Skip to main content

Advertisement

SpringerLink
Log in

Forced expression of vascular endothelial growth factor-A in podocytes decreases mesangial cell numbers and attenuates endothelial cell differentiation in the mouse glomerulus

  • Original article
  • Published:
Clinical and Experimental Nephrology Aims and scope Submit manuscript

Abstract

Background

Glomerular podocyte-derived vascular endothelial growth factor (VEGF) is indispensable for the migration and proliferation of glomerular endothelial cells. In contrast, podocyte-specific Vegf overexpression leads to the collapse of glomerular tufts; however, the mechanisms underlying this outcome have not yet been reported.

Methods

To further clarify the effects of elevated levels of Vegf expression on glomerular cells, we established a dual transgenic mouse line in which Vegf was exclusively and inducibly expressed in podocytes under the control of the “Tet-on system” (Podocin-rtTA/TetO-Vegf164 mice).

Results

Macroscopic and microscopic examination of Podocin-rtTA/TetO-Vegf164 animals following Vegf induction identified the presence of prominent red bloody spots. In addition, the endothelial cell number was increased along with enlargement of the subendothelial spaces. We also observed impaired endothelial fenestrations and aberrant plasmalemmal vesicle-associated protein-1 (PV-1) expression. In contrast, the mesangial cell number markedly decreased, resulting in a glomerular tuft intussusceptive splitting defect. Furthermore, whereas platelet-derived growth factor-B (PDGF-B) expression in the glomerular cells of Podocin-rtTA/TetO-Vegf164 mice was not decreased, phospho-PDGF receptor immunoreactivity in the mesangial cells was significantly decreased when compared to wild-type animals.

Conclusion

Taken together, the results of this study indicated that the upregulation of podocyte VEGF decreased the number of mesangial cells, likely owing to inhibition of PDGF-B-mediated signaling.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Carmeliet P, Ferreira V, Breier G, Pollefeyt S, Kieckens L, Gertsenstein M, Fahrig M, Vandenhoeck A, Harpal K, Eberhardt C, Declercq C, Pawling J, Moons L, Collen D, Risau W, Nagy A. Abnormal blood vessel development and lethality in embryos lacking a single VEGF allele. Nature. 1996;380:435–9.

    Article  CAS  PubMed  Google Scholar 

  2. Coultas L, Chawengsaksophak K, Rossant J. Endothelial cells and VEGF in vascular development. Nature. 2005;438:937–45.

    Article  CAS  PubMed  Google Scholar 

  3. Benjamin LE, Hemo I, Keshet E. A plasticity window for blood vessel remodelling is defined by pericyte coverage of the preformed endothelial network and is regulated by PDGF-B and VEGF. Development. 1998;125:1591–8.

    CAS  PubMed  Google Scholar 

  4. Senger DR, Galli SJ, Dvorak AM, Perruzzi CA, Harvey VS, Dvorak HF. Tumor cells secrete a vascular permeability factor that promotes accumulation of ascites fluid. Science. 1983;219:983–5.

    Article  CAS  PubMed  Google Scholar 

  5. Tufro A, Norwood VF, Carey RM, Gomez RA. Vascular endothelial growth factor induces nephrogenesis and vasculogenesis. J Am Soc Nephrol. 1999;10:2125–34.

    CAS  PubMed  Google Scholar 

  6. Eremina V, Sood M, Haigh J, Nagy A, Lajoie G, Ferrara N, Gerber HP, Kikkawa Y, Miner JH, Quaggin SE. Glomerular-specific alterations of VEGF-A expression lead to distinct congenital and acquired renal diseases. J Clin Invest. 2003;111:707–16.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Yang JC, Haworth L, Sherry RM, Hwu P, Schwartzentruber DJ, Topalian SL, Steinberg SM, Chen HX, Rosenberg SA. A randomized trial of bevacizumab, an anti-vascular endothelial growth factor antibody, for metastatic renal cancer. N Engl J Med. 2003;349:427–34.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Veron D, Reidy KJ, Bertuccio C, Teichman J, Villegas G, Jimenez J, Shen W, Kopp JB, Thomas DB, Tufro A. Overexpression of VEGF-A in podocytes of adult mice causes glomerular disease. Kidney Int. 2010;77:989–99.

