Dynamics of process formation during differentiation of tectal neurons in embryonic zebrafish.

Neurons acquire their distinct shapes after passing through many transitional stages in early development. To reveal the dynamics and spatiotemporal sequence of process formation in situ, the growth of neurons in the optic tectum of live zebrafish embryos (54 to > 100 h old) was monitored using time-lapse videorecordings. Neurons were labeled by injecting the fluorescent vital dye DiO into the cell-rich layer of the developing tectum in 50- to 70-h-old embryos. In phase 1, tectal neurons possess an apical "primary process" which reaches to the ventral aspect of the tectal neuropil. The primary process produces at its tip short transitory branches, some with growth cones, over a period of roughly 6 h. One of the growth cones then elongates rapidly and grows toward the caudal tectum via a route characteristic of efferent axons. After retraction of excess branches and growth cones, branching activity resumes at the tip of the primary process to form the dendritic tree (phase 2). The dendritic tree develops in the tectal neuropil through emission and retraction of many branches during a period of > 20 h (our longest continuous time-lapse movie). The tectal territory "explored" in this way is larger than the area finally covered by the tree resulting from growth and loss of branches. The dynamics observed here directly are probably characteristic for dendrite formation in vertebrates. Moreover, consistent with the sequence of neuronal differentiation observed in vitro, the growth of the axon precedes that of the dendrites, although both emerge from a common primary process in this type of tectal neuron.