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Fetuses were obtained from CD-1 mice at a time estimated to be 12 h prior to vivo secondary palate closure. One of the palatal shelves of each partially dissected fetal head was lesioned in one of five ways, the other left intact to serve as control. Single transverse cuts extending the width of the shelf were made at one of three positions along the longitudinal axis of the shelf: one-third, one-half or two-thirds the shelf length estimated from the rostral edge. Some specimens were cut in two places, dividing the shelf into three equal segments. Another group received a lesion which separated the caudal third of the shelf from its maxillary connections. All specimens were cultured for 18 h. At the end of the culture period the heads were fixed, examined and the degree of elevation of each shelf piece assessed. Intact, control shelves of all preparations were elevated in the rostral two-thirds of the shelf, while the caudal third was partially elevated. Results seen in lesioned shelves depended upon both the size of the segment and the region of the shelf contained in the segment. The rostral two-thirds of the shelf, the presumptive hard palate, whether intact or in segments elevated without physical connections to neighboring shelf tissue. Thus, it is unlikely that this elevation requires a wave of contraction be transmitted from the caudal soft palate region. In contrast, the presumptive soft palate requires continuity with the rostral portions of the shelf both to maintain structural stability and to elevate.

The intrinsic forces necessary for directing the reorientation of the secondary palate appear to reside in the anterior two thirds of the palate or presumptive hard palate. The hard palate could reorient regardless of whether it was intact or separated from the posterior third or presumptive soft palate. The soft palate could only reorient if the palate shelves are left intact. These intrinsic forces, within the hard palate, may be mediated by the mesenchymal cells, their extracellular matrix, or the epithelium surrounding the shelves. This latter possibly was tested by removing the epithelium, from either the presumptive oral or nasal surface followed by measurement of reorientation in vitro. Only after removal of the oral epithelium was a significant inhibition in reorientation observed. The treatment used to remove the epithelium, EDTA and scraping, was shown to remove 41% of the oral epithelium leaving the majority of the basement membrane intact. The observed inhibition of reorientation did not appear to be a consequence of wound healing. Creation of wounds twice the area that was observed after treatment with EDTA and scraping inhibited reorientation minimally. These results suggest that the epithelium and particularly the anterior oral epithelium plays a major role in the reorientation of the murine secondary palate.

The patterns of distribution of both total mesenchymal cells and the ratios of [3H]thymidine-labelled to total cells were mapped during secondary palatal shelf reorientation in vivo and in vitro. Smoothed spatial averaging, a computer-assisted method which takes into account the positions of all cells across an entire histological section of the shelf, was employed. Changes in shelf cross-sectional area and cell size were also measured. Three shelf regions, anterior and posterior presumptive hard and presumptive soft palate, were studied at developmental stages which were 30, 24 and 18 h prior to expected in vivo elevation, after in vivo reorientation and during the course of in vitro reorientation. Region-specific patterns of cell distribution change with shelf reorientation. These changes were observable within 6 h. Increases in cell number by cell division may enhance some high local cell densities, but cannot account for decreases in cell density. Increase in cell size is not a factor in decreasing cell density, nor is cell death. Displacement of cells by expansion of the extracellular matrix may be involved.