Morphometric analyses of changes in cell shape in the neuroepithelium of mammalian embryos

During neurulation the elevation of the neural folds is accompanied by, and thought to be partly driven by, changes in the shapes of the neuroepithelial cells. This paper explores the use of form factors for the measurement of changes in cell shape during normal and perturbed neural fold elevation in rat embryos; it is the first attempt to quantify changes in the shapes of cells during neurulation in mammalian embryos. Rat embryos at 9.9 days of gestation, when the neural epithelium is a biconvex plate, and at 10.4 days of gestation, when the cervical neural epithelium has formed the neural tube and when the cephalic neural folds have elevated but not fused, were used. In addition, embryos in which the cephalic neural folds had been caused to re-open by culture in saline with low calcium were examined. Embryos were fixed and then fractured across the neural axis. A montage of the fractured edge of the neuroepithelium was prepared from a series of scanning electron micrographs. Cells of the neuroepithelium were classified according to the shapes of their profiles as rectangular, round, apically tapered, basally tapered or spindle-shaped, the last three categories being considered as 'tapered'. The outlines of the cells were digitised using a computer-linked graphics tablet to provide estimations of the perimeters (p) and areas (a) of the cell profiles, and the form factors, f. of the cell profiles were calculated by f = 4 pi a/p2. This factor is a unitless ratio which is unity for a circle and less than unity for any other shape. It provides a measure of the efficiency with which a perimeter encloses an area, and thus indicates the degree of deformation of a shape from circular. From 9.9 to 10.4 days the cervical neuroepithelium shows a decrease in the percentage of rectangular cells and an increase in the percentage of spindle-shaped cells. The mean form factors of cells in the apically tapered and spindle-shaped categories decrease, showing that these cells are more elongated and tapered in later embryos. When embryos at 10.4 days are cultured in saline with low calcium the elevated cephalic neural folds collapse. Concomitantly the percentage of round cells in the cephalic neural folds increases and the percentage of apically tapered and spindle-shaped cells decrease.(ABSTRACT TRUNCATED AT 400 WORDS)