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Journal of Neuroscience, Vol 12, 467-482, Copyright © 1992 by Society for Neuroscience
Axonogenesis and morphogenesis in the embryonic zebrafish brain
LS Ross, T Parrett and SS Easter Jr
Biology Department, University of Michigan, Ann Arbor 48109-1048.
We have examined early neuronal differentiation and axonogenesis in the
fore- and midbrain of zebrafish embryos to address general issues of early
vertebrate brain development. AChE expression and HNK-1 antibody
immunoreactivity were used as markers for differentiated neurons and axons,
respectively. The pattern of neuronal differentiation followed a
stereotyped sequence. AChE-positive cells first appeared between 14 and 16
hr in three small, isolated, bilaterally symmetrical clusters on the
surface of the brain. The three clusters--the dorsorostral, ventrorostral,
and ventrocaudal clusters--proved to be the progenitors of the
telencephalon, ventral diencephalon, and mesencephalic tegmentum,
respectively. With further development, more cells were added to these
three clusters, and new clusters appeared in the anlage of the epiphysis
(18 hr) and in the pituitary and dorsal mesencephalon (by 24 hr).
Subsequently, as more neurons differentiated, the gaps of unlabeled cells
were reduced; by 48 hr, the cluster boundaries were indistinguishable.
Axonogenesis also followed a stereotyped sequence. The first HNK-1-labeled
processes arose from the first three clusters of AChE-positive cells and
connected the clusters. The earliest axonal growth cones appeared at 16 hr,
directed caudally from two to three neurons of the ventrocaudal cluster and
pioneering the ventral longitudinal tract. By 18 hr, the tract of the
postoptic commissure was initiated by growth cones directed caudally from
the ventrorostral cluster toward the ventrocaudal cluster. By 20 hr, axons
from the dorsorostral cluster projected ventrally to form the supraoptic
tract. The other dorsoventral tracts (the dorsoventral diencephalic tract
and the tract of the posterior commissure) became evident between 20 and 24
hr. These observations provide a continuous record of the topological
distortions involved in the conversion of the tubular embryonic brain into
the contorted adult form. The telencephalon, ventral diencephalon, and
hypothalamus originate from the same rostrocaudal level of the neural tube.
The pattern of differentiation demonstrated that the early development of
the rostral neural tube occurs simultaneously in several independent
centers, similar to the overtly segmental development of the hindbrain.
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