Reinstating plasticity and memory in a tauopathy mouse model with an acetyltransferase activator
2018; Springer Nature; Volume: 10; Issue: 11 Linguagem: Inglês
10.15252/emmm.201708587
ISSN1757-4684
AutoresSnehajyoti Chatterjee, Raphaelle Cassel, Anne Schneider‐Anthony, Karine Mérienne, Brigitte Cosquer, Laura Tzeplaeff, Sarmistha Halder Sinha, Manoj Kumar, Piyush Chaturbedy, Muthusamy Eswaramoorthy, Stéphanie Le Gras, Céline Keime, Olivier Bousiges, P. Dutar, Petnoï Petsophonsakul, Claire Rampon, Jean‐Christophe Cassel, Luc Buée, David Blum, Tapas K. Kundu, Anne‐Laurence Boutillier,
Tópico(s)Neurogenesis and neuroplasticity mechanisms
ResumoResearch Article1 October 2018Open Access Source DataTransparent process Reinstating plasticity and memory in a tauopathy mouse model with an acetyltransferase activator Snehajyoti Chatterjee Snehajyoti Chatterjee orcid.org/0000-0002-4873-9416 Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France LNCA, CNRS UMR 7364, Strasbourg, France Search for more papers by this author Raphaelle Cassel Raphaelle Cassel Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France LNCA, CNRS UMR 7364, Strasbourg, France Search for more papers by this author Anne Schneider-Anthony Anne Schneider-Anthony Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France LNCA, CNRS UMR 7364, Strasbourg, France Search for more papers by this author Karine Merienne Karine Merienne Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France LNCA, CNRS UMR 7364, Strasbourg, France Search for more papers by this author Brigitte Cosquer Brigitte Cosquer Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France LNCA, CNRS UMR 7364, Strasbourg, France Search for more papers by this author Laura Tzeplaeff Laura Tzeplaeff Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France LNCA, CNRS UMR 7364, Strasbourg, France Search for more papers by this author Sarmistha Halder Sinha Sarmistha Halder Sinha Transcription and Disease Laboratory, Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, India Search for more papers by this author Manoj Kumar Manoj Kumar orcid.org/0000-0002-6838-2995 Transcription and Disease Laboratory, Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, India Search for more papers by this author Piyush Chaturbedy Piyush Chaturbedy Chemistry and Physics of Materials Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, India Search for more papers by this author Muthusamy Eswaramoorthy Muthusamy Eswaramoorthy Chemistry and Physics of Materials Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, India Search for more papers by this author Stéphanie Le Gras Stéphanie Le Gras CNRS, Inserm, UMR 7104, Microarray and Sequencing Platform, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg, Illkirch, France Search for more papers by this author Céline Keime Céline Keime CNRS, Inserm, UMR 7104, Microarray and Sequencing Platform, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg, Illkirch, France Search for more papers by this author Olivier Bousiges Olivier Bousiges Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France Laboratory of Biochemistry and Molecular Biology, Hôpital de Hautepierre, University Hospital of Strasbourg, Strasbourg, France Search for more papers by this author Patrick Dutar Patrick Dutar Centre de Psychiatrie et Neurosciences, INSERM UMRS894, Université Paris Descartes, Sorbonne Paris Cité, Paris, France Search for more papers by this author Petnoi Petsophonsakul Petnoi Petsophonsakul Centre de Recherches sur la Cognition Animale, Centre de Biologie Intégrative, CNRS, UPS, Université de Toulouse, Toulouse, France Search for more papers by this author Claire Rampon Claire Rampon Centre de Recherches sur la Cognition Animale, Centre de Biologie Intégrative, CNRS, UPS, Université de Toulouse, Toulouse, France Search for more papers by this author Jean-Christophe Cassel Jean-Christophe Cassel Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France LNCA, CNRS UMR 7364, Strasbourg, France Search for more papers by this author Luc Buée Luc Buée Inserm, CHU-Lille, UMR-S 1172, Alzheimer & Tauopathies, Université de Lille, Lille, France Search for more papers by this author David Blum David Blum orcid.org/0000-0001-5691-431X Inserm, CHU-Lille, UMR-S 1172, Alzheimer & Tauopathies, Université de Lille, Lille, France