    Article  CAS  PubMed  Google Scholar 

  9. Ueda H, Miyazaki Y, Matsusaka T, Utsunomiya Y, Kawamura T, Hosoya T, Ichikawa I. Bmp in podocytes is essential for normal glomerular capillary formation. J Am Soc Nephrol. 2008;19:685–94.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Shigehara T, Zaragoza C, Kitiyakara C, Takahashi H, Lu H, Moeller M, Holzman LB, Kopp JB. Inducible podocyte-specific gene expression in transgenic mice. J Am Soc Nephrol. 2003;14:1998–2003.

    CAS  PubMed  Google Scholar 

  11. Miyazaki Y, Shimizu A, Pastan I, Taguchi K, Naganuma E, Naganuma E, Suzuki T, Hosoya T, Yokoo T, Saito A, Miyata T, Yamamoto M, Matsusaka T. Keap1 inhibition attenuates glomerulosclerosis. Nephrol Dial Transplant. 2014;29:783–91.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Esser S, Wolburg K, Wolburg H, Breier G, Kurzchalia T, Risau W. Vascular endothelial growth factor induces endothelial fenestrations in vitro. J Cell Biol. 1998;140:947–59.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Ichimura K, Stan RV, Kurihara H, Sakai T. Glomerular endothelial cells form diaphragms during development and pathologic conditions. J Am Soc Nephrol. 2008;19:1463–71.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Toyoda M, Najafian B, Kim Y, Caramori ML, Mauer M. Podocyte detachment and reduced glomerular capillary endothelial fenestration in human type 1 diabetic nephropathy. Diabetes. 2007;56:2155–60.

    Article  CAS  PubMed  Google Scholar 

  15. Kikkawa Y, Virtanen I, Miner JH. Mesangial cells organize the glomerular capillaries by adhering to the G domain of laminin alpha5 in the glomerular basement membrane. J Cell Biol. 2003;161:187–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Lindahl P, Hellstrom M, Kalen M, Karlsson L, Karlsson L, Pekny M, Pekna M, Soriano P, Betsholtz C. Paracrine PDGF-B/PDGF-Rbeta signaling controls mesangial cell development in kidney glomeruli. Development. 1998;125:3313–22.

    CAS  PubMed  Google Scholar 

  17. Greenberg JI, Shields DJ, Barillas SG, Acevedo LM, Murphy E, Huang J, Scheppke L, Stockmann C, Johnson RS, Angle N, Cheresh DA. A role for VEGF as a negative regulator of pericyte function and vessel maturation. Nature. 2008;456:809–13.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Veron D, Reidy K, Marlier A, Bertuccio C, Villegas G, Jimenez J, Kashgarian M, Tufro A. Induction of podocyte VEGF164 overexpression at different stages of development causes congenital nephrosis or steroid-resistant nephrotic syndrome. Am J Pathol. 2010;177:2225–33.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Satchell SC, Braet F. Glomerular endothelial cell fenestrations: an integral component of the glomerular filtration barrier. Am J Physiol Renal Physiol. 2009;296:F947–56.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Sorensson J, Fierlbeck W, Heider T, Schwarz K, Park DS, Mundel P, Lisanti M, Ballermann BJ. Glomerular endothelial fenestrae in vivo are not formed from caveolae. J Am Soc Nephrol. 2002;13:2639–47.

    Article  PubMed  Google Scholar 

  21. Yamamoto I, Horita S, Takahashi T, Tanabe K, Fuchinoue S, Teraoka S, Hattori M, Yamaguchi Y. Glomerular expression of plasmalemmal vesicle-associated protein-1 in patients with transplant glomerulopathy. Am J Transplant. 2007;7:1954–60.

    Article  CAS  PubMed  Google Scholar 

  22. Schlondorff D. The glomerular mesangial cell: an expanding role for a specialized pericyte. FASEB J. 1987;1:272–81.

    Article  CAS  PubMed  Google Scholar 

  23. Lindahl P, Johansson BR, Leveen P, Betsholtz C. Pericyte loss and microaneurysm formation in PDGF-B-deficient mice. Science. 1997;277:242–5.

    Article  CAS  PubMed  Google Scholar 

  24. Leveen P, Pekny M, Gebre-Medhin S, Swolin B, Larsson E, Betsholtz C. Mice deficient for PDGF B show renal, cardiovascular, and hematological abnormalities. Genes Dev. 1994;8:1875–87.

