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Analysis of WT1 gene expression during mouse nephrogenesis in organ culture

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Summary

The temporal and spatial expression patterns of the Wilms tumor gene, WT1, were studied during the organogenesis of the mouse kidneyin vitro. In situ hybridization and immunocytochemistry localized cellular expression of WT1 in whole kidney organ cultures to the induced metanephric mesenchyme and developing podocytes. Organ cultures were further characterized immunocytochemically with antibodies that specifically labeled the different tubular epithelial components and supporting mesenchyme of the developing nephrons. In organ cultures, the WT1 expression pattern could be visualized in induced metanephric mesenchyme and entire cell cohorts of differentiating podocytes. Expression of WT1 and cell specific markers were retained in short-term monolayer cultures of dissociated kidneys. The development of the metanephric kidneyin vitro involves a highly restricted temporal and spatial cellular expression pattern of WT1 which closely follows that observed in tissue sections from gestational kidney isolated during organogenesis in the mouse.

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References

  1. Armstrong, J. F.; Pritchard-Jones, K.; Bickmore, W. A., et al. The expression of the Wilms’ tumour gene, WT1, in the developing mammalian embryo. Mech. Dev. 40:85–97; 1992.

    Article  Google Scholar 

  2. Avner, E. D.; Sweeney, W. E. Polypeptide growth factors in metanephric growth and segmental nephron differentiation. Pediatr. Nephrol. 4: 372–377; 1990.

    Article  PubMed  CAS  Google Scholar 

  3. Avner, E. D.; Sweeney, W. E.; Finegold, D. N., et al. Growth factor requirements of metanephric differentiation in serum free murine organ culture. In Vitro Cell Dev. Biol. 21:297–304; 1985.

    Article  PubMed  CAS  Google Scholar 

  4. Bonetta, L.; Kuehn, S. E.; Huang, A., et al. Wilms tumor locus on 11p13 defined by multiple CpG Island-associated transcripts. Science 250:994–997; 1990.

    Article  PubMed  CAS  Google Scholar 

  5. Brenner, B. M. Determinants of epithelial differentiation during early nephrogenesis. J. Am. Soc. Nephrol. 1:127–139; 1990.

    PubMed  CAS  Google Scholar 

  6. Buckler, A. J.; Pelletier, J.; Haber, D. A., et al. The murine Wilms’ tumor gene (WT1): isolation, characterization, and expression during kidney development. Mol. Cell Biol. 11:1707–1712; 1991.

    PubMed  CAS  Google Scholar 

  7. Call, K. M.; Glasser, T.; Ito, C. Y., et al. Isolation and characterization of a zinc finger polypeptide gene at the human chromosome 11 Wilms’ tumor locus. Cell 60:509–520; 1990.

    Article  PubMed  CAS  Google Scholar 

  8. Chesa, P. G.; Rettig, W. J.; Thomson, T. M., et al. Immunohistochemical analysis of nerve growth factor receptor expression in normal and malignant human tissues. J. Histochem. Cytochem. 36:383–389; 1988.

    PubMed  CAS  Google Scholar 

  9. Chomczynski, P.; Sacchi, N. Single step method of RNA isolation by acid guanidinium-thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162:156–159; 1987.

    Article  PubMed  CAS  Google Scholar 

  10. Church, G. M.; Gilbert, W. Genomic sequencing. Proc. Natl. Acad. Sci. USA 81:1991–1995; 1984.

    Article  PubMed  CAS  Google Scholar 

  11. Coppes, M. J.; Campbell, C. E.; Williams, B. R. G. The role of WT1 in Wilms tumorigenesis. FASEB J. 7:886–895; 1993.

    PubMed  CAS  Google Scholar 

  12. Curto, K. A.; Sweeney, W. E.; Avner, E. D., et al. Immunocytochemical localization of ψ-glutamyltranspeptidase during fetal development of mouse kidney. J. Histochem. Cytochem. 36:159–166; 1988.

    PubMed  CAS  Google Scholar 

  13. Dey, B. R.; Sukhatme, V. P.; Roberts, A. B., et al. Repression of the transforming growth factor β1 gene by the Wilms tumor suppressor WT1 gene product. Mol. Endocrinol. 8:595–602; 1994.

