Shaping, invagination, and closure of the chick embryo otic vesicle: scanning electron microscopic and quantitative study.

Scanning electron microscopy, light microscopy, and morphometric analysis were used to study the morphological changes of the otic placode and vesicle before and during invagination and closure processes. Our results reveal that the otic placode undergoes shaping between stages HH9 and HH12; during this period the rostrocaudal axis is shortened, while the lateromedial axis of the placode lengthens. The presence of long cytokinesis bridges during this period suggests that cellular displacements after mitosis may participate in the shaping of the otic placode. The shaping process appears to facilitate the approach of the otic placode to the neural tube. From stage HH12 on, the otic anlage gradually becomes a U-shaped structure with its medial portion in intimate apposition to the rhombencephalic neural tube. The coincidence in time between the beginning of intimate otic anlage-rhombencephalon contact and active invagination suggests that these two processes are related. Changes occurring at the edges of the otic vesicle until their disappearance in stage HH17 suggest that, in addition to a process of invagination, the edges of the otic anlage become bent. During closure, cells at the edges of the otic vesicle differ in apical morphology according to their topographical location: The cells between the rostral and lateral edges have elongated apices, in contrast with the polygonal shape of the cell apices in other places of the edges. In the opposite side (between the caudal and medial edges) cell death is observed. Closure of the otic vesicle conceptualized as a zipper-like model is discussed. We propose that early development of the otic anlage takes place in four stages: 1) shaping (stages HH9-11); 2) triggering of the invagination (stage HH12); 3) early invagination and lateral bending (stages HH13-15); and 4) late invagination and closure (stages HH16-17).