DEV02854 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. GFP immunostaining of E16.5 Cdk5^+/+ and Cdk5^−/− coronal sections that were electroporated with the CAG-EGFP plasmid at E14.5 in utero. CAG-EGFP-labelled (green) bipolar progenitors, which have a radial glial morphology and extend their fibres towards the pia, and multipolar migrating neurons in the premigratory zone are evident (arrows). Scale bar: 100 μm.

• Supplemental Figure S2 -

  Fig. S2. Tuj1-staining of Cdk5^+/+ and Cdk5^−/− neurons. (A) Tuj1-stained cultured cortical neurons at 3 DIV from Cdk5^+/+ and Cdk5^−/− embryos at E14.5. Pyramidal neurons have a single dendrite that gradually tapers off from the cell body (arrow), and non-pyramidal neurons have multiple dendrites (arrowheads). Scale bar: 100 μm. (B) No difference was observed between Cdk5^+/+ and Cdk5^−/− embryos (n=5) in the percentage of Tuj1-positive cells with the pyramidal morphology at 3 DIV.

• Supplemental Figure S3 -

  Fig. S3. Retarded growth and limited posterior extension of the cerebral cortex in CxCdk5KO mice. (A) CxCdk5KO mice had significantly lower body weight from P4. The body weights of CxCdk5KO mice and their littermate controls (n=6) were measured at the indicated postnatal day mean±s.d. (g). At P0, no significant difference was observed. However, lower body weights were evident from P4 until their death. *, P<0.01; **, P<0.05. (B) Gross examination of fixed brains revealed a smaller cerebral cortex in CxCdk5KO mice. Because of small body, overall brain sizes of CxCdk5KO mice were smaller than those of littermate controls. The proportion of the CxCdk5KO cerebral cortex was also small, and posterior extension (posterior margin is outlined by dotted lines) was limited. (C) Comparison of cerebral cortex in Nissl-stained sagittal sections revealed an abnormal histology in CxCdk5KO mice, including limited posterior extension of the cingulate cortex as indicated by the arrows.

• Supplemental Figure S4 -

  Fig. S4. Brain sections from YFP-H and reeler;YFP-H double-transgenic mice at P21. In the reeler;YFP-H mice, YFP-positive layer V neurons, which were scattered in the cerebral wall, preserve their pyramidal morphology despite their ectopic localisation. Arrows indicate cell bodies of YFP-positive layer V neurons and arrowheads indicate their apical dendrites. Directions of apical dendrite are abnormal in the ectopic layer V neurons as reported (Terashima et al., 1983).

• Movie 1 -

  Movie 1. An animated version of Fig. 3A. Recorded for 24 hours (30-minute intervals) from brain slices introduced with (A) CAG-EGFP or (B) CGC-Cdk5-DN. The transition from multipolar to bipolar shape and radial migration in the bipolar shape were observed in the slice with CAG-EGFP (A) but rarely seen in that with CGC-Cdk5-DN (B).

• Movie 2 -

  Movie 2. An animated version of Fig. 4A. Recorded for 48 hours (30-minute intervals) from (A) Cdk5^+/+ or (B) Cdk5^−/−embryos. Most Cdk5^+/+ neurons acquired bipolar shapes during the observation period and directed their leading processes to the pia (A). A few Cdk5^−/− neurons acquired the bipolar morphology, and some Cdk5^−/− neurons reached the pia in a multipolar shape (B). The majority of Cdk5^−/− neurons remained in the multipolar shape during the observation period (B).