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Fig. 5. Distribution and dynamics of GFP-synaptobrevin labeled presynaptic sites
along RGC axon terminals. (A) Time-lapse sequence of a region of a control
arbor illustrates the dynamic relationship between presynaptic site location
and axon branch formation. New axonal branches originate from sites rich in
GFP-synaptobrevin puncta (arrowheads), while new GFP-synaptobrevin clusters
appear along an axon branch (Alsina et al.,
2001). (B) Magnified region of an arbor illustrates the
localization of GFP-synaptobrevin puncta to a nascent branch (arrows) in a
DsRed2 labeled axon and its disappearance after anti-BDNF treatment (overlay,
top panel; GFP-synaptobrevin fluorescence only, bottom panel). In some
branches, GFP-synaptobrevin cluster dismantling precedes axon branch
elimination (arrow), as indicated by the significant decrease in GFP
fluorescence at the 2 hour time point. (C) Time-lapse sequence of a region of
an anti-BDNF treated axon arbor shows the disappearance of GFP-synaptobrevin
clusters and the retraction of an axon branch (arrow). The arrowhead indicates
a site where a decrease in punctuate GFP-synaptobrevin fluorescence correlates
with the shortening of the distal region of the axon branch. Asterisks
indicate arbor sites with stable GFP-synaptobrevin clusters. Scale bars: 20
µm in A; 10 µm in B,C. Posterior is upwards, anterior is downwards.
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