    Article  CAS  PubMed  Google Scholar 

  25. Frank S, Stallmeyer B, Kampfer H, Schaffner C, Pfeilschifter J. Differential regulation of vascular endothelial growth factor and its receptor fms-like-tyrosine kinase is mediated by nitric oxide in rat renal mesangial cells. Biochem J. 1999;338(Pt 2):367–74.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Boyle SC, Liu Z, Kopan R. Notch signaling is required for the formation of mesangial cells from a stromal mesenchyme precursor during kidney development. Development. 2014;141:346–54.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This work was supported in part by a Grant-in-Aid for Scientific Research (C) (Grant number: 23591202 to Y. M.) from the Japanese Ministry of Education, Culture, Sports, Science, and Technology (MEXT). We thank Yamato Kikkawa (Tokyo University of Pharmacy and Life Sciences) for providing the anti-collagen IV antibodies and anti-laminin antibodies. We thank Mitsuko Tamatsukuri and Moeno Ishida for their excellent technical assistance.

Author information

Authors and Affiliations

  1. Division of Nephrology and Hypertension, Department of Internal Medicine, The Jikei University School of Medicine, 3-25-8 Nishi-shinbashi, Minato-ku, Tokyo, 105-8461, Japan

    Masahiro Suyama, Yoichi Miyazaki, Naoki Sugano, Hiroyuki Ueda, Tetsuya Kawamura, Makoto Ogura & Takashi Yokoo

  2. Department of Internal Medicine, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa, 259-1193, Japan

    Taiji Matsusaka

Authors
  1. Masahiro Suyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yoichi Miyazaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Taiji Matsusaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Naoki Sugano
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hiroyuki Ueda
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Tetsuya Kawamura
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Makoto Ogura
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Takashi Yokoo
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yoichi Miyazaki.

Ethics declarations

Conflict of interest

The authors have declared that no conflicts of interest exist.

Ethical approval

All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted (approval number: 24-004).

Informed consent

This section is not applicable to this study.

Electronic supplementary material

Below is the link to the electronic supplementary material.

10157_2017_1450_MOESM1_ESM.pdf

Supplementary material 1 (PDF 1533 kb) Supplementary Fig. 1. Histological comparison of kidney tissues between wild-type and Podocin-rtTA single transgenic mice at P0. (a–h) There were no appreciable differences between wild-type and Podocin-rtTA mice in staining with hematoxylin and eosin (a, b); anti-α-smooth muscle actin (αSMA), a marker for mesangial cells (c, d); CD31 (e, f), a marker for endothelial cells; and anti-Wilm’s tumor (WT-1), a marker for podocytes (g, h). (i–l) Paraformaldehyde-fixed paraffin-embedded specimens were reprocessed for the electron microscopy. Ultrastructures of the basement membrane (depicted by * in k, l), epithelial foot process (fp), and endothelial fenestration (black arrowhead) were intact in both mouse lines. Magnification: 200 × (a–h), scale bars = 10 μm in (i, j) and 500 nm in (k, l)

10157_2017_1450_MOESM2_ESM.pdf

Supplementary material 2 (PDF 369 kb) Supplementary Fig. 2. Component structures of the glomerular basement membrane from wild-type (WT) and Podocin-rtTA/TetO-Vegf164 (Tg) mice. Representative immunostaining for collagen IVα4 (a, b), laminin α5 (c, d), and laminin β2 (e, f) of WT (a, c, and e) and Tg (b, d, and f) mice at P0. Magnification: 400×. Rabbit anti-collagen IVα4 and rabbit anti-laminin α5 and β2 antibodies were kindly provided by Mr. Yamato Kikkawa (School of Pharmacy, Tokyo University of Pharmacy and Life Science, Tokyo, Japan) [15]

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Suyama, M., Miyazaki, Y., Matsusaka, T. et al. Forced expression of vascular endothelial growth factor-A in podocytes decreases mesangial cell numbers and attenuates endothelial cell differentiation in the mouse glomerulus. Clin Exp Nephrol 22, 266–274 (2018). https://doi.org/10.1007/s10157-017-1450-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10157-017-1450-5

Keywords

Search

Navigation