Skip to main content
PMC home page
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1993 Apr;142(4):1051–1060.

Targeting of the rasT24 oncogene to the proximal convoluted tubules in transgenic mice results in hyperplasia and polycystic kidneys.

D L Schaffner 1, R Barrios 1, C Massey 1, E I Bañez 1, C N Ou 1, S Rajagopalan 1, E Aguilar-Cordova 1, R M Lebovitz 1, P A Overbeek 1, M W Lieberman 1
PMCID: PMC1886875  PMID: 8097368

Abstract

Five families of transgenic mice were derived from one-cell-stage embryos injected with gamma GT-rasT24, a fusion gene consisting of the gamma-glutamyl transpeptidase (gamma GT) 5' flanking region containing promoter I linked to a mutated (codon 12) human H-ras oncogene. The transgene was expressed selectively in the kidneys, eyes, and brains of all families as determined by reverse transcription-polymerase chain reaction, nuclease protection assays, and in situ hybridization. In two of five families, kidney lesions consisting of proximal tubular hyperplasia, renal cysts, and microadenomas developed in male animals; males also expressed higher levels of gamma GT/rasT24 RNA. Early lesions consisted of proximal tubular hyperplasia as defined by alkaline phosphatase histochemistry, gamma GT immunohistochemistry, and electron microscopy and could be correlated with the presence of rasT24 RNA within the cystic proximal tubular epithelium by in situ hybridization. Advanced lesions also involved other segments of the nephron and consisted of cysts lined by a flattened unicellular layer of attenuated epithelium. No rasT24 could be identified within cystic lesions of the distal nephron and collecting tubules by in situ hybridization, and they most likely arise by external compression. Animals from the two transgenic strains exhibiting cystic lesions die of renal failure beginning at 8 months of age. No difference in cell-cycle parameters or DNA ploidy between transgenic and control kidneys was identified by flow cytometric analysis. No renal carcinomas developed. The primary renal effects of the H-rasT24 oncogene in this model system consist of proximal tubular hyperplasia and polycystic kidneys. This model appears to provide a useful in vivo system for the study of ras oncogene function and control of renal cell proliferation.

Full text

PDF
1055

Images in this article

1055Figure 1
on p.1055
1056Figure 2
on p.1056
1057Figure 3
on p.1057
1057Figure 4
on p.1057
1057Figure 5
on p.1057
1058Figure 6
on p.1058