Search for more papers by this author Tapas K Kundu Corresponding Author Tapas K Kundu [email protected] orcid.org/0000-0001-7790-214X Transcription and Disease Laboratory, Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, India Search for more papers by this author Anne-Laurence Boutillier Corresponding Author Anne-Laurence Boutillier [email protected] orcid.org/0000-0002-2317-9280 Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France LNCA, CNRS UMR 7364, Strasbourg, France Search for more papers by this author Snehajyoti Chatterjee Snehajyoti Chatterjee orcid.org/0000-0002-4873-9416 Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France LNCA, CNRS UMR 7364, Strasbourg, France Search for more papers by this author Raphaelle Cassel Raphaelle Cassel Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France LNCA, CNRS UMR 7364, Strasbourg, France Search for more papers by this author Anne Schneider-Anthony Anne Schneider-Anthony Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France LNCA, CNRS UMR 7364, Strasbourg, France Search for more papers by this author Karine Merienne Karine Merienne Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France LNCA, CNRS UMR 7364, Strasbourg, France Search for more papers by this author Brigitte Cosquer Brigitte Cosquer Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France LNCA, CNRS UMR 7364, Strasbourg, France Search for more papers by this author Laura Tzeplaeff Laura Tzeplaeff Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France LNCA, CNRS UMR 7364, Strasbourg, France Search for more papers by this author Sarmistha Halder Sinha Sarmistha Halder Sinha Transcription and Disease Laboratory, Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, India Search for more papers by this author Manoj Kumar Manoj Kumar orcid.org/0000-0002-6838-2995 Transcription and Disease Laboratory, Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, India Search for more papers by this author Piyush Chaturbedy Piyush Chaturbedy Chemistry and Physics of Materials Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, India Search for more papers by this author Muthusamy Eswaramoorthy Muthusamy Eswaramoorthy Chemistry and Physics of Materials Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, India Search for more papers by this author Stéphanie Le Gras Stéphanie Le Gras CNRS, Inserm, UMR 7104, Microarray and Sequencing Platform, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg, Illkirch, France Search for more papers by this author Céline Keime Céline Keime CNRS, Inserm, UMR 7104, Microarray and Sequencing Platform, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg, Illkirch, France Search for more papers by this author Olivier Bousiges Olivier Bousiges Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France Laboratory of Biochemistry and Molecular Biology, Hôpital de Hautepierre, University Hospital of Strasbourg, Strasbourg, France Search for more papers by this author Patrick Dutar Patrick Dutar Centre de Psychiatrie et Neurosciences, INSERM UMRS894, Université Paris Descartes, Sorbonne Paris Cité, Paris, France Search for more papers by this author Petnoi Petsophonsakul Petnoi Petsophonsakul Centre de Recherches sur la Cognition Animale, Centre de Biologie Intégrative, CNRS, UPS, Université de Toulouse, Toulouse, France Search for more papers by this author Claire Rampon Claire Rampon Centre de Recherches sur la Cognition Animale, Centre de Biologie Intégrative, CNRS, UPS, Université de Toulouse, Toulouse, France Search for more papers by this author Jean-Christophe Cassel Jean-Christophe Cassel Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France LNCA, CNRS UMR 7364, Strasbourg, France Search for more papers by this author Luc Buée Luc Buée Inserm, CHU-Lille, UMR-S 1172, Alzheimer & Tauopathies, Université de Lille, Lille, France Search for more papers by this author David Blum David Blum orcid.org/0000-0001-5691-431X Inserm, CHU-Lille, UMR-S 1172, Alzheimer & Tauopathies, Université de Lille, Lille, France Search for more papers by this author Tapas K Kundu Corresponding Author Tapas K Kundu [email protected] orcid.org/0000-0001-7790-214X Transcription and Disease Laboratory, Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, India Search for more papers by this author Anