Summary
Primary cultures of guinea pig ear keratinocytes were treated with vitamin A acid at plating, or at 14 h or 14 days after plating. The intracellular content of the cyclic nucleotides cAMP and cGMP was determined by radioimmunoassay at intervals during a period of 50 h after treatment. When added at plating, vitamin A acid produced a wave of DNA synthesis and increase in DNA which was at maximum between 30 and 40 h after plating, and coincided with decreased cAMP levels. This may represent a subpopulation of keratinocytes in S phase. Treatment with vitamin A acid at 14 h or 14 days after plating resulted in an immediate but temporary fall in cAMP and cGMP, and a wave of thymidine uptake but no increase in DNA per dish. Thus, a single treatment with vitamin A acid is mitogenic only when applied at plating. At other times, it can cause changes in cyclic nucleotide content without any observable cell proliferation.
Zusammenfassung
Primärkulturen von Meerschweinchenohr-Keratinocyten wurden mit Vitamin A-Säure behandelt zum Zeitpunkt des Ansetzens der Kulturen oder 14 h bzw. 14 Tage danach. In einem Zeitraum bis 50 h nach Behandlung wurde die intracelluläre Konzentration von den cyclischen Nukleotiden cAMP und cGMP mit einem Radio-Immunoassay in Intervallen bestimmt. Wenn die Zugabe der Vitamin A-Säure beim Ansetzen der Kulturen erfolgte, resultierte eine Welle von DNA-Synthese und eine Zunahme der DNA-Konzentration, die ihre Maxima zwischen 30 und 40 h nach Kulturansatz erreichte und zeitlich mit einer Abnahme der cAMP-Konzentration zusammenfällt. Dies könnte auf eine Keratinocyten-Subpopulation hinweisen. Die Zugabe von Vitamin A-Säure 14 h oder 14 Tage nach Kulturansatz führte zu sofortiger, wenn auch zeitlich begrenzter Abnahme der cAMP- und cGMP-Konzentration sowie einer Welle von Thymidin-Aufnahme die nicht mit einer Zunahme des DNA-Gehalts in der Petrischale verbunden war. Somit wirkt eine einmalige Behandlung mit Vitamin A-Säure nur dann Mitoseauslösend, wenn deren zugabe beim Ansetzen der Zellkultur erfolgte. Zu späteren Zeitpunkten verursacht Vitamin A-Säure Veränderungen in der Konzentration der cyclischen Nukleotide ohne bemerkbare Zellproliferation.
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Supported by United States Public Health Service Grant AM 15107
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Wilkinson, D.I., Orenberg, E.K. Effect of vitamin A acid on cyclic nucleotides of cultured keratinocytes. Arch Dermatol Res 267, 25–31 (1980). https://doi.org/10.1007/BF00416918
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DOI: https://doi.org/10.1007/BF00416918