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andres A. C., Schönenberger C. A., Groner B., Hennighausen L., LeMeur M., Gerlinger P. Ha-ras oncogene expression directed by a milk protein gene promoter: tissue specificity, hormonal regulation, and tumor induction in transgenic mice. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1299–1303. doi: 10.1073/pnas.84.5.1299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Avner E. D. Renal cystic disease. Insights from recent experimental investigations. Nephron. 1988;48(2):89–93. doi: 10.1159/000184884. [DOI] [PubMed] [Google Scholar]
  3. Bell J. C., Jardine K., McBurney M. W. Lineage-specific transformation after differentiation of multipotential murine stem cells containing a human oncogene. Mol Cell Biol. 1986 Feb;6(2):617–625. doi: 10.1128/mcb.6.2.617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bernstein J., Evan A. P., Gardner K. D., Jr Epithelial hyperplasia in human polycystic kidney diseases. Its role in pathogenesis and risk of neoplasia. Am J Pathol. 1987 Oct;129(1):92–101. [PMC free article] [PubMed] [Google Scholar]
  5. Bos J. L. ras oncogenes in human cancer: a review. Cancer Res. 1989 Sep 1;49(17):4682–4689. [PubMed] [Google Scholar]
  6. Capon D. J., Chen E. Y., Levinson A. D., Seeburg P. H., Goeddel D. V. Complete nucleotide sequences of the T24 human bladder carcinoma oncogene and its normal homologue. Nature. 1983 Mar 3;302(5903):33–37. doi: 10.1038/302033a0. [DOI] [PubMed] [Google Scholar]
  7. Chang E. H., Furth M. E., Scolnick E. M., Lowy D. R. Tumorigenic transformation of mammalian cells induced by a normal human gene homologous to the oncogene of Harvey murine sarcoma virus. Nature. 1982 Jun 10;297(5866):479–483. doi: 10.1038/297479a0. [DOI] [PubMed] [Google Scholar]
  8. Chobert M. N., Lahuna O., Lebargy F., Kurauchi O., Darbouy M., Bernaudin J. F., Guellaen G., Barouki R., Laperche Y. Tissue-specific expression of two gamma-glutamyl transpeptidase mRNAs with alternative 5' ends encoded by a single copy gene in the rat. J Biol Chem. 1990 Feb 5;265(4):2352–2357. [PubMed] [Google Scholar]
  9. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  10. DeBault L. E., Cancilla P. A. gamma-Glutamyl transpeptidase in isolated brain endothelial cells: induction by glial cells in vitro. Science. 1980 Feb 8;207(4431):653–655. doi: 10.1126/science.6101511. [DOI] [PubMed] [Google Scholar]
  11. Der C. J., Krontiris T. G., Cooper G. M. Transforming genes of human bladder and lung carcinoma cell lines are homologous to the ras genes of Harvey and Kirsten sarcoma viruses. Proc Natl Acad Sci U S A. 1982 Jun;79(11):3637–3640. doi: 10.1073/pnas.79.11.3637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Feramisco J. R., Gross M., Kamata T., Rosenberg M., Sweet R. W. Microinjection of the oncogene form of the human H-ras (T-24) protein results in rapid proliferation of quiescent cells. Cell. 1984 Aug;38(1):109–117. doi: 10.1016/0092-8674(84)90531-2. [DOI] [PubMed] [Google Scholar]
  13. Fujita J., Kraus M. H., Onoue H., Srivastava S. K., Ebi Y., Kitamura Y., Rhim J. S. Activated H-ras oncogenes in human kidney tumors. Cancer Res. 1988 Sep 15;48(18):5251–5255. [PubMed] [Google Scholar]
  14. Gardner K. D., Jr Pathogenesis of human cystic renal disease. Annu Rev Med. 1988;39:185–191. doi: 10.1146/annurev.me.39.020188.001153. [DOI] [PubMed] [Google Scholar]
  15. Gattone V. H., 2nd, Calvet J. P., Cowley B. D., Jr, Evan A. P., Shaver T. S., Helmstadter K., Grantham J. J. Autosomal recessive polycystic kidney disease in a murine model. A gross and microscopic description. Lab Invest. 1988 Aug;59(2):231–238. [PubMed] [Google Scholar]
  16. Godwin A. K., Lieberman M. W. Early and late responses to induction of rasT24 expression in Rat-1 cells. Oncogene. 1990 Aug;5(8):1231–1241. [PubMed] [Google Scholar]
