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Journal of Neuroscience, Vol 12, 3257-3271, Copyright © 1992 by Society for Neuroscience
Dynamics of terminal arbor formation and target approach of retinotectal axons in living zebrafish embryos: a time-lapse study of single axons
RJ Kaethner and CA Stuermer
Faculty of Biology, University of Konstanz, Germany.
In a variety of species, developing retinal axons branch initially more
widely in their visual target centers and only gradually restrict their
terminal arbors to smaller and defined territories. Retinotectal axons in
fish, however, appeared to grow in a directed manner and to arborize only
at their retinotopic target sites. To visualize the dynamics of retinal
axon growth and arbor formation in fish, time-lapse recordings were made of
individual retinal ganglion cell axons in the tectum in live zebrafish
embryos. Axons were labeled with the fluorescent carbocyanine dyes Dil or
DiO inserted as crystals into defined regions of the retina, viewed with
40x and 100x objectives with an SIT camera, and recorded, with exposure
times of 200 msec at 30 or 60 sec intervals, over time periods of up to 13
hr. (1) Growth cones advanced rapidly, but the advance was punctuated by
periods of rest. During the rest periods, the growth cones broadened and
developed filopodia, but during extension they were more streamlined. (2)
Growth cones traveled unerringly into the direction of their retinotopic
targets without branching en route. At their target and only there, the
axons began to form terminal arborizations, a process that involved the
emission and retraction of numerous short side branches. The area that was
permanently occupied or touched by transient branches of the terminal
arbor--"the exploration field"--was small and almost circular and covered
not more than 5.3% of the entire tectal surface area, but represented up to
six times the size of the arbor at any one time. These findings are
consistent with the idea that retinal axons are guided to their retinotopic
target sites by sets of positional markers, with a graded distribution over
the axes of the tectum.
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