Transcription of melanogenesis enzymes in melanocytes: dependence upon culture conditions and co-cultivation with keratinocytes.
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.
Kippenberger
S
S
Abteilung 1, Klinikum der Johann Wolfgang Goethe-Universität, Frankfurt/Main, Germany.
Bernd
A
A
Bereiter-Hahn
J
J
Ramirez-Bosca
A
A
Kaufmann
R
R
Holzmann
H
H
D018920:Coculture Techniques; D003470:Culture Media; D006801:Humans; D019746:Intramolecular Oxidoreductases; D007535:Isomerases / Q000235:genetics*; D015603:Keratinocytes; D008544:Melanocytes / Q000201:enzymology*; D008562:Membrane Glycoproteins; D014442:Monophenol Monooxygenase / Q000235:genetics*; D010088:Oxidoreductases; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000235:genetics*; D012867:Skin / Q000196:embryology / Q000201:enzymology / Q000473:pathology; D014158:Transcription, Genetic
D003470:Culture Media; D008562:Membrane Glycoproteins; D011506:Proteins; D010088:Oxidoreductases; C489799:TYRP1 protein, human; C064056:tyrosinase-related protein-1; D014442:Monophenol Monooxygenase; D007535:Isomerases; D019746:Intramolecular Oxidoreductases; C042501:dopachrome isomerase