    Article  PubMed  CAS  Google Scholar 

  14. Dressler, G. R.; Douglass, E. C. Pax-2 is a DNA-binding protein expressed in embryonic kidney and Wilms tumor. Proc. Natl. Acad. Sci. USA 89:1179–1183; 1992.

    Article  PubMed  CAS  Google Scholar 

  15. Dressler, G. R.; Deutsch, U.; Chowdhury, K., et al. Pax2, a new murine paired-box-containing gene and its expression in the developing excretory system. Development 109:787–795; 1990.

    PubMed  CAS  Google Scholar 

  16. Drummond, I. A.; Madden, S. L.; Rohives-Nutter, P., et al. Repression of the insulin-like growth factor II gene by the Wilms tumor suppressor WT1. Science 257:674–678; 1992.

    Article  PubMed  CAS  Google Scholar 

  17. Eccles, M. R.; Wallis, L. J.; Fidler, A. E., et al. Expression of the PAX2 gene in human fetal kidney and Wilms’ tumor. Cell Growth Differ. 3:279–289; 1992.

    PubMed  CAS  Google Scholar 

  18. Eccles, M. R.; Yun, K.; Reeve, A. E., et al. Comparative in situ hybridization analysis of PAX2, PAX8, and WT1 gene transcription in human fetal kidney and Wilms’ tumors. Am. J. Pathol. 146:40–45; 1995.

    PubMed  CAS  Google Scholar 

  19. Ekblom, P. Developmentally regulated conversion of mesenchyme to epithelium. FASEB J. 3:2141–2148; 1989

    PubMed  CAS  Google Scholar 

  20. Ekblom, P.; Thesleff, I. Role of transferrin and extracellular matrix components in kidney differentiation. Mod. Cell Biol. 4:85–128; 1985.

    CAS  Google Scholar 

  21. Ernfors, P.; Wetmore, C.; Eriksdotter-Nilsson, M., et al. The nerve growth factor receptor gene is expressed in both neuronal and non-neuronal tissues in the human fetus. Int. J. Dev. Neurosci. 9:57–66; 1991.

    Article  PubMed  CAS  Google Scholar 

  22. Ernfors, P.; Wetmore, C.; Olson, L., et al. Identification of cells in rat brain and peripheral tissues expressing mRNA for members of the nerve growth factor family. Neuron 5:511–526; 1990.

    Article  PubMed  CAS  Google Scholar 

  23. Fisher, D. A.; Salido, E. C.; Barajas, L. Epidermal growth factor and the kidney. Annu. Rev. Physiol. 51:61–80; 1989.

    Article  Google Scholar 

  24. Flenniken, A. M.; Williams, B. R. G. Developmental expression of the endogenous TIMP gene and a TIMP-lacZ fusion gene in transgenic mice. Genes Dev. 4:1094–1106; 1990.

    Article  PubMed  CAS  Google Scholar 

  25. Gessler, M.; Konig, A.; Bruns, G. A. P. The genomic organization and expression of the WT1 gene. Genomics 12:807–813; 1992.

    Article  PubMed  CAS  Google Scholar 

  26. Goldfarb, M. The fibroblast growth factor family. Cell Growth Differ. 1:439–445; 1990.

    PubMed  CAS  Google Scholar 

  27. Goodyer, P.; Dehbi, M.; Torban, E., et al. Repression of the retinoic acid receptor-α gene by the Wilms’ tumor suppressor gene, wt1. Oncogene 10:1125–1129; 1995.

    PubMed  CAS  Google Scholar 

  28. Gray, A.; Tam, A. W.; Dull, T. J., et al. Tissue-specific and developmentally regulated transcription of the insulin-like growth factor II gene. DNA 6:283–295; 1987.

    Article  PubMed  CAS  Google Scholar 

  29. Grobstein, C. Transfilter induction of tubules in mouse metanephrogenic mesenchyme. Exp. Cell Res. 10:424–440; 1956.

    Article  PubMed  CAS  Google Scholar 

  30. Hammerman, M. R.; O’Shea, M.; Muller, S. B. Role of growth factors in regulation of renal growth. Ann. Rev. Physiol. 55:305–321; 1993.

    Article  CAS  Google Scholar 

  31. Hogan, B.; Constantini, F.; Lacey, E., Section F. Visualizing genes and gene products. In: B. Hogan, R. Beddington, F. Constantini, E. Lacey (eds). Manipulating the mouse embryo. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press; 1986:234–235.

