Clin Invest 111:1683–1690

    Article  PubMed  CAS  Google Scholar 

  15. Roselli S, Heidet L, Sich M, Henger A, Kretzler M, Gubler MC, Antignac C (2004) Early glomerular filtration defect and severe renal disease in podocin-deficient mice. Mol Cell Biol 24:550–560

    Article  PubMed  CAS  Google Scholar 

  16. 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 286:312–315

    Article  PubMed  CAS  Google Scholar 

  17. Schnabel E, Anderson JM, Farquhar MG (1990) The tight junction protein ZO-1 is concentrated along slit diaphragms of the glomerular epithelium. J Cell Biol 111:1255–1263

    Article  PubMed  CAS  Google Scholar 

  18. Kurihara H, Anderson JM, Farquhar MG (1992) Diversity among tight junctions in rat kidney: glomerular slit diaphragms and endothelial junctions express only one isoform of the tight junction protein ZO-1. Proc Natl Acad Sci USA 89:7075–7079

    Article  PubMed  CAS  Google Scholar 

  19. Reiser J, Kriz W, Kretzler M, Mundel P (2000) The glomerular slit diaphragm is a modified adherens junction. J Am Soc Nephrol 11:1–8

    PubMed  CAS  Google Scholar 

  20. Shih NY, Li J, Cotran R, Mundel P, Miner JH, Shaw AS (2001) CD2AP localizes to the slit diaphragm and binds to nephrin via a novel C-terminal domain. Am J Pathol 159:2303–2308

    PubMed  CAS  Google Scholar 

  21. Ihalmo P, Palmen T, Ahola H, Valtonen E, Holthofer H (2003) Filtrin is a novel member of nephrin-like proteins. Biochem Biophys Res Commun 300:364–370

    Article  PubMed  CAS  Google Scholar 

  22. Sellin L, Huber TB, Gerke P, Quack I, Pavenstadt H, Walz G (2003) NEPH1 defines a novel family of podocin interacting proteins. Faseb J 17:115–117

    PubMed  CAS  Google Scholar 

  23. Inoue T, Yaoita E, Kurihara H, Shimizu F, Sakai T, Kobayashi T, Ohshiro K, Kawachi H, Okada H, Suzuki H, Kihara I, Yamamoto T (2001) FAT is a component of glomerular slit diaphragms. Kidney Int 59:1003–1012

    Article  PubMed  CAS  Google Scholar 

  24. Ahola H, Heikkila E, Astrom E, Inagaki M, Izawa I, Pavenstadt H, Kerjaschki D, Holthofer H (2003) A novel protein, densin, expressed by glomerular podocytes. J Am Soc Nephrol 14:1731–1737

    Article  PubMed  CAS  Google Scholar 

  25. Mundel P, Gilbert P, Kriz W (1991) Podocytes in glomerulus of rat kidney express a characteristic 44 KD protein. J Histochem Cytochem 39:1047–1056

    PubMed  CAS  Google Scholar 

  26. Mundel P, Heid HW, Mundel TM, Kruger M, Reiser J, Kriz W (1997) Synaptopodin: an actin-associated protein in telencephalic dendrites and renal podocytes. J Cell Biol 139:193–204

    Article  PubMed  CAS  Google Scholar 

  27. Kawachi H, Koike H, Kurihara H, Yaoita E, Orikasa M, Shia MA, Sakai T, Yamamoto T, Salant DJ, Shimizu F (2000) Cloning of rat nephrin: expression in developing glomeruli and in proteinuric states. Kidney Int 57:1949–1961

    Article  PubMed  CAS  Google Scholar 

  28. Khoshnoodi J, Tryggvason K (2001) Unraveling the molecular make-up of the glomerular podocyte slit diaphragm. Exp Nephrol 9:355–359

    Article  PubMed  CAS  Google Scholar 

  29. Smoyer WE, Mundel P (1998) Regulation of podocyte structure during the development of nephrotic syndrome. J Mol Med 76:172–183

    Article  PubMed  CAS  Google Scholar 

  30. Somlo S, Mundel P (2000) Getting a foothold in nephrotic syndrome. Nat Genet 24:333–335

    Article  PubMed  CAS  Google Scholar 

  31. Tryggvason K, Ruotsalainen V, Wartiovaara J (1999) Discovery of the congenital nephrotic syndrome gene discloses the structure of the mysterious molecular sieve of the kidney. Int J Dev Biol 43:445–451

    PubMed  CAS  Google Scholar 

  32. Huber TB, Kottgen M, Schilling B, Walz G, Benzing T (2001) Interaction with podocin facilitates nephrin signaling. J Biol Chem 276:41543–41546

    Article  PubMed  CAS  Google Scholar 

  33. Wheeler DL, Church DM, Federhen S, Lash AE, Madden TL, Pontius JU, Schuler GD, Schriml LM, Sequeira E, Tatusova TA, Wagner L (2003) Database resources of the National Center for Biotechnology. Nucleic Acids Res 31:28–33

    Article  PubMed  CAS  Google Scholar 

  34. Rozen S, Skaletsky H (2000) Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol 132:365–386

    PubMed  CAS  Google Scholar 

  35. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410

    Article  PubMed  CAS  Google Scholar 

  36. Benson DA, Karsch-Mizrachi I, Lipman DJ, Ostell J, Rapp BA, Wheeler DL (2000) GenBank. Nucleic Acids Res 28:15–18

    Article  PubMed  CAS  Google Scholar 

  37. Kerjaschki D, Farquhar MG (1983) Immunocytochemical localization of the Heymann nephritis antigen (GP330) in glomerular epithelial cells of normal Lewis rats. J Exp Med 157:667–686

