Rapid disappearance of the medial epithelial seam during palatal fusion occurs by multifocal breakdown that is preceded by expression of alpha smooth muscle actin in the epithelium.
Breakdown of the medial epithelial seam (MES) is essential to allow bridging of the mesenchyme during palatal fusion. Evidence exists for three mechanisms for this breakdown that are incompatible at the level of individual cells in the seam. To determine if breakdown of the seam was regionally restricted, 3-dimensional reconstructions were generated using volume rendering software from 1 micron serial sections in the sagittal plane of rat palates fixed during the process of fusion. The earliest break detected in electron micrographs was cell separation and in reconstructions was a discrete defect, with a rounded outline, nearer to the nasal than to the oral margin of the seam. Further breakdown produced a pattern of rounded defects along the nasal margin of the seam resulting in interconnected columns of cells preferentially attached to the oral epithelium. Computer generated slicing of reconstructed seams showed that groups of cells evident in cross-sections as islands at this stage of breakdown of the MES could be artifacts. Unequivocal islands of epithelial cells formed later in fusion had a rounded outline, an incomplete basal lamina and a halo of cells containing phagocytosed apoptotic debris. The pattern of breakdown indicated that the MES breaks down under tension. Laser confocal microscopy of sections and whole-mounts of palates demonstrated alpha-smooth muscle actin preferentially localized in the epithelial cells of the palatal shelves immediately before and during formation of the seam. Expression in epithelial cells of the isoform of actin normally restricted to smooth muscle cells engaged in tonic contraction supported an interpretation that the epithelial cells of the seam may be capable of generating tension during the palatal fusion event.
Gibbins
J R
JR
Department of Pathology, The University of Sydney, Australia. johngi@pathology.usyd.edu.au
Brent
S
S
Srivastava
M
M
Garibotto
N
N
Tazawa
Y M
YM
Cameron
A
A
Hunter
N
N
D000199:Actins / Q000096:biosynthesis*; D000818:Animals; D001485:Basement Membrane / Q000196:embryology; D004848:Epithelium / Q000196:embryology / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D017403:In Situ Hybridization; D008854:Microscopy, Electron; D010159:Palate / Q000196:embryology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D000199:Actins