The Journal of Neuroscience, September 13, 2006, ():
High Levels of Cre Expression in Neuronal Progenitors Cause Defects in Brain Development Leading to Microencephaly and Hydrocephaly
J. Neurosci. Forni et al.
26: 9593
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplementary Fig. 1
MBP staining (A,D), DAPI staining (B,E), and DAPI and MBP merge (C,F) of fronto-parietal coronal sections at P18 of (A-C) wild type and (D-F) homozygous Bal1 Nestin Cre littermates show reduction and disorganization of myelinated axons and lower cell density. CC: corpus callosum Scale bar=100 μm
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Supplementary Fig. 2
Immunostaining for Nestin (cytoplasmic, red) and Cre recombinase (nuclear, green), merge (red and green) in heterozygous Bal1 Nestin Cre pups at P6 reveals expression of both proteins in (A-C) the SVZ with sustained Cre nuclear expression throughout the upper layers of the cortex. (D-F) Co-localization is better observed in the ependyma. (G-I) The subgranular zone (SGZ) of the hippocampus dentate gyrus reveals increasing levels of Cre nuclear positivity in post-mitotic cells relative to those in the proliferative zone. (L-N) A similar pattern is observed in the developing cerebellum where higher levels of nuclear staining are found in the inner granule layer (IGL). Scale bars=100μm (A-C) and 50μm (D-N)
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Supplementary Fig. 3
Western Blot analysis of whole brain (P1) reveals higher levels of Cre recombinase expression in Bal1 Nestin Cre homozygous mice (Bal1/Bal1) compared to Bal1 heterozygous (Bal1/WT) littermates; wild type brain (WT) was used as negative control. Tubulin and p21Cip-1 were used as loading controls.
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Supplementary Fig. 4
Quantification of ventricular dilation in E 19.5 heterozygous Line 1 and 4 NestinCreERT2 embryos treated with the indicated tamoxifen dosage. The extent of ventricular dilation is expressed as % of brain area occupied by the ventricles. Measurements were done as indicated in Materials and Methods. Bars represent mean + s.d.; n= 3 for each genotype.
t-test:*p<0.001.
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