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Molecular and Cellular Biochemistry

, Volume 162, Issue 1, pp 65–73 | Cite as

Cyclic AMP regulates basement membrane heparan sulfate proteoglycan, perlecan, metabolism in rat glomerular epithelial cells

  • Cheol W. Ko
  • Basant Bhandari
  • J. Yee
  • W. C. Terhune
  • R. Maldonado
  • B. S. Kasinath
Article

Abstract

Perlecan, the basement membrane heparan sulfate proteoglycan (HSPG), has been fully cloned from mouse and human tissues. When a cRNA probe of murine perlecan cDNA was employed in RNase protection assay to test whether rat glomerular epithelial cells (GEC) constitutively express perlecan, several bands of hybridization were seen, suggesting that sequences between rat and murine perlecan may not be identical. Using primers based on published cDNA sequences of murine and human perlecan and polyA+ RNA of rat GEC, we synthesized a 497 by product (RPD-1) by RT PCR. The deduced aminoacid sequence showed an 85% and 88% homology with domain I of murine and human perlecan, respectively. The three putative sites containing the consensus sequence SGD for attachment of heparan sulfate chains were fully conserved in the rat perlecan as was a site (NFT) for attachment of N-linked oligosaccharide. RPD-I detected a > 9.5 kb transcript of perlecan in RNA of GEC, similar in size to that present in rat glomeruli. Employing a riboprobe synthesized from RPD-I in RNase protection assay we examined whether dbcAMP regulated perlecan expression in the GEC. At 1, 6, 24 and 48 h of incubation, l mM dbcAMP caused 43%, 32%, 47% and 40% reduction in mRNA abundance of perlecan, respectively. Immunoprecipitation showed a corresponding reduction of 61%, 70% and 65% in the synthesis of 35SO4 labeled basement membrane HSPG by the GEC following 12, 24 and 48 h of incubation with dbcAMP Following incubation for 1 and 24 h prostaglandins, PGE1 and PGE2 (1 uM), known activators of glomerular adenylate cyclase, reduced perlecan mRNA abundance to a similar extent as dbcAMP on northern analysis. Our results show that glomerular basement membrane HSPG synthesized by the GEC belongs to the perlecan family. Decrease of GEC perlecan gene expression and synthesis by cAMP and prostaglandins may be of relevance to proteinuric states characterized by activation of these mediators.

Key words

cAMP prostaglandins glomerular basement membrane perlecan glomerular epithelial cells 