    Google Scholar 

  32. Huang, A.; Campbell, C. E.; Bonetta, L., et al. Tissue, developmental and tumor-specific expression of divergent transcripts in Wilms tumor Science 250:991–994; 1990.

    Article  PubMed  CAS  Google Scholar 

  33. Kistler, A.; Wessner, B. Extensive growth and differentiation of limb bud cells grown on polycarbonate membranes. In Vitro Cell Dev. Biol. 27:105–109; 1991.

    Article  Google Scholar 

  34. Kreidberg, J. A.; Sariola, H.; Loring, J. M., et al. WT-1 is required for early kidney development. Cell 74:679–691; 1993.

    Article  PubMed  CAS  Google Scholar 

  35. Leiderman, L. J.; Tucker, J. A.; Dennis, V. W. Growth and differentiation of opossum kidney cells on microscopically transparent microporous membranes. Tissue Cell 21:355–360; 1989.

    Article  PubMed  CAS  Google Scholar 

  36. Liu, L.; Dunn, S. T.; Christakos, S., et al. Calbindin-D28k gene expression in the developing mouse kidney. Kidney Int. 44:322–330; 1993.

    Article  PubMed  CAS  Google Scholar 

  37. Madden, S. L.; Cook, D. M.; Morris, J. F., et al. Transcriptional repression mediated by the WT1 Wilms tumor gene product. Science 253:1550–1553; 1991.

    Article  PubMed  CAS  Google Scholar 

  38. Madden, S. L.; Cook, D. M.; Rauscher, F. J. A structure-function analysis of transcriptional repression mediated by WT1, Wilms’ tumor suppressor protein. Oncogene 8:1713–1720; 1993.

    PubMed  CAS  Google Scholar 

  39. Mendley, S. R.; Toback, F. G. Autocrine and paracrine regulation of kidney epithelial cell growth. Annu. Rev. Physiol. 51:33–50; 1988.

    Article  Google Scholar 

  40. Mugrauer, G.; Ekblom, P. Contrasting expression patterns of three members of the myc family of protooncogenes in the developing and adult mouse kidney. J. Cell Biol. 112:13–25; 1991.

    Article  PubMed  CAS  Google Scholar 

  41. Park, S.; Schalling, M.; Bernard, A., et al. The Wilms tumour gene WT1 is expressed in murine mesoderm-derived tissues and mutated in mesothelioma. Nature Genet. 4:415–420; 1993.

    Article  PubMed  CAS  Google Scholar 

  42. Pelletier, J.; Schalling, M.; Buckler, A. J., et al. Expression of the Wilms’ tumor gene WT1 in the murine urogenital system. Genes Develop. 5:1345–1356; 1991.

    Article  PubMed  CAS  Google Scholar 

  43. Powers, A. C.; Rabizadeh, A.; Akeson, R., et al. Characterization of monoclonal antibody 3G5 and utilization of this antibody to immobilize pancreatic cell gangliosides in a solid phase radioassay. Endocrinology 114:1338–1343; 1984.

    Article  PubMed  CAS  Google Scholar 

  44. Pritchard-Jones, K.; Fleming, S. Cell types expressing the Wilms’ tumour gene (WT1) in Wilms’tumours: implication for tumour histogenesis. Oncogene 6:2211–2220; 1991.

    PubMed  CAS  Google Scholar 

  45. Pritchard-Jones, K.; Fleming, S.; Davidson, D., et al. The candidate Wilms’ tumor gene is involved in genitourinary development. Nature (Lond.) 346:194–197; 1990.

    Article  CAS  Google Scholar 

  46. Rackley, R. R.; Flenniken, A. M.; Kuriyan, N. P., et al. Expression of the Wilms’ tumor suppressor gene WT1 during mouse embryogenesis. Cell Growth Differ. 4:1–9; 1993.

    Google Scholar 

  47. Rauscher, F. J. The Wilms tumor gene product: a developmentally regulated transcription factor in the kidney that functions as a tumor suppressor. FASEB J. 7:896–903; 1993.

    PubMed  CAS  Google Scholar 

  48. Sariola, H.; Saarma, M.; Sainio, K., et al. Dependence of kidney morphogenesis on the expression of nerve growth factor receptor. Science 254:571–573; 1991.

