Surface coat material associated with the developing otic placode/vesicle in the chick.

Surface coat material (SCM) has been illustrated in association with the apical surfaces of numerous epithelia during morphogenesis. This study investigates the development of a SCM associated with the invaginating otic placode/vesicle in the chick. Glycoconjugate containing SCM was retained by the inclusion of cetylpyridinium chloride (CPC) in the fixative, histochemically visualized by using ruthenium red (RR) staining, and viewed by scanning (SEM) or transmission (TEM) electron microscopy. Initial characterization of the glycoconjugates present in this material was elucidated by using lectins conjugated to fluorescein isothiocyanate. Lectins utilized included concanavalin A (Con A), wheat germ agglutinin (WGA), and soybean agglutinin (SBA). Invagination of the otic placode was apparent as early as stage 12. By stage 15 the vesicle was beginning to separate from the surface ectoderm as evidenced by its aperture, which was altered in shape and reduced in size. All embryos fixed with glutaraldehyde containing either CPC or RR were shown to possess SCM associated with the surface ectoderm, particularly in the area of the otic placode/vesicle. Additional embryos were processed by cryofixation without prior aldehyde fixation; these also exhibited SCM. All lectins labelled the epithelium of the otic placode/vesicle. However, their binding patterns were not identical. The binding of Con A and WGA remained constant over the stages studied, while SBA increased as the otic vesicle developed. The data clearly indicate that otic placode morphogenesis is accompanied by the synthesis of SCM rich in glycoconjugates.