Transcription of Melanogenesis Enzymes in Melanocytes: Dependence upon Culture Conditions and Co-Cultivation With Keratinocytes

Authors

  • STEFAN KIPPENBERGER,

    1. Zentrum der Dermatologie und Venerologie, Abteilung 1, Klinikum der Johann Wolfgang Goethe-Universität, Frankfurt/Main, Germany and
    2. AK Kinematische Zellforschung, Zoologisches Institut, Johann Wolfgang Goethe-Universität, Frankfurt/Main, Germany
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  • AUGUST BERND,

    Corresponding author
    1. Zentrum der Dermatologie und Venerologie, Abteilung 1, Klinikum der Johann Wolfgang Goethe-Universität, Frankfurt/Main, Germany and
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  • JÜRGEN BEREITER-HAHN,

    1. AK Kinematische Zellforschung, Zoologisches Institut, Johann Wolfgang Goethe-Universität, Frankfurt/Main, Germany
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  • ANA RAMIREZ-BOSCA,

    1. Zentrum der Dermatologie und Venerologie, Abteilung 1, Klinikum der Johann Wolfgang Goethe-Universität, Frankfurt/Main, Germany and
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  • ROLAND KAUFMANN,

    1. Zentrum der Dermatologie und Venerologie, Abteilung 1, Klinikum der Johann Wolfgang Goethe-Universität, Frankfurt/Main, Germany and
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  • HANS HOLZMANN

    1. Zentrum der Dermatologie und Venerologie, Abteilung 1, Klinikum der Johann Wolfgang Goethe-Universität, Frankfurt/Main, Germany and
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Address reprint requests to Dr. A. Bernd, Abteilung für Dermatologie 1, Zentrum der Dermatologie und Venerologie, Klinikum der J.W. Goethe Universität, Theodor-Stern-Kai 7, 60590 Frankfurt/Main, Germany.

Abstract

Eumelanogenesis of human skin melanocytes requires at least three enzymes: tyrosinase, TRP 1, and TRP 2. The regulation of these enzymes on transcriptional level was detected in a semiquantitative attempt. The total RNA of melanocytes was reverse-transcripted and followed by a PCR with degenerated primers for all three enzymes. The amplification products were related to each other densitometrically. We examined five different culture conditions: 1) melanocytes in a popular phorbolester containing F-10-medium, 2) melanocytes in a co-culture medium with EGF, 3) melanocytes in a co-culture medium with high calcium, 4) melanocytes co-cultured with keratinocytes in EGF containing co-culture medium, and 5) melanocytes co-cultured with keratinocytes in co-culture medium with high calcium. Melanocytes cultured in phorbolester containing F-10-medium featured transcripts of tyrosinase, TRP 1, and TRP 2 in the ratio 45:45:10. The same results were obtained for melanocytes co-cultured with keratinocytes under the two different culture conditions. In melanocytes cultured alone in co-culture media only TRP 1-transcripts were present. It is likely that under co-culture conditions a keratinocyte-derived factor supports the transcription of all three enzymes. For melanocytes in the phorbolester-containing melanocyte medium a proteinkinase C dependent regulation of transcription seems possible.

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