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Localization of vitamin a by autofluorescence during induced metaplastic changes in cultures of skin

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Summary

A technique was devised for following the uptake and location of vitamin A in organ cultures. Explants of 12- and 13-day embryonic mouse upper lip skin were grown for 3,6 or 9 days in biological medium to which was added 0,4.1 or 6.9 μg per ml of retinyl acetate. This form of vitamin A caused glandular morphogenesis of vibrissa follicles, and keratinization in epidermis and follicles was completely suppressed in 12-day explants and partially suppressed in 13-day explants. Frozen sections at 16 μm showed the white, non-fading fluorescence of keratin and the green, rapidly-fading fluorescence due to vitamin A which was captured by high-speed photography. Although more concentrated within lipid droplets in the dermis, the vitamin penetrated both the epidermis and the hair follicles. The ability to obtain permanent photographic records of the fading fluorescence makes this a useful method for analyzing vitamin A distribution as well as keratin distribution.

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References

  1. Fell, H. B., and E. Mellanby. 1953. Metaplasia produced in cultures of chick ectoderm by high vitamin A. J. Physiol. (Lond.) 119: 470–488.

    CAS  Google Scholar 

  2. New, D. A. T. 1963. Effects of excess vitamin A on cultures of skin and buccal epithelium of the embryonic rat and mouse. Br. J. Dermatol. 75: 320–325.

    Article  PubMed  CAS  Google Scholar 

  3. Hardy, M. H. 1967. Responses of embryonic mouse skin to excess vitamin A in organotypic cultures from the trunk, upper lip and lower jaw. Exp. Cell Res. 46: 367–384.

    Article  CAS  Google Scholar 

  4. Hardy, M. H. 1968. Glandular metaplasia of hair follicles and other responses to vitamin A excess in cultures of rodent skin. J. Embryol. Exp. Morphol. 19: 157–180.

    PubMed  CAS  Google Scholar 

  5. Bellows, C. G., and M. H. Hardy. 1977. Histochemical evidence of mucosubstances in the metaplasic epidermis and hair follicles produced in vitro in the presence of excess vitamin A. Anat. Rec. 187: 257–272.

    Article  PubMed  CAS  Google Scholar 

  6. Popper, H. 1941. Histologic distribution of vitamin A in human organs under normal and pathologic conditions. Arch. Pathol. Lab. Med. 31: 766–802.

    CAS  Google Scholar 

  7. Grüneberg, H. 1943. The development of some external features in mouse embryos. J. Hered. 34: 88–92.

    Google Scholar 

  8. Hardy, M. H. 1978. The Maximow single cover slip technique for organ culture. Tissue Culture Association Manual 4: 889–892.

    Article  Google Scholar 

  9. Guyer, M. H. 1953.Animal Micrology, University of Chicago Press, Chicago, Illinois, pp. 239–240.

    Google Scholar 

  10. Carter, H. B., and W. H. Clarke. 1957. The hair follicle group and skin follicle population of Australian merino sheep. Aust. J. Agric. Res. 8: 91–108.

    Article  Google Scholar 

  11. Hardy, M. H. 1969. The differentiation of hair follicles and hairs in organ culture. In: W. Montagna and R. L. Dobson (Eds.),Advances in Biology of Skin. Vol. 9:Hair Growth. Pergamon press, Oxford, pp. 35–60.

    Google Scholar 

  12. Davidson, P., and M. H. Hardy. 1952. The development of mouse vibrissaein vivo andin vitro. J. Anat. 86: 342–356.

    PubMed  CAS  Google Scholar 

  13. Sweeny, P. R., and M. H. Hardy. 1976. Ciliated and secretory epidermis produced from embryonic mammalian skin in organ culture by vitamin A. Anat. Rec. 185: 93–100.

    Article  PubMed  CAS  Google Scholar 

  14. Udenfriend, S. 1962.Fluorescence Assay in Biology and Medicine. Academic Press, New York, London, p. 517.

    Google Scholar 

  15. Cornbleet, T., and H. Popper. 1942. Properties of human skin revealed by fluorescence microscopy. Arch. Dermatol. Syphilol. 46: 59–65.

    CAS  Google Scholar 

  16. Yamamoto, M., H. Enzan, H. Hara, and S. Iijima. 1978. Fluorescence and microscopic studies on the perivascular mesenchymal cells and fibroblasts after vitamin A administration. Acta Pathol. Japn. 28: 513–521.

    CAS  Google Scholar 

  17. Popper, H., and R. Greenberg. 1941. Visualization of vitamin A in rat organs by fluorescence microscopy. Arch. Pathol. Lab. Med. 32: 11–32.

    CAS  Google Scholar 

  18. Hardy, M. H., P. R. Sweeny, and C. G. Bellows. 1978. The effects of vitamin A on the epidermis of the fetal mouse in organ culture: An ultrastructural study. J. Ultrastruct. Res. 64: 246–260.

    Article  PubMed  CAS  Google Scholar 

  19. Kutsky, R. J. 1973.Handbook of Vitamins and Hormones. Van Nostrand Reinhold Co., New York, pp. 7–15.

    Google Scholar 

  20. Sherman, B. S. 1967. Uptake and autoradiographic demonstration of vitamin A and/or its metabolic derivatives in organ cultures of rat prostate. Experientia 23: 353–355.

    Article  PubMed  CAS  Google Scholar 

  21. Rask, L., and P. A. Peterson. 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.

    PubMed  CAS  Google Scholar 

  22. Ong, D. E., and F. Chytil. 1975. Specificity of cellular retinol-binding protein from compounds with vitamin A activity. Nature 255: 74–75.

    Article  PubMed  CAS  Google Scholar 

  23. Sani, B. P., and D. L. Hill. 1976. A retinoic acid-binding protein from chick embryo skin. Cancer Res. 36: 409–413.

    PubMed  CAS  Google Scholar 

  24. Plopper, C. G., D. L. Wallace, and J. D. Pietsch. 1975. Extraction of C-14-vitamin A from rat liver and kidney during processing for transmission electron microscopy. Stain Technol. 50: 339–345.

    PubMed  CAS  Google Scholar 

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This work was supported by the National Research Council of Canada (Operating Grant to M. H. Hardy and Postgraduate Scholarship to R. Van Exan) and by the Ontario Ministry of Agriculture and Food.

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Vanexan, R.J., hardy, M.H. Localization of vitamin a by autofluorescence during induced metaplastic changes in cultures of skin. In Vitro 15, 631–640 (1979). https://doi.org/10.1007/BF02618264

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  • DOI: https://doi.org/10.1007/BF02618264

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