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Short term retinol treatment in vitro induces stable transdifferentiation of chick epidermal cells into mucus-secreting cells

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Epidermal mucous metaplasia of cultured skin can be induced by treatment with excess retinol for several days (Fell 1957). In the induction of mucous metaplasia, retinol primarily affects the dermal cells and retinol-pretreated dermis can alter epidermal differentiation towards secretory epithelium (Obinata et al. 1987). In this work, we found that mucous metaplasia could be induced by culturing 13-day-old chick embryonic tarsometatarsal skin in medium containing retinol (20 μM) for only 8–24 h, followed by culture in a chemically defined medium (BGJb) without retinol or serum for 6 days. The application of cycloheximide together with retinol during the first 8 h of culture inhibited epidermal mucous metaplasia during subsequent culture for 6 days in BGJb, indicating that induction of a signal(s) in the dermis by excess retinol requires protein synthesis. However, the presence of 20 nM hydrocortisone (Takata et al. 1981) throughout the culture period did not inhibit retinol-induced epidermal mucous metaplasia of the epidermis. This indicates that a brief treatment of the skin with excess retinol determines the direction of epithelial differentiation toward secretory epithelium; this is a simpler in vitro system for the induction of epidermal mucous metaplasia than those established before.

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  1. Antonopoulos CA, Gardell JA, Szirmai JA, Tyssonsk ER (1964) Determination of glycosaminoglycans (mucopolysaccharides) from tissues on the microgram scale. Biochim Biophys Acta 83:1–19

  2. Ashwell G (1963) Determination of pentoses. In: Colowick SP, Kaplan NO (eds) Methods in enzymology. (Vol 3) Academic Press, New York, pp 87–90

  3. Asselineau D, Bernard BA, Bailly C, Darmon MD (1989) Retinoic acid improves epidermal morphogenesis. Dev Biol 133:322–335

  4. Barnett ML, Szabo G (1973) Effect of vitamin A on epithelial morphogenesis in vitro. Fine structural changes in explant of adult mammalian skin. Exp Cell Res 76:118–126

  5. Biggers JD, Gwatkin RBL, Heyner S (1961) Growth of embryonic avian and mammalian tibiae on a relatively simple chemically defined medium. Exp Cell Res 25:41–58

  6. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

  7. Burton K (1956) A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J 62:315–323

  8. Fell HB (1957) The effect of excess vitamin A on culture of embryonic chicken skin explanted at different stages of differentiation. Proc R Soc London B 146:242–256

  9. Fuchs E, Green H (1981) Regulation of terminal differentiation of cultured human keratinocytes by vitamin A. Cell 25:617–625

  10. Hirano H, Oishi N, Obinata A, Endo H (1985) Metaplastic changes in chick embryonic skin induced by vitamin A. In: Shibata S (ed) Basement membranes. Elsevier, Amsterdam New York Oxford, pp 395–406

  11. Kalliecharan R, Hall BK (1974) A developmental study of the levels of progesterone, corticosterone, cortisol and cortisone circulating in plasma of chick embryos. Gen Comp Endocrinol 24:364–372

  12. Lichti U, Ben T, Yuspa SH (1985) Retinoic acid-induced transglutaminase in mouse epidermal cells is distinct from epidermal transglutaminase. J Biol Chem 260:1422–1426

  13. Linsley PS, Kallestad JC, Horn D (1988) Biosynthesis of high molecular weight breast carcinoma associated mucin glycoproteins. J Biol Chem 263:8390–8397

  14. Obinata A, Kawada M, Endo H (1987) Induction of epidermal mucous metaplasia by culture of recombinants of undifferentiated epidermis and retinol-treated dermis in a chemically defined medium. Dev Biol 123:59–62

  15. Obinata A, Akimoto Y, Hirano H, Endo H (1991) Stimulation by Bt2cAMP of epidermal mucous metaplasia in retinol-pretreated chick embryonic cultured skin, and its inhibition by herbimycin A, an inhibitor for proteintyrosine kinase. Exp Cell Res 193:36–44

  16. Oda Y, Ohyama Y, Obinata A, Endo H, Kasai K (1989) Endogenous galactoside-binding lectin expression is suppressed in retinol-induced mucous metaplasia of chick embryonic epidermis. Exp Cell Res 182:33–43

  17. Pamer T, Glass GBJ, Horowitz MI (1968) Purification and characterization of sulfated glycoproteins and hyaluronidase-resistant mucopolysaccharides from dog gastric mucosa. Biochem 7:3821–3829

  18. Peck GL, Elias PM, Wetzel B (1977) Effects of retinoic acid on embryonic chick skin. J Invest Dermatol 69:463–476

  19. Pelc SR, Fell HB (1960) The effect of excess vitamin A on the uptake of labelled compounds by embryonic skin in organ culture. Exp Cell Res 19:99–113

  20. Petkovich M, Brand NJ, Kurst A, Chambon P (1987) A human retinoic acid receptor which belongs to the family of nuclear receptors. Nature 330:444–450

  21. Rask L, Peterson PA (1976) In vitro uptake of vitamin A from the retinol-binding plasma protein to mucosal epithelial cells from the monkey's small intestine. J Biol Chem 251:6360–6366

  22. Rothblat GH, Arborgast LY, Quellett L, Howard BV (1976) Preparation of delipidized serum protein for use in cell culture system. In Vitro 12:554–557

  23. Saarni H, Tammi M (1977) A rapid method for separation and assay of radiolabeled mucopolysaccharides from cell culture medium. Anal Biochem 81:40–46

  24. Schmidt G, Thannhauser J (1945) A method for the determination of desoxyribonucleic acid, ribonucleic acid, and phosphoproteins in animal tissues. J Biol Chem 161:83–89

  25. Shapiro SS, Poon JP (1978) Effect of retinyl acetate on sulfated glycosaminoglycan biosynthesis in dermal and epidermal cells in vitro. Connective Tissue Res 6:101–108

  26. Takata K, Obinata A, Endo H, Hirano H (1981) Induction of the alpha-type keratinization by hydrocortisone in embryonic chick skins grown in a chemically defined medium. An electron microscopic study. Dev Biol 85:370–379

  27. Thacher SM, Coe EL, Rice RM (1985) Retinoid supression of transglutaminase activity and envelope competence in cultured human epidermal carcinoma cells. Differentiation 29:82–87

  28. Tseng SCG, Hatchell D, Tierney N, Huang AJW, Sun TT (1984) Expression of specific keratin markers by rabbit corneal, conjunctival, and esophageal epithelia during vitamin A deficiency. J Cell Biol 99:2279–2286

  29. Wilkoff LJ, Peckham JC, Dulmadge EA, Mowry RW, Chopra DP (1976) Evaluation of vitamin A analogs in modulating epithelial differentiation of 13-day chick embryo metatarsal skin explants. Cancer Res 36:964–972

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Obinata, A., Akimoto, Y., Hirano, H. et al. Short term retinol treatment in vitro induces stable transdifferentiation of chick epidermal cells into mucus-secreting cells. Roux's Arch Dev Biol 200, 289–295 (1991). https://doi.org/10.1007/BF00241298

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Key words

  • Retinol
  • Hydrocortisone
  • Mucous metaplasia
  • Transdifferentiation
  • Epidermis