Fig. 3. Analysis of mitotic and post-mitotic specificity of recombination in the cerebral cortices of Emx1cre and Nestin-cre mice using the MADM-11 genetic.

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Fig. 3. Analysis of mitotic and post-mitotic specificity of recombination in the cerebral cortices of Emx1cre and Nestin-cre mice using the MADM-11 genetic system.(A) Scheme of cre-induced reporter expression in Mosaic Analysis with Double Markers on chromosome 11 (MADM-11) mice. Analysis of mitotic and post-mitotic specificity of recombination in the cerebral cortices of Emx1cre and Nestin-cre mice using the MADM-11 genetic system.(A) Scheme of cre-induced reporter expression in Mosaic Analysis with Double Markers on chromosome 11 (MADM-11) mice. In this line, cre-dependent mitotic recombination results in expression of GFP or tdTom independently or together yielding various cells with three potential expression patterns (tdTom+, red; GFP+, green; tdTom+/GFP+, yellow). Recombination can also occur in postmitotic cells during G0, but only yellow cells will be obtained due to the presence of only one allele for each reporter system. (B) MADM-11 recombined cells in E12.5 Nestin-cre and Emx1cre cerebral cortices. Only a few Nestin-cre:MADM11 cells were detected predominantly in the preplate (PPL) away from the VZ. Almost all recombined Nestin-cre:MADM-11 cells were negative for the mitotic marker PH3 (blue). In contrast, the Emx1cre knock-in allele was sufficient for driving mitotic recombination in the cerebral cortices at E12.5 as evidenced by the presence of cells expressing a single reporter. Co-localization of PH3 (blue) with recombined cells was detectable in the VZ (arrows). (C) Nestin-cre:MADM-11 recombined cells in E14.5 and E17.5 cerebral cortices. By E14.5, a few mitotically recombined cells were found in the IZ and CP, but very few clones could be detected in the VZ and SVZ. In addition, the majority of recombined cells in the VZ and SVZ were largely devoid of PH3 immunolabeling. By E17.5, mitotic recombination became sufficient in the entire cerebral cortex and PH3+ recombined cells could be readily detected in both the VZ and SVZ (arrows). Scale bars: B, 30 µm; C, 60 µm. Huixuan Liang et al. Biology Open 2012;bio.20122287 © 2012. Published by The Company of Biologists Ltd