Summary
Retinoids and growth factors seem to be important for normal mammalian reproduction and development. High levels of retinoic acid are teratogenic and induce cleft palate in the mouse. Little is known concerning the mechanisms through which retinoids induce cleft palate. Palatal epithelia from CD-1 embryonic mice on Day 12 of gestation were isolated from the mesenchyme and cultured in serum-free media, with all-trans retinoic acid or 13-cis retinoic acid, with or without epidermal growth factor (EGF). The epithelia attached and grew, and the cells differentiated over a 72-h culture period. Binding of [125I]EGF was observed in all cultures in a pattern that correlated with thymidine (TdR) uptake by the epithelia. EGF enhanced growth and [3H]TdR incorporation of the oral cells, but nasal cells generally did not proliferate. In this culture system, both retinoids suppressed [3H]TdR incorporation in a concentration-dependent manner for epithelia cultured with or without EGF. Medial cells are important to normal palatogenesis as they play a role in fusion of opposing shelves and subsequently many of these cells undergo programmed cell death. Death of medial cells in vitro is prevented by EGF and by the retinoids, either with or without EGF. This response occurs in the absence of a mesenchymal interaction, suggesting that the medial cell response to EGF and retinoids is not mediated by or dependent on the mesenchymal tissues. The survival of medial cells may be responsible for the failure of opposing shelves to fuse.
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Abbott, B. D.; Pratt, R. M. Retinoic acid and EGF induced alterationsin vitro andin vivo in the differentiation of the mouse palatal epithelium. Teratology 35: 31A-32A; 1987.
Abbott, B. D.; Adamson, E. D.; Pratt, R. M. Retinoic acid alters EGF receptor expression during palatogenesis. Development (in press, May), 1988.
Abbott, B. D.; Pratt, R. M. Retinoids and EGF alter embryonic mouse palatal epithelial and mesenchymal cell differentiation in organ culture. J. Craniofacial. Gen. Dev. Biol. 7: 219–240; 1987.
Adamson, E. D.; Deller, J. J.; Warshaw, J. B. Functional EGF receptors are present on mouse embryo tissues. Nature 291: 656–659; 1981.
Bakke, O. Antagonistic effect of glucocorticoids on retinoic acid induced growth inhibition and morphological alterations of a human cell line. Cancer Res. 46: 1275–1279; 1986.
Carpenter, G.; Cohen, S. Epidermal growth factor. Ann. Rev. Biochem. 48: 193–216; 1979.
Dion, L. D.; Gifford, G. E. Retinoic acid induces a G-1 cell cycle block in HeLa cells (49806). Proc. Soc. Exp. Biol. Med. 163: 510–514; 1980.
Fernhoff, P. M.; Lammer, E. J. Cranofacial features of Accutane embryopathy. J. Pediatr. 105: 595–597; 1984.
Geelen, J. A. G. Hypervitaminosis A induced teratogenesis. CRC Crit. Rev. Toxicol. 6: 351–375; 1979.
Greene, R. M.; Pratt, R. M. Inhibition by diazo-oxo-norleucine (DON) of rat palatal glycoprotein synthesis and epithelial cell adhesion in vitro. Exp. Cell Res. 105: 27–37; 1977.
Greene, R. M.; Pratt, R. M. Developmental aspects of secondary palate formation. J. Embryol. Exp. Morphol. 36: 225–245; 1976.
Grove, R. I.; Pratt, R. M. Influence of epidermal growth factor and cyclic-AMP on growth and differentiation of palatal epithelial cells in culture. Dev. Biol. 106:427–437; 1984.
Grove, R. I.; Pratt, R. M. Growth and differentiation of embryonic mouse palatal epithelial cells in primary culture. Exp. Cell Res. 148: 195–205; 1983.
Haddox, M. K.; Russell, D. H. Cell cycle-specific locus of vitamin A inhibition of growth. Cancer Res. 39: 2476–2480; 1979.
Haddox, M. K.; Scott, K. F.; Russell, D. H. Retinol inhibition of ornithine decarboxylase induction and G-1 progression in Chinese hamster ovary cells. Cancer Res. 39: 4930–4938; 1979.
Hall, J. G. Vitamin A: A newly recognized human teratogen. Harbinger of things to come? J. Pediatr. 105:583–584; 1984.
Hassell, J. R. The development of rat palatal shelves in vitro: an ultrastructural analysis of the inhibition of epithelial cell death and palate fusion by the epidermal growth factor. Dev. Biol. 45: 90–102; 1975.
Hassell, J. R.; Pratt, R. M. Elevated levels of cAMP alters the effect of epidermal growth factor in vitro on programmed cell death in the secondary palatal epithelium. Exp. Cell Res. 106: 55–62; 1977.
Jetten, A. M. Action of retinoids and phorbol esters on cell growth and the binding of epidermal growth factor. Ann. NY Acad. Sci. 359: 200–217; 1981.
Jetten, A. M. Retinoids specifically enhance the number of EGF receptors. Nature 284: 626–629; 1980.
Kochhar, D. M.; Johnson, E. M. Morphological and autoradiographic studies of cleft palate induced in rat embryos by maternal hypervitaminosis A. J. Embryol. Exp. Morphol. 14: 223–238; 1965.
Kochhar, D. M.; Penner, J. D.; Tellone, C. I. Comparative teratogenic activities of two retinoids: Effects on palate and limb development. Teratol. Carcinogen. Mutagen. 4: 377–387; 1984.
Lotan, R.; Nicolson, G. L. Inhibitory effects of retinoic acid or retinyl acetate on the growth of untransformed, transformed, and tumor cells in vitro. JNCI 59: 1717–1722; 1977.
Lotan, R.; Neumann, G.; Lotan, D. Characterization of retinoic acid-induced alterations in the proliferation and differentiation of a murine and a human melanoma cell line in culture. Ann. NY Acad. Sci. 359: 150–170; 1981.
LaBarca, C.; Paigen, K. A simple, rapid, and sensitive DNA assay procedure. Anal. Biochem. 102: 344–352; 1980.
Newall, D. R.; Edwards, J. R. G. The effect of vitamin A on fusion of mouse palates. I. Retinyl palmitate and retinoic acid in vivo. II. Retinyl palmitate, retinol, retinoic acid in vitro. Teratology 23: 115–130; 1981.
Nexo, E.; Hollenberg, M. D.; Figueroa, A., et al. Detection of epidermal growth factor-urogastrone and its receptor during fetal mouse development. Proc. Natl. Acad. Sci. USA 77(5):2782–2785; 1980.
Pratt, R. M.; Grove, R. I. Influence of epidermal growth factor on growth and differentiation of embryonic mouse palatal epithelial cells in vitro. Cell Differ. 16: 52S; 1985.
Pratt, R. M.; Yoneda, T.; Silver, M. H., et al. Involvement of glucocorticoids and epidermal growth factor in secondary palate development. In: Pratt, R. M.; Christiansen, R. L., eds. Current research trends in prenatal craniofacial development. New York: Elsevier/North-Holland; 1980:235–252.
Rees, A. R.; Adamson, E. D.; Graham, C. F. Epidermal growth factor receptors increase during the differentiation of embryonal carcinoma cells. Nature 281: 309–311; 1979.
Roberts, A. B.; Sporn, M. B. Cellular biology and biochemistry of the retinoids, vol. 2. In: Sporn, M. B.; Roberts, A. B.; Goodman, D. S., eds. New York: Academic Press; 1984: 209–286.
Snedecor, G. W.; Cochran, W. G. Statistical methods. Ames, Iowa: Iowa State University Press; 1967.
Tyler, M. S.; Pratt, R. M. Effect of epidermal growth factor on secondary palatal epithelium in vitro: Tissue isolation and recombination studies. J. Embryol. Exp. Morphol. 58: 93–106; 1980.
Yoneda, T.; Pratt, R. M. Mesenchymal cells from the human embryonic palate are highly responsive to epidermal growth factor. Science 213: 563–565; 1981.
Yuspa, S. H.; Elgjo, K.; Morse, M. A., et al. Retinyl acetate modulation of cell growth kinetics and carcinogen-cellular interaction in mouse epidermal cell cultures. Chem. Biol. Interac. 16: 251–264; 1977.
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Abbott, B.D., Pratt, R.M. Influence of retinoids and EGF on growth of embryonic mouse palatal epithelia in culture. In Vitro Cell Dev Biol 24, 343–352 (1988). https://doi.org/10.1007/BF02628837
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DOI: https://doi.org/10.1007/BF02628837