5a4ff0b3d777abf2a9f1973f9e1dcab7 Global_Impairment_of_Immediate-Early.pdf c6dd1551fe1db41d1ed98756436adac7c4fd67aa Global_Impairment_of_Immediate-Early.pdf d6520fa60f5855c620dc10ae300bd33f6d5be6bf71624c05176e92da741b496f Global_Impairment_of_Immediate-Early.pdf Title: Global Impairment of Immediate-Early Genes Expression in Rett Syndrome Models and Patients Linked toMyelination Defects Subject: Rett syndrome (RTT) is a severe neurodevelopmental disease caused almost exclusively by mutations to the MeCP2 gene. This disease may be regarded as a synaptopathy, with impairments affecting synaptic plasticity, inhibitory and excitatory transmission and network excitability. The complete understanding of the mechanisms behind how the transcription factor MeCP2 so profoundly affects the mammalian brain are yet to be determined. What is known, is that MeCP2 involvement in activity-dependent expression programs is a critical link between this protein and proper neuronal activity, which allows the correct maturation of connections in the brain. By using RNA-sequencing analysis, we found several immediate-early genes (IEGs, key mediators of activity-dependent responses) directly bound by MeCP2 at the chromatin level and upregulated in the hippocampus and prefrontal cortex of the Mecp2-KO mouse. Quantification of the IEGs response to stimulus both in vivo and in vitro detected an aberrant expression pattern in MeCP2-deficient neurons. Furthermore, altered IEGs levels were found in RTT patient’s peripheral blood and brain regions of post-mortem samples, correlating with impaired expression of downstream myelination-related genes. Altogether, these data indicate that proper IEGs expression is crucial for correct synaptic development and that MeCP2 has a key role in the regulation of IEGs. Keywords: Rett syndrome; MeCP2; EGR2; IEGs; myelination Author: Paolo Petazzi, Olga Caridad Jorge-Torres, Antonio Gomez, Iolanda Scognamiglio, Jordi Serra-Musach, Angelika Merkel, Daniela Grases, Clara Xiol, Mar O’Callaghan, Judith Armstrong, Manel Esteller and Sonia Guil Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Wed Jan 11 14:36:45 2023 CET ModDate: Wed Jan 11 14:40:09 2023 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 19 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2514574 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- TBJCMY+URWPalladioL-Bold Type 1 Custom yes yes no 10 0 XCEBDS+URWPalladioL-Roma Type 1 Custom yes yes no 15 0 TNZLVT+URWPalladioL-Ital Type 1 Custom yes yes no 19 0 UWJUYN+CMSY10 Type 1 Builtin yes yes no 23 0 IBTYZX+VnURWPalladioL Type 1 Custom yes yes no 27 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Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 735 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Results Destination: section.2 Children: Item: Title: RNA-Seq Analysis Reveals Dysregulation of IEGs in a Mouse Model of RTT Destination: subsection.2.1 Item: Title: MeCP2 Has a Potential Repressive Role on IEGs Destination: subsection.2.2 Item: Title: IEGs Response to Activation Is Misregulated in Mecp2-KO Neurons Destination: subsection.2.3 Item: Title: IEGs Are Over-Activated in Kainate-Stimulated Hippocampus In Vivo Destination: subsection.2.4 Item: Title: IEGs Are Dysregulated in Different Regions from Human RTT Post-Mortem Brains and Patient’s Peripheral Blood Destination: subsection.2.5 Item: Title: Aberrant EGR2 Expression Correlates with Altered Myelination Pathways in RTT Patients Destination: subsection.2.6 Item: Title: Discussion Destination: section.3 Item: Title: Materials and Methods Destination: section.4 Children: Item: Title: Animals Destination: subsection.4.1 Item: Title: Human Tissues Destination: subsection.4.2 Item: Title: Total RNA Isolation Destination: subsection.4.3 Item: Title: RNA-Seq Analysis Destination: subsection.4.4 Item: Title: Quantitative Real-Time PCR Destination: subsection.4.5 Item: Title: Chromatin Immunoprecipitation Destination: subsection.4.6 Item: Title: Chromatin Accessibility Assay Destination: subsection.4.7 Item: Title: Primary Neuronal Hippocampal and Cortical Cultures Destination: subsection.4.8 Item: Title: In Vivo Kainic Acid Administration Destination: subsection.4.9 Item: Title: Western Blot Destination: subsection.4.10 Item: Title: Brain Tissue Immunofluorescence Destination: subsection.4.11 Item: Title: Statistical Analysis Destination: subsection.4.12 Item: Title: References Destination: subsection.4.12 Info: Title: Global Impairment of Immediate-Early Genes Expression in Rett Syndrome Models and Patients Linked toMyelination Defects Author: Paolo Petazzi, Olga Caridad Jorge-Torres, Antonio Gomez, Iolanda Scognamiglio, Jordi Serra-Musach, Angelika Merkel, Daniela Grases, Clara Xiol, Mar O’Callaghan, Judith Armstrong, Manel Esteller and Sonia Guil Subject: Rett syndrome (RTT) is a severe neurodevelopmental disease caused almost exclusively by mutations to the MeCP2 gene. This disease may be regarded as a synaptopathy, with impairments affecting synaptic plasticity, inhibitory and excitatory transmission and network excitability. The complete understanding of the mechanisms behind how the transcription factor MeCP2 so profoundly affects the mammalian brain are yet to be determined. What is known, is that MeCP2 involvement in activity-dependent expression programs is a critical link between this protein and proper neuronal activity, which allows the correct maturation of connections in the brain. By using RNA-sequencing analysis, we found several immediate-early genes (IEGs, key mediators of activity-dependent responses) directly bound by MeCP2 at the chromatin level and upregulated in the hippocampus and prefrontal cortex of the Mecp2-KO mouse. Quantification of the IEGs response to stimulus both in vivo and in vitro detected an aberrant expression pattern in MeCP2-deficient neurons. Furthermore, altered IEGs levels were found in RTT patient’s peripheral blood and brain regions of post-mortem samples, correlating with impaired expression of downstream myelination-related genes. Altogether, these data indicate that proper IEGs expression is crucial for correct synaptic development and that MeCP2 has a key role in the regulation of IEGs. 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253, 456 Annotation: Subtype: Link Rect: 132, 423, 170, 433 Annotation: Subtype: Link Rect: 117, 389, 155, 399 Annotation: Subtype: Link Rect: 439, 365, 476, 376 Annotation: Subtype: Link Rect: 465, 342, 503, 353 Annotation: Subtype: Link Rect: 509, 342, 545, 353 Annotation: Subtype: Link Rect: 354, 319, 391, 330 Annotation: Subtype: Link Rect: 397, 319, 433, 330 Annotation: Subtype: Link Rect: 361, 284, 398, 295 Annotation: Subtype: Link Rect: 405, 284, 441, 295 Checksum: 1de9f1b4 Type: CRC32 Checksum: 5a4ff0b3d777abf2a9f1973f9e1dcab7 Type: MD5 Checksum: c6dd1551fe1db41d1ed98756436adac7c4fd67aa Type: SHA-1 Checksum: d6520fa60f5855c620dc10ae300bd33f6d5be6bf71624c05176e92da741b496f Type: SHA-256