Movie
1. Transport
of EGFP-SMN granules within neuronal processes. Cultured chick forebrain
neurons were transfected with EGFP-SMN and imaged live using a cooled
CCD camera. Shown here is a time lapse that depicts bi-directional
movements of EGFP-SMN granules (shown in black). One can see persistent
movements of granules in both anterograde and retrograde directions.
One small granule (white bar) takes a rapid retrograde trajectory
in the primary neurite toward a branch point and then moves in an
anterograde direction into a secondary neurite. Large granules often
displayed either oscillatory or bi-directional movements over short
distances. Note that the GFP signal is inverted and appears black.
(see Figure 3A)
Movie 2. Movements of EGFP-SMNDEx7 aggregates within
nucleus. Cultured chick forebrain neurons were transfected with the
exon-7 deletion mutant (EGFP-SMNDEx7) and imaged live using a cooled
CCD camera. Shown here is a time lapse that depicts an exclusive nuclear
localization in the form of aggregates. There was no detection of
EGFP-SMNDEx7 granules in the perinuclear cytoplasm. (see
Figure 6E)
Movie 3. The GAP-43 membrane targeting sequence directs
EGFP-SMNDEx7 granules into neuronal processes. Cultured chick forebrain
neurons were transfected with mem/EGFP-SMNDEx7 and imaged live using
a cooled CCD camera. This time lapse demonstrates bi-directional granule
movements characteristic of the full length EGFP-SMN. One granule
is observed to take a long distance anterograde trajectory in an axon-like
process that was followed by a shorter retrograde trajectory (white
bar). (see
Figure 8C)
Movie 4. Active transport of mem/EGFP-SMNDEx7 granules
from the cell body into the proximal neurite. Cultured chick forebrain
neurons were transfected with the exon-7 deletion mutant fused to
the GAP43 targeting sequence (mem/EGFP-SMNDEx7) and imaged live using
a cooled CCD camera. This time lapse demonstrate the movement of granule
(black) from the cytoplasm into the proximal neurite (white bar).
(see
Figure 8C)