    Article  PubMed  CAS  Google Scholar 

  49. Saxen, L. Organogenesis of the kidney. Cambridge University Press, Cambridge, (U.K.); 1987.

    Google Scholar 

  50. Segal, R.; Fine, L. G. Polypeptide growth factors and the kidney. Kidney Int. 36:S2-S10; 1989.

    Article  CAS  Google Scholar 

  51. Shamley, D. R.; Opperman, L. A.; Buffenstein, R., et al. Ontogeny of calbindin-D28K and calbindin-D9K in the mouse kidney, duodenum, cerebellum and placenta. Development 116:491–496; 1992.

    PubMed  CAS  Google Scholar 

  52. Sorokin, L.; Ekblom, P. Development of tubular and glomerular cells of the kidney. Kidney Int. 41:657–664; 1992.

    Article  PubMed  CAS  Google Scholar 

  53. Sweeney, W. E.; Avner, E. D. Intact organ culture of murine metanephros. J. Tissue Culture Methods 13:163–168; 1991.

    Article  Google Scholar 

  54. van Heyningen, V.; Bickmore, W. A.; Seawright, A., et al. Role for the Wilm’s tumor gene in genital development. Proc. Natl. Acad Sci. USA 87:5383–5386; 1990.

    Article  PubMed  Google Scholar 

  55. Wang, Z.-Y.; Madden, S. L.; Deuel, T. F., et al. The Wilms’ tumor gene product, WT1, represses transcription of the platelet-derived growth factor A-chain gene. J. Biol. Chem. 267:21999–22002; 1992.

    PubMed  CAS  Google Scholar 

  56. Wang, Z.-Y.; Qui, Q.-Q.; Enger, K. T., et al. A second transcriptionally active DNA-binding site for the Wilms tumor gene product, WT1. Proc. Natl. Acad. Sci. USA 90:8896–8900; 1993.

    Article  PubMed  CAS  Google Scholar 

  57. Weller, A.; Sorokin, L.; Illgen, E.-M., et al. Development and growth of mouse embryonic kidney in organ culture and modulation of development by soluble growth factor. Develop. Biol. 144:248–261; 1991.

    Article  PubMed  CAS  Google Scholar 

  58. Werner, H.; Shen-Orr, Z.; Rauscher, F. J., III, et al. Inhibition of cellular proliferation by the Wilms’ tumor suppressor WT1 is associated with suppression of insulin-like growth factor 1 receptor gene expression. Mol. Cell. Biol. 15:3516–3522; 1995.

    PubMed  CAS  Google Scholar 

  59. Werner, H.; Re, G. G.; Drummond, I. A., et al. Increased expression of the insulin-like growth factor I receptor gene, IGF1R, in Wilms tumor is correlated with the modulation of IGF1R promoter activity by the WT1 Wilms tumor gene product. Proc. Natl. Acad. Sci. USA 90:5828–5832; 1993.

    Article  PubMed  CAS  Google Scholar 

  60. Wheeler, E. F.; Bothwell, M. Spatiotemporal patterns of expression of NGF and the low-affinity NGF receptor in rat embryos suggest functional roles in tissue morphogenesis and myogenesis. J. Neurosci. 12:930–945; 1992.

    PubMed  CAS  Google Scholar 

  61. Yeger, H.; Cullinane, C.; Flenniken, A., et al. Coordinate expression of Wilms tumor genes correlates with Wilms tumor phenotypes. Cell Growth Differ. 3:855–865; 1992.

    PubMed  CAS  Google Scholar 

  62. Yeger, H.; Baumal, R.; Pawlin, G., et al. Phenotypic and molecular characterization of inducible human neuroblastoma cell lines. Differentiation 39:216–227; 1988.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Pathology, The Hospital for Sick Children, 555 University Avenue, M5G 1X8, Toronto, Ontario, Canada

    Herman Yeger, Diane Forget & Jennifer Alami

  2. Department of Cancer Biology, Cleveland Clinic Foundation Research Institute, 9500 Euclid Ave, 44195, Cleveland, Ohio

    Bryan R. G. Williams

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  1. Herman Yeger
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  2. Diane Forget
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  3. Jennifer Alami
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Yeger, H., Forget, D., Alami, J. et al. Analysis of WT1 gene expression during mouse nephrogenesis in organ culture. In Vitro Cell.Dev.Biol.-Animal 32, 496–504 (1996). https://doi.org/10.1007/BF02723053

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  • DOI: https://doi.org/10.1007/BF02723053

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