First steps of otolith formation of the zebrafish: role of glycogen?

The first steps of otolith formation were studied by electron microscopy in zebrafish embryos at different postfertilization (PF) time intervals. Between 19 and 22 h PF, the otic cavity contains glycogen particles derived by an apocrine process from the apical portions of the epithelial cells of the inner ear. The particles are arranged in parallel arrays, then in pseudocrystalloid structures, and finally in concentric arrays to form dense clusters referred to as "spherules". At 23 h PF, a group of "globules", consisting of modified aggregated "spherules" surrounded by several free "spherules", forms the nascent otolith. At 30 h PF, fused globules form a roughly spherical otolith. Spherules undergoing their process of modification and aggregation, are located in its central part, and constitute the so-called "nucleus". At 50 h PF, the otolith is a flattened hemisphere. It is made up of fused globules surrounded by two concentric layers whose organization is similar to that observed in the otolith of the adult fish. At this stage, calcium may be detected in the otolith except in its nucleus. We suggest that glycogen molecules found in the nascent otolith might allow the insertion of molecules such as glycoproteins (collagens) which are known to fix calcium. As a result, glycogen might play a key role in initiating the formation of otoliths and possibly that of other calcified tissues.

Chronology of the appearance of beta, A, and alpha mitochondria-rich cells in the gill epithelium during ontogenesis of the brown trout (Salmo trutta).

Three types of mitochondria-rich (MR) cells, the alpha, beta, and accessory cells, are observed in the gill epithelium of juvenile and adult freshwater teleosts. In addition to numerous mitochondria, their cytoplasm contains a network of membranous tubules, the tubular system, connected to the laterobasal plasma membrane. Because they are believed to play a role in ionic regulation, it is of interest to examine the order of appearance and the ultrastructural characteristics of such cells during the embryogenesis and larval life of the brown trout. Gills of embryos and fry maintained in freshwater were thus removed at different stages and prepared for transmission and scanning electron microscopic examination. One week before hatching, cells resembling the beta cells of juvenile and adult teleosts appeared first among the epithelial cells located at the base of the filaments in the gills of the brown trout larva. In addition to their tubular system, they contained numerous and large apical structures seemingly originating from the Golgi apparatus. At approximately hatching time, small pear-shaped cells were seen to be closely apposed to the lateral side of the beta cells; they were usually devoid of apical structures and were considered to be accessory cells. After yolk sac resorption, additional cells, the alpha cells, were present along the lamellae. In contrast to the beta cells, they only exhibited poorly developed apical structures. The possible role of these three types of MR cells in osmoregulation during fish development is discussed.