  17. Grantham J. J., Geiser J. L., Evan A. P. Cyst formation and growth in autosomal dominant polycystic kidney disease. Kidney Int. 1987 May;31(5):1145–1152. doi: 10.1038/ki.1987.121. [DOI] [PubMed] [Google Scholar]
  18. Griffiths S. A., Manson M. M. Rat liver gamma glutamyl transpeptidase mRNA differs in the 5' untranslated sequence from the corresponding kidney mRNA. Cancer Lett. 1989 Jul 1;46(1):69–74. doi: 10.1016/0304-3835(89)90217-6. [DOI] [PubMed] [Google Scholar]
  19. Katsuki M., Kimura M., Hata J., Takahashi R., Nozawa S., Yokoyama M., Izawa M., Sekiya T., Nishimura S., Nomura T. Embryonal tumors from transgenic mouse zygotes carrying human activated c-Ha-ras genes. Mol Biol Med. 1989 Dec;6(6):567–572. [PubMed] [Google Scholar]
  20. Krishan A. Rapid flow cytofluorometric analysis of mammalian cell cycle by propidium iodide staining. J Cell Biol. 1975 Jul;66(1):188–193. doi: 10.1083/jcb.66.1.188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Land H., Parada L. F., Weinberg R. A. Tumorigenic conversion of primary embryo fibroblasts requires at least two cooperating oncogenes. Nature. 1983 Aug 18;304(5927):596–602. doi: 10.1038/304596a0. [DOI] [PubMed] [Google Scholar]
  22. Leder A., Kuo A., Cardiff R. D., Sinn E., Leder P. v-Ha-ras transgene abrogates the initiation step in mouse skin tumorigenesis: effects of phorbol esters and retinoic acid. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9178–9182. doi: 10.1073/pnas.87.23.9178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Li Y. C., Seyama T., Godwin A. K., Winokur T. S., Lebovitz R. M., Lieberman M. W. MTrasT24, a metallothionein-ras fusion gene, modulates expression in cultured rat liver cells of two genes associated with in vivo liver cancer. Proc Natl Acad Sci U S A. 1988 Jan;85(2):344–348. doi: 10.1073/pnas.85.2.344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lumpkin C. K., Knepper J. E., Butel J. S., Smith J. R., Pereira-Smith O. M. Mitogenic effects of the proto-oncogene and oncogene forms of c-H-ras DNA in human diploid fibroblasts. Mol Cell Biol. 1986 Aug;6(8):2990–2993. doi: 10.1128/mcb.6.8.2990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. MacKay K., Striker L. J., Pinkert C. A., Brinster R. L., Striker G. E. Glomerulosclerosis and renal cysts in mice transgenic for the early region of SV40. Kidney Int. 1987 Dec;32(6):827–837. doi: 10.1038/ki.1987.283. [DOI] [PubMed] [Google Scholar]
  26. McDivitt R. W., Stone K. R., Meyer J. S. A method for dissociation of viable human breast cancer cells that produces flow cytometric kinetic information similar to that obtained by thymidine labeling. Cancer Res. 1984 Jun;44(6):2628–2633. [PubMed] [Google Scholar]
  27. Miller K. G., Sollner-Webb B. Transcription of mouse rRNA genes by RNA polymerase I: in vitro and in vivo initiation and processing sites. Cell. 1981 Nov;27(1 Pt 2):165–174. doi: 10.1016/0092-8674(81)90370-6. [DOI] [PubMed] [Google Scholar]
  28. Nanus D. M., Ebrahim S. A., Bander N. H., Real F. X., Pfeffer L. M., Shapiro J. R., Albino A. P. Transformation of human kidney proximal tubule cells by ras-containing retroviruses. Implications for tumor progression. J Exp Med. 1989 Mar 1;169(3):953–972. doi: 10.1084/jem.169.3.953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Nanus D. M., Mentle I. R., Motzer R. J., Bander N. H., Albino A. P. Infrequent ras oncogene point mutations in renal cell carcinoma. J Urol. 1990 Jan;143(1):175–178. doi: 10.1016/s0022-5347(17)39905-6. [DOI] [PubMed] [Google Scholar]
  30. Ohgaki H., Kleihues P., Hard G. C. Ki-ras mutations in spontaneous and chemically induced renal tumors of the rat. Mol Carcinog. 1991;4(6):455–459. doi: 10.1002/mc.2940040608. [DOI] [PubMed] [Google Scholar]
