Transcription of melanogenesis enzymes in melanocytes: dependence upon culture conditions and co-cultivation with keratinocytes

Pigment Cell Res. 1996 Aug;9(4):179-84. doi: 10.1111/j.1600-0749.1996.tb00107.x.

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.

MeSH terms

  • Coculture Techniques
  • Culture Media
  • Humans
  • Intramolecular Oxidoreductases*
  • Isomerases / genetics*
  • Keratinocytes
  • Melanocytes / enzymology*
  • Membrane Glycoproteins*
  • Monophenol Monooxygenase / genetics*
  • Oxidoreductases*
  • Polymerase Chain Reaction
  • Proteins / genetics*
  • Skin / embryology
  • Skin / enzymology
  • Skin / pathology
  • Transcription, Genetic*

Substances

  • Culture Media
  • Membrane Glycoproteins
  • Proteins
  • Oxidoreductases
  • TYRP1 protein, human
  • tyrosinase-related protein-1
  • Monophenol Monooxygenase
  • Isomerases
  • Intramolecular Oxidoreductases
  • dopachrome isomerase