Abstract
Mutations in the gene encoding the transcriptional modulator methyl-CpG binding protein 2 (MeCP2) are responsible for the neurodevelopmental disorder Rett syndrome which is one of the most frequent sources of intellectual disability in women. Recent studies showed that loss of Mecp2 in astrocytes contributes to Rett-like symptoms and restoration of Mecp2 can rescue some of these defects. The goal of this work is to compare gene expression profiles of wild-type and mutant astrocytes from Mecp2308/y mice (B6.129S-MeCP2<tm1Heto>/J) by using Affymetrix mouse 2.0 microarrays. Results were confirmed by quantitative real-time RT-PCR and by Western blot analysis. Gene set enrichment analysis utilizing Ingenuity Pathways was employed to identify pathways disrupted by Mecp2 deficiency. A total of 2152 genes were statistically differentially expressed between wild-type and mutated samples, including 1784 coding transcripts. However, only 257 showed fold changes >1.2. We confirmed our data by replicative studies in independent primary cultures of cortical astrocytes from Mecp2-deficient mice. Interestingly, two genes known to encode secreted proteins, chromogranin B and lipocalin-2, showed significant dysregulation. These proteins secreted from Mecp2-deficient glia may exert negative non-cell autonomous effects on neuronal properties, including dendritic morphology. Moreover, transcriptional profiling revealed altered Nr2f2 expression which may explain down- and upregulation of several target genes in astrocytes such as Ccl2, Lcn2 and Chgb. Unraveling Nr2f2 involvement in Mecp2-deficient astrocytes could pave the way for a better understanding of Rett syndrome pathophysiology and offers new therapeutic perspectives.
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Acknowledgments
This work was supported by Institut National de la Santé et de Recherche Médicale, and the Association Française du Syndrome de Rett (AFSR), and Fondation Jerome Lejeune (FJL R12212KK). CD is supported by the LabEX “Who Am I?” (PRES Sorbonne Paris Cité, Paris Diderot, Paris Descartes, CNRS, INSERM). We thank Florent Dumont at the sequencing platform and the staff of the animal facilities (Institut Cochin, Paris, France).
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Table S1
List of 257 dysregulated genes with at least a 1.20-fold expression variation among the 2152 coding and uncoding genes with significantly different expression between wild-type and Mecp2-deficient astrocytes. 130 genes were overexpressed and 127 were underexpressed in mutant astrocytes compared to wild-type astrocytes. p values and absolute expression values are indicated. (XLS 103 kb)
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Delépine, C., Nectoux, J., Letourneur, F. et al. Astrocyte Transcriptome from the Mecp2308-Truncated Mouse Model of Rett Syndrome. Neuromol Med 17, 353–363 (2015). https://doi.org/10.1007/s12017-015-8363-9
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DOI: https://doi.org/10.1007/s12017-015-8363-9