In Vitro Cellular & Developmental Biology - Animal

, Volume 28, Issue 2, pp 102–108

Detection of c-Sis proto-oncogene transcripts by direct enzyme-labeled cDNA probes and in situ hybridization

Authors

  • J. T. McClintock
    • U.S. EPA, OPP, HED, SACB
    • Digene Diagnostics, Inc.
  • I-J. Chan
    • Digene Diagnostics, Inc.
  • S. R. Thaker
    • Digene Diagnostics, Inc.
  • A. Katial
    • Digene Diagnostics, Inc.
    • Department of OphthalmologyUniversity of California-Davis, Medical School
  • F. E. Taub
    • Digene Diagnostics, Inc.
    • Department of OphthalmologyUniversity of California-Davis, Medical School
  • A. E. Aotaki-Keen
    • Digene Diagnostics, Inc.
    • Department of OphthalmologyUniversity of California-Davis, Medical School
  • L. M. Hjelmeland
    • Digene Diagnostics, Inc.
    • Department of OphthalmologyUniversity of California-Davis, Medical School
Regular Papers

DOI: 10.1007/BF02631013

Cite this article as:
McClintock, J.T., Chan, I., Thaker, S.R. et al. In Vitro Cell Dev Biol - Animal (1992) 28: 102. doi:10.1007/BF02631013

Summary

Using in situ hybridization and platelet-derived growth factor (PDGF) cDNA probes labeled with horseradish peroxidase, PDGF-A and -B (c-cis proto-oncogene) mRNA transcripts were identified and localized in proliferating cultures. A human retinal pigment epithelial (RPE) cell line and a glial cell line were treated with either transforming growth factor beta-1 (TGFB1), phorbol-12-myristate-13-acetate (PMA), or thrombin from human plasma and compared for their ability to stimulate the production of PDGF-A and -B. Expression of both PDGF-A and -B transcripts were found to be localized predominantly in the cytoplasm of TGFB1-treated RPE cells, with a portion of these cells displaying a hybridization response in the nuclear region. When compared to PMA- and thrombin-treated cells, TGFB1 stimulated the RPE cell line to yield the greatest amount of detectable PDGF mRNA. In addition, the hybridization response observed in TGFB1-treated cells was shown to be RNA dependent.

Key words

c-sis proto-oncogenemRNA detectionhorseradish peroxidase-labeled cDNA probesin situ hybridization
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Copyright information

© Tissue Culture Association 1992