Detection of c-Sis Proto-Oncogene Transcripts by Direct Enzyme-Labeled cDNA Probes and in situ Hybridization

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 li...

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Bibliographic Details
Published in:In Vitro Cellular & Developmental Biology - Animal 1992-02, Vol.28A (2), p.102-108
Main Authors: MCCLINTOCK, J. T, CHAN, I.-J, THAKER, S. R, KATIAL, A, TAUB, F. E, AOTAKI-KEEN, A. E, HJELMELAND, L. M
Format: Article
Language:English
Subjects:
Biological and medical sciences
Cell Division
Cell growth
Cell Line
Cell lines
Complementary DNA
Diverse techniques
DNA Probes
Fundamental and applied biological sciences. Psychology
Gene Expression - drug effects
Humans
In situ hybridization
In Vitro Techniques
Messenger RNA
Molecular and cellular biology
Neuroglia
Nucleic Acid Hybridization
Platelet-Derived Growth Factor - genetics
Platelets
Proto oncogenes
RNA, Messenger - genetics
Signal amplification
Silver
Tetradecanoylphorbol Acetate - pharmacology
Thrombin - pharmacology
Transforming Growth Factor beta - pharmacology
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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 ($TGFB_1$), 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$TGFB_1-treated$RPE cells, with a portion of these cells displaying a hybridization response in the nuclear region. When compared to PMA- and thrombin-treated cells,$TGFB_1$stimulated the RPE cell line to yield the greatest amount of detectable PDGF mRNA. In addition, the hybridization response observed in$TGFB_1-treated$cells was shown to be RNA dependent.
ISSN:0883-8364
2327-431X
1543-706X
DOI:10.1007/BF02631013