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References

  1. 1.
    Rohrbach DH, Timpi R: In: D.H. Rohrbach and R. Timpl (eds). Molecular and Cellular Aspects of Basement Membranes. Academic Press, New York, 1993, pp 17–19Google Scholar
  2. 2.
    Kasinath BS, Kanwar YS: Glomerular basement membrane: Biology and physiology, In: D.H. Rohrbach and R. Timpl (eds). Molecular and Cellular Aspects of Basement Membranes. Academic Press, New York, 1993, pp 89–1206Google Scholar
  3. 3.
    Kanwar YS, Linker A, Farquhar MG: Increased permeability of the basement membrane to ferritin after removal of glycosaminoglycans (heparan sulfate) by enzymatic digestion. J Cell Biol 86: 688–693, 1980Google Scholar
  4. 4.
    Rosenzweig LJ, Kanwar YS: Removal of sulfated (heparan sulfate) and nonsulfated (hyaluronic acid) glycosaminoglycans result in increased permeability of the glomerular basement membrane to 125I-bovine serum albumin. Lab Invest 47: 177–184, 1982Google Scholar
  5. 5.
    Vernier RL, Steffes MW, Sisson-Ross S, Mauer SM: Heparan sulfate proteoglycan in the glomerular basement membrane of type I diabetes mellitus. Kidney Int 1070–1080, 1992Google Scholar
  6. 6.
    van den Born J, Lambert PWJ, van den Heuvel, Bakker MAH, Veerkamp JH, Assmann KJM, Weening JJ, Berden JoHM: Distribution of GBM heparan sulfate proteoglycan core protein and side chains in glomerular diseases. Kidney Int 43: 454–463, 1993Google Scholar
  7. 7.
    Kasinath BS, Block JA, Singh AK, Terhune WC, Maldonado R, Davalath S, Wanna L: Regulation of rat glomerular epithelial cell proteoglycans by high glucose medium. Arch Biochem Biophys 309: 149–159, 1994Google Scholar
  8. 8.
    Kasinath BS, Grellier P, Ghosh-Choudhury G, Abboud SL: Regulation of basement membrane heparan sulfate proteoglycan, perlecan, gene expression in glomerular epithelial cells by high glucose medium. J Cellular Physiol 167: 131–136, 1996Google Scholar
  9. 9.
    Noonan DM, Fulle A, Valente P, Cai S, Horigan E, Sasaki M, Yamade Y, Hassell JR: The complete seauence of perlecan, a basement membrane heparan sulfate proteoglycan, reveals extensive similarity to Laminin A chain, low density lipoprotein receptor and the neural cell adhesion molecule. J Biol Chem 266: 22939–22947, 1991Google Scholar
  10. 10.
    Kallunki P, Tryggvason K: Human basement membrane heparan sulfate proteoglycan core protein: A 467 kd protein containing multiple domains resembling elements of the low density lipoprotein receptor, laminin, neural cell adhesion molecule and epidermal growth factor. J Cell Biol 116: 559–571, 1992Google Scholar
  11. 11.
    Kasinath BS, Singh AK, Kanwar YS, Lewis EJ: Effect of puromycin aminonucleoside on HSPG core protein content of glomerular epithelial cells. Am J Physiol 255: F590-F596, 1988Google Scholar
  12. 12.
    Kasinath BS, Singh AK, Kanwar YS, Lewis EJ: Dexamethasone increases heparan sulfate proteoglycan core protein content of glomerular epithelial cells. J Lab Clin Med 115: 196–20, 1990Google Scholar
  13. 13.
    Sing AK, Kasinath BS: Metabolic fate of monavalent and multivalent antibodies of Heymann nephritis following formation of surface immune complexes on glomerular epithelial cells. Clin Exp Immun 94: 403–411, 1993Google Scholar
  14. 14.
    Kasinath BS: Effect of insulin on high glucose medium induced changes in rat glomerular epithelial cell metabolism of glycoconjugates. Arch Biochem Biophys 318: 286–294, 1995.Google Scholar
  15. 15.
    Chomczynski P, Sacchi N: Single step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159, 1987CrossRefPubMedGoogle Scholar
  16. 16.
    Bassett P, Bellocq JP, Worlf C, Stoll I, Hutin P, Limacher JM, Podhajcer OL, Chenard MP, Rio MC, Chambon P: A novel metalloproteinase gene specifically expressed in stromal cells of breast carcinoma. Nature 348: 699–704, 1993Google Scholar
  17. 17.
    Ledbetter SR, Tyree B, Hassell JR, Horigan EA: Identification of precursor protein to basement membrane heparan sulfate proteoglycan. J Biol Chem 260: 8106–8113, 1985Google Scholar
  18. 18.
    Makino H, Gibbons JT, Reddy MK, Kanwar YS: Nephritogenicity of antibodies to proteoglycans of glomerular basement membrane-I. J Clin Invest 77: 142–156, 1986Google Scholar
  19. 19.
    Fukui M, Nakamura T, Ebihara I, Shirato I, Tomino Y, Koide H: ECM gene expression and its modulation by insulin in diabetic rats. Diabetes 41: 1520–1527, 1992Google Scholar
  20. 20.
    Dworkin ZD, Ichikawa I, Brenner BM: Hormonal modulation of glomerular function. Am J Physiol 244: F95–104, 1983Google Scholar
  21. 21.
    Klein DJ, Brown DM, Oegema RT Jr., Brenchley PE, Anderson JC, Dickinson MAJ, Horigan EA, Hassell JR: Glomerular basement membrane proteoglycans are derived from a large precursor. J Cell Biol 1106: 963–970, 1988Google Scholar
  22. 22.
    Kokenyesi R, Silbert JE: Formation of heparan sulfate or chondroitin/dennatan sulfate on recombinant domain I of mouse perlecan expressed in Chinese hamster ovary cells. Biochem Biophys Res Commun 211: 262–267, 1995Google Scholar
  23. 23.
    Aviezer D, Hecht D, Safran, Eisinger M, David G, Yayon A: Perlecan, basal lamina proteoglycan, promotes basic fibroblast growth factorreceptor binding, mitogenesis and angiogenesis. Cell 79: 1005–1013, 1994Google Scholar
  24. 24.
    Zhang L, Esko JD: Aminoacid determinants that drive heparan sulfate assembly in a proteoglycan. J Biol Chem 269: 19295–19229, 1994Google Scholar
  25. 25.
    Chakravarti S, Hassell JR, Phillips SL: Perlecan gene expression precedes laminin gene expression during differentiation of F-9 embryonal carcinoma cells. Dev Dynamics 197: 107–114, 1993Google Scholar
  26. 26.
    Cohen I, Grassel S, Murdoch AD, Iozzo RV: Structural characterization of the complete human perlecan gene and its promoter. Proc Natl Acad Sci USA 90: 10104–10408, 1993Google Scholar
  27. 27.
    Yeaman C, Rapraeger AC: Post-transcriptional regulation of syndecanI expression by cAMP in peritoneal macrophages. J Cell Biol 122: 941–950, 1993Google Scholar
  28. 28.
    Lalli E, Sassone-Corsi P: Signal transduction and gene regulation: The nuclear response to cAMP. J Biol Chem 269: 17359–17362, 1994Google Scholar
  29. 29.
    Hagiwara M, Alberts A, Brindle P, Meinkoth J, Peramisco J, Deng T, Karin M, Shenolikar S, Momminy M: Transcriptional attenuation following cAMP induction requires PP-1 mediated dephosphorylation of CREB. Cell 70: 105–114, 1992Google Scholar
  30. 30.
    Alavi N, Lianos EA, Venuto RC, Mookerjee BK, Bentzel CJ: Reduction of proteinuria by indomethacin in patients with nephrotic syndrome. Amer J Kidney Dis 8: 397–403, 1986Google Scholar
  31. 31.
    Dunn MJ: Prostaglandins, Angiotensin II and proteinuria. Nephron 55(Supp 1): 30–37, 1990Google Scholar

Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • Cheol W. Ko
    • 1
  • Basant Bhandari
    • 1
  • J. Yee
    • 1
  • W. C. Terhune
    • 1
  • R. Maldonado
    • 1
  • B. S. Kasinath
    • 1
  1. 1.Department of MedicineUniversity of Texas Health Science Center, Audie L. Murphy Memorial Veterans' Administration HospitalSan AntonioUSA

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