    Article  PubMed  CAS  Google Scholar 

  38. Honda K, Yamada T, Endo R, Ino Y, Gotoh M, Tsuda H, Yamada Y, Chiba H, Hirohashi S (1998) Actinin-4, a novel actin-bundling protein associated with cell motility and cancer invasion. J Cell Biol 140:1383–1393

    Article  PubMed  CAS  Google Scholar 

  39. Liu G, Kaw B, Kurfis J, Rahmanuddin S, Kanwar YS, Chugh SS (2003) Neph1 and nephrin interaction in the slit diaphragm is an important determinant of glomerular permeability. J Clin Invest 112:209–221

    Article  PubMed  CAS  Google Scholar 

  40. Roselli S, Gribouval O, Boute N, Sich M, Benessy F, Attie T, Gubler MC, Antignac C (2002) Podocin localizes in the kidney to the slit diaphragm area. Am J Pathol 160:131–139

    PubMed  CAS  Google Scholar 

  41. Asanuma K, Kim K, Oh J, Giardino L, Chabanis S, Faul C, Reiser J, Mundel P (2005) Synaptopodin regulates the actin-bundling activity of alpha-actinin in an isoform-specific manner. J Clin Invest 115:1188–1198

    Article  PubMed  CAS  Google Scholar 

  42. Dunne J, Hanby AM, Poulsom R, Jones TA, Sheer D, Chin WG, Da SM, Zhao Q, Beverley PC, Owen MJ (1995) Molecular cloning and tissue expression of FAT, the human homologue of the Drosophila fat gene that is located on chromosome 4q34-q35 and encodes a putative adhesion molecule. Genomics 30:207–223

    Article  PubMed  CAS  Google Scholar 

  43. Zanone MM, Favaro E, Doublier S, Lozanoska-Ochser B, Deregibus MC, Greening J, Huang GC, Klein N, Cavallo Perin P, Peakman M, Camussi G (2005) Expression of nephrin by human pancreatic islet endothelial cells. Diabetologia 48:1789–1797

    Article  PubMed  CAS  Google Scholar 

  44. Kuusniemi AM, Kestila M, Patrakka J, Lahdenkari AT, Ruotsalainen V, Holmberg C, Karikoski R, Salonen R, Tryggvason K, Jalanko H (2004) Tissue expression of nephrin in human and pig. Pediatr Res 55:774–781

    Article  PubMed  CAS  Google Scholar 

  45. Sun C, Kilburn D, Lukashin A, Crowell T, Gardner H, Brundiers R, Diefenbach B, Carulli JP (2003) Kirrel2, a novel immunoglobulin superfamily gene expressed primarily in beta cells of the pancreatic islets. Genomics 82:130–142

    Article  PubMed  CAS  Google Scholar 

  46. Collares-Buzato CB, Carvalho CP, Furtado AG, Boschero AC (2004) Upregulation of the expression of tight and adherens junction-associated proteins during maturation of neonatal pancreatic islets in vitro. J Mol Histol 35:811–822

    Article  PubMed  CAS  Google Scholar 

  47. Fallon MB, Gorelick FS, Anderson JM, Mennone A, Saluja A, Steer ML (1995) Effect of cerulein hyperstimulation on the paracellular barrier of rat exocrine pancreas. Gastroenterology 108:1863–1872

    Article  PubMed  CAS  Google Scholar 

  48. 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 108:1621–1629

    Article  PubMed  CAS  Google Scholar 

  49. Moeller MJ, Sanden SK, Soofi A, Wiggins RC, Holzman LB (2002) Two gene fragments that direct podocyte-specific expression in transgenic mice. J Am Soc Nephrol 13:1561–1567

    Article  PubMed  CAS  Google Scholar 

  50. Rinta-Valkama J, Aaltonen P, Lassila M, Palmen T, Tossavainen P, Knip M, Holthofer JH: Densin and filtrin in the pancreas and in the kidney, targets for humoral autoimmunity in patients with type 1 diabetes. Diabetes Metab Res Rev, 2006, (in press)

Download references

Acknowledgments

We thank Dr. Antignac, Dr. Huber, Dr. Inoue, Dr. Izawa, and Dr. Yamada for providing anti-podocin, anti-NEPH1, anti-FAT, anti-densin, and anti-alpha-actinin-4 antibodies, respectively. We also thank Ms Ritva Majuri for her technical assistance. The Finnish Kidney Foundation, the Finnish Diabetes Foundation, the Finnish Cultural Foundation, the Paavo Nurmi Foundation, and the European Union (LSHB-CT-2003–503364) have supported this study.

Author information

Authors and Affiliations

  1. Research Program in Molecular Medicine, Biomedicum Helsinki, Department of Bacteriology and Immunology, Haartman Institute, University of Helsinki, PB 63, 00014, Helsinki, Finland

    Johanna Rinta-Valkama, Tuula Palmén, Markus Lassila & Harry Holthöfer

  2. Helsinki University Central Hospital, Helsinki, Finland

    Johanna Rinta-Valkama, Tuula Palmén, Markus Lassila & Harry Holthöfer

Authors
  1. Johanna Rinta-Valkama
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tuula Palmén
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Markus Lassila
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Harry Holthöfer
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Harry Holthöfer.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rinta-Valkama, J., Palmén, T., Lassila, M. et al. Podocyte-associated proteins FAT, alpha-actinin-4 and filtrin are expressed in Langerhans islets of the pancreas. Mol Cell Biochem 294, 117–125 (2007). https://doi.org/10.1007/s11010-006-9251-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-006-9251-2

Keywords

Search

Navigation