  31. Overbeek P. A., Aguilar-Cordova E., Hanten G., Schaffner D. L., Patel P., Lebovitz R. M., Lieberman M. W. Coinjection strategy for visual identification of transgenic mice. Transgenic Res. 1991 Dec;1(1):31–37. doi: 10.1007/BF02512994. [DOI] [PubMed] [Google Scholar]
  32. Parada L. F., Tabin C. J., Shih C., Weinberg R. A. Human EJ bladder carcinoma oncogene is homologue of Harvey sarcoma virus ras gene. Nature. 1982 Jun 10;297(5866):474–478. doi: 10.1038/297474a0. [DOI] [PubMed] [Google Scholar]
  33. Preminger G. M., Koch W. E., Fried F. A., McFarland E., Murphy E. D., Mandell J. Murine congenital polycystic kidney disease: a model for studying development of cystic disease. J Urol. 1982 Mar;127(3):556–560. doi: 10.1016/s0022-5347(17)53911-7. [DOI] [PubMed] [Google Scholar]
  34. Quaife C. J., Pinkert C. A., Ornitz D. M., Palmiter R. D., Brinster R. L. Pancreatic neoplasia induced by ras expression in acinar cells of transgenic mice. Cell. 1987 Mar 27;48(6):1023–1034. doi: 10.1016/0092-8674(87)90710-0. [DOI] [PubMed] [Google Scholar]
  35. Rajagopalan S., Park J. H., Patel P. D., Lebovitz R. M., Lieberman M. W. Cloning and analysis of the rat gamma-glutamyltransferase gene. J Biol Chem. 1990 Jul 15;265(20):11721–11725. [PubMed] [Google Scholar]
  36. Rutenburg A. M., Kim H., Fischbein J. W., Hanker J. S., Wasserkrug H. L., Seligman A. M. Histochemical and ultrastructural demonstration of gamma-glutamyl transpeptidase activity. J Histochem Cytochem. 1969 Aug;17(8):517–526. doi: 10.1177/17.8.517. [DOI] [PubMed] [Google Scholar]
  37. Saitoh A., Kimura M., Takahashi R., Yokoyama M., Nomura T., Izawa M., Sekiya T., Nishimura S., Katsuki M. Most tumors in transgenic mice with human c-Ha-ras gene contained somatically activated transgenes. Oncogene. 1990 Aug;5(8):1195–1200. [PubMed] [Google Scholar]
  38. Sandgren E. P., Quaife C. J., Pinkert C. A., Palmiter R. D., Brinster R. L. Oncogene-induced liver neoplasia in transgenic mice. Oncogene. 1989 Jun;4(6):715–724. [PubMed] [Google Scholar]
  39. Shichi H., Mahalak S. M., Sakamoto S., Lin W. L., Essner E. S. Immunocytochemical localization of gamma-glutamyl transpeptidase in porcine ciliary epithelium. Exp Eye Res. 1991 Jul;53(1):39–46. doi: 10.1016/0014-4835(91)90142-2. [DOI] [PubMed] [Google Scholar]
  40. Sinn E., Muller W., Pattengale P., Tepler I., Wallace R., Leder P. Coexpression of MMTV/v-Ha-ras and MMTV/c-myc genes in transgenic mice: synergistic action of oncogenes in vivo. Cell. 1987 May 22;49(4):465–475. doi: 10.1016/0092-8674(87)90449-1. [DOI] [PubMed] [Google Scholar]
  41. Suda Y., Aizawa S., Hirai S., Inoue T., Furuta Y., Suzuki M., Hirohashi S., Ikawa Y. Driven by the same Ig enhancer and SV40 T promoter ras induced lung adenomatous tumors, myc induced pre-B cell lymphomas and SV40 large T gene a variety of tumors in transgenic mice. EMBO J. 1987 Dec 20;6(13):4055–4065. doi: 10.1002/j.1460-2075.1987.tb02751.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Tremblay P. J., Pothier F., Hoang T., Tremblay G., Brownstein S., Liszauer A., Jolicoeur P. Transgenic mice carrying the mouse mammary tumor virus ras fusion gene: distinct effects in various tissues. Mol Cell Biol. 1989 Feb;9(2):854–859. doi: 10.1128/mcb.9.2.854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Trudel M., D'Agati V., Costantini F. C-myc as an inducer of polycystic kidney disease in transgenic mice. Kidney Int. 1991 Apr;39(4):665–671. doi: 10.1038/ki.1991.80. [DOI] [PubMed] [Google Scholar]
  44. Yokoyama T., Silversides D. W., Waymire K. G., Kwon B. S., Takeuchi T., Overbeek P. A. Conserved cysteine to serine mutation in tyrosinase is responsible for the classical albino mutation in laboratory mice. Nucleic Acids Res. 1990 Dec 25;18(24):7293–7298. doi: 10.1093/nar/18.24.7293. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES