Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Rapid changes in the extracellular matrix accompany in vitro palatal shelf remodelling

  • 28 Accesses

  • 8 Citations


The sequence of events and the distribution of extracellular matrix (ECM) components was examined during mouse secondary palatal shelf elevation in an in vitro model using standard roller tube culture methods developed for the culture of early embryos. In this culture system, the morphological changes associated with remodelling and reorientation of the palatal shelves of gestational day 13 mouse fetuses were similar to those observed in vivo. However, in specimens explanted 24–30 h prior to reorientation in vivo, remodelling began rapidly after explantation, and significant reorientation was accomplished within 4 h. Midline contact between the shelves did not occur until after 18 h in vitro, concomitant with shelf growth. Therefore, in this in vitro model, events related to palatal shelf remodelling and reorientation can be distinguished from those related to shelf growth. We used this in vitro model to characterize the transient changes in ECM distribution and accumulation that occur concomitant with events in shelf remodelling. Our results show that, during rapid remodelling in vitro, the relative distributions of collagen III, fibronectin and hyaluronate, as visualized by immunofluorescent staining, decreased within specific regions of the mesenchymal compartment. In contrast, the distribution of collagen I within the mesenchyme increased, and the distribution of tenascin did not change significantly. All molecules examined, except tenascin, showed changes in distribution within the basement membrane. These patterns of distribution are similar to those observed during more gradual remodelling in vivo. During remodelling in vitro, the deposition of [3H]-glucosamine- and [3H]-proline-labelled components of the ECM, as visualized by autoradiography, was greatest during the first 3 h of culture. During this period, labelled ECM accumulated within specific regions of the mesenchyme and palatal epithelial basement membrane. Uptake was reduced dramatically during the subsequent 3 h in culture and was restricted mainly to the palatal epithelium and its underlying basement membrane. The in vitro system permitted the characterization of early events in shelf remodelling leading to reorientation. Results suggest that remodelling is accompanied by rapid, local accumulation of ECM in specific regions of the palatal shelf previously thought to play a role in the process.

This is a preview of subscription content, log in to check access.


  1. Brinkley L (1980) In vitro studies of palatal shelf elevation. In: Pratt R, Christiansen R (eds) Current research trends in prenatal craniofacial development. Elsevier North Holland, New York, pp 204–220

  2. Brinkley L (1984) Changes in cell distribution during mouse secondary palate closure in vivo and in vitro. I. Epithelial cells, Dev Biol 102:216–227

  3. Brinkley L, Morris-Wiman J (1987) Computer-assisted analysis of hyaluronate distribution during morphogenesis of the mouse secondary palate. Development 100:629–635

  4. Brinkley L, Vickerman M (1982) The effects of chlorcyclizine-induced alterations of glycosaminoglycans on mouse palatal shelf elevation in vivo and in vitro. J Embryol Exp Morphol 69:193–213

  5. Brinkley L, Basehoar B, Branch A, Avery J (1975) A new in vitro system for studying secondary palatal development. J Embryol Exp Morhol 34:485–495

  6. Diewert V (1974) A cephalometric study of orofacial structures during secondary palate closure in the rat. Arch Oral Biol 19:303–315

  7. Diewert V, Tait B (1979) Palatal process movement in the rat as demonstrated in frozen sections. J Anat 128:609–618

  8. Ferguson M (1988) Palate development. Development 103 [Suppl]:41–69

  9. Foreman D, Sharpe P, Ferguson M (1991) Comparative biochemistry of mouse and chick secondary palate development in vivo and in vitro with particular emphasis on extracellular matrix molecules and the effects of growth factors on their synthesis. Arch Oral Biol 36:457–471

  10. Green S, Tarone G, Underbill C (1988) Distribution of HA and HA-receptors in the adult lung. J Cell Sci 89:145–156

  11. Knudsen T, Bulleit R, Zimmerman E (1988) Histochemical localization of glycosaminoglycans during morphogenesis of the secondary palate in mice. Anat Embryol 173:137–142

  12. Eewis C, Thibault L, Pratt R, Brinkley E (1980) An improved culture system for secondary palate elevation. In Vitro 16:453–460

  13. Morris-Wiman J, Brinkley L (1992) An extracellular matrix infrastructure provides support for murine secondary palatal shelf remodelling. Anat Rec 234:575–586

  14. New D, Coppola P, Terry S (1973) Culture of explanted rat embryos in rotating tubes. J Reprod Fertil 35:1135–1138

  15. Pratt R, King C (1971) Collagen synthesis in the secondary palate of the developing rat. Arch Oral Biol 16:1181–1185

  16. Pratt R, Goggins J, Wilk A, King C (1973) Acid mucopolysaccharide synthesis in the secondary palate of the developing rat at the time of rotation and fusion. Dev Biol 32:230–237

  17. Sharpe P, Ferguson M (1988) Mesenchymal influences of epithelial differentiation in developing systems. J Cell Sci [Suppl-10]:195–230

  18. Silver M, Foidart J-M, Pratt R (1981) Distribution of fibronectin and collagen during mouse limb and palate development. Differentiation 18:141–149

  19. Smiley G, Koch W (1972) An in vitro and in vivo study of single palatal processes. Anat Rec 73:405–416

  20. Toole B (1981) Glycosaminoglycans in morphogenesis. In: Hay E (ed) Cell biology of the extracellular matrix. Plenum Press. New York, pp 259–294

  21. Walker B (1969) Correlation of embryonic movement with palate closure in mice. Teratology 2:191–198

  22. Walker B, Crain F (1960) Effects of hypervitaminosis A on palate development in two strains of mice. Am J Anat 107:49–58

  23. Walker B, Fraser C (1956) Closure of the secondary palate in three strains of mice. J Embryol Exp Morphol 4:176–189

  24. Walker B, Quarles J (1976) Palate development in mouse foetuses after tongue removal. Arch Oral Biol 21:405–412

  25. Wilk A, King C, Pratt R (1978) Chlorcyclizine induction of cleft palate in the rat: degradation of palatal glycosaminoglycans. Teratology 28:199–210

Download references

Author information

Correspondence to J. Morris-Wiman.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Morris-Wiman, J., Brinkley, L. Rapid changes in the extracellular matrix accompany in vitro palatal shelf remodelling. Anat Embryol 188, 75–85 (1993). https://doi.org/10.1007/BF00191453

Download citation

Key words

  • Mouse
  • Palate
  • Extracellular matrix
  • In vitro