Development • Supplementary information
Development 143: doi:10.1242/dev.134163: Supplementary information
Development 143: doi:10.1242/dev.134163: Supplementary information
Fig. S1. Morphological characteristics of transfected neurons in the intermediate zone five days
after electroporation at E14.5. (A) Control cells show characteristic features of migrating cortical
neurons with an oval soma, a thick leading process (white arrows) and a thin trailing process. (B)
CofilinWT-transfected cells preserved the asymmetric bipolar shape but their leading processes and cell
bodies gave rise to many small branches (white arrows). Small cofilin-actin rods were observed in these
branches (red arrowheads). (C) CofilinS3A-transfected cells displayed an asymmetric bipolar shape with
thick and short leading processes. Some neurons extended two or more leading processes (middle panel)
and gave rise to branches (white arrows). Small cofilin-actin rods were visible (red arrowheads). (D)
CofilinS3E-transfected neurons also preserved a bipolar shape but lost the characteristic asymmetric
polarity of migrating neurons. The two processes are equally short and thick (white arrows). No typical
leading or trailing processes could be recognized. (E) GFP-labeled reeler neurons displayed a variety of
different shapes with mal-oriented leading processes (white arrows). (F) Drawings of the leading
processes and branches of representative cortical neurons transfected with the various constructs by IUE.
Development • Supplementary information
Scale bar (A-E): 10 µm.
Development 143: doi:10.1242/dev.134163: Supplementary information
Movie 1. Migrating control neurons in a cortical slice culture three days after UE
long leading process pointing to the CP (top) and a trailing process descending to the
VZ (bottom). The neurons show directed migration towards the CP with leading
process elongation, forward movement of the cell body into the leading process, and
railing process retraction. Slice cultures were allowed to recover for about 3 hours
before imaging with an Improvision confocal spinning disc microscope was initiated
x20 air immersion objective). Acquired z-series were visualized as single optical
scans with concurrent orthogonal views using Velocity6 software. Duration of
imaging: about 6 hours; time interval: 10 min.
Development • Supplementary information
with pCAG-GFP. Neurons in the upper IZ exhibit a clear bipolar orientation with a
Development 143: doi:10.1242/dev.134163: Supplementary information
Movie 2. Migrating neurons in a cortical slice culture three days after IUE with
cofilinWT. The migrating neurons have retained a bipolar morphology in the upper IZ
(bottom) and CP (top). The migration behavior is not significantly different from that of
Development • Supplementary information
control cells. Imaging conditions were
we as described for Movie 1.
Development 143: doi:10.1242/dev.134163: Supplementary information
Movie 3. Migrating neurons in a cortical slice culture three days after IUE with
cofilinS3A. In the upper IZ (bottom), many neurons appear multipolar, and only a few cells
with very low plasmid expression have a bipolar shape and migrate normally towards the
CP (top). Within the CP, some cells give rise to leading processes
directed towards the VZ and also migrate in this direction. Imaging conditions were as
Development • Supplementary information
described for Movie 1.
Development 143: doi:10.1242/dev.134163: Supplementary information
Movie 4. Migrating neurons in a cortical slice culture three days after IUE with
cofilinS3E. The upper IZ and lower CP are crowded with numerous neurons showing a
thick, short process directed towards the VZ (bottom). Neuronal migration is
retarded resulting in cell-sparse upper and middle portions of the CP (top). Imaging
Development • Supplementary information
conditions were as described for Movie 1.
Development 143: doi:10.1242/dev.134163: Supplementary information
Movie 5. Migrating neurons in a reeler cortical slice culture three days after IUE
with pCAG-GFP. Neurons in the upper IZ (bottom) show a bipolar morphology with
their leading processes pointing to the CP. In contrast, many neurons in the CP (top)
give rise to a thick leading process directed towards the IZ. Their nuclei moved into
this process indicating a reversion of migration directionality. Imaging conditions were
Development • Supplementary information
as described for Movie 1.
Development 143: doi:10.1242/dev.134163: Supplementary information
Movie 6. Migrating neurons in a reeler cortical slice culture three days after IUE
with Limk1. When compared to reeler neurons transfected only with pCAG-GFP, 7
numerous neurons show normal migration directed towards the MZ (top). Imaging 8
Development • Supplementary information
conditions were as described for Movie 1.