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

  • J. T. McClintock
  • I-J. Chan
  • S. R. Thaker
  • A. Katial
  • F. E. Taub
  • A. E. Aotaki-Keen
  • L. M. Hjelmeland
Regular Papers

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-oncogene mRNA detection horseradish peroxidase-labeled cDNA probes in situ hybridization 

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Copyright information

© Tissue Culture Association 1992

Authors and Affiliations

  • J. T. McClintock
    • 1
    • 2
  • I-J. Chan
    • 2
  • S. R. Thaker
    • 2
  • A. Katial
    • 2
    • 3
  • F. E. Taub
    • 2
    • 3
  • A. E. Aotaki-Keen
    • 2
    • 3
  • L. M. Hjelmeland
    • 2
    • 3
  1. 1.U.S. EPA, OPP, HED, SACBWashington, DC
  2. 2.Digene Diagnostics, Inc.Silver Spring
  3. 3.Department of OphthalmologyUniversity of California-Davis, Medical SchoolSacramento

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