Ultrastructural changes of statoconia after segmentation of the otolithic membrane

The chick vestibule transformed from a homogeneous epithelial layer at day 2 (stage 15) into a pseudo-stratified epithelial layer at day 4 (stage 24). The apical columnal appearance of sensory cells was evident by day 6 (stage 29). In the supporting cells of the saccule and utricle large rough endoplasmic reticulum cisterns filled with material similar to the primitive organic matrix. Fibrillar material of the otolithic membrane remained attached to the supporting cells and accumulated over the saccule and utricle. The primitive otolithic membrane acquired stress-like lines and statoconial units emerged from the upper surface without a central core. Statoconia thickened at the periphery and a central core formed. Calcium was deposited between the fibrils of older statoconia which were located on top of the segmenting membrane. DIAMOX inhibited statoconia formation and/or prevented calcium and the matrix from associating. Large statoconia (100-200 microns diameter) were formed in embryos injected with this carbonic anhydrase inhibitor. Gel electrophoresis of immature statoconial complexes yielded at least 5 major protein bands between 25 and 210 kDa. Ouabain-sensitive potassium-dependent p-nitrophenylphosphatase activity was demonstrated in the endolymphatic sac of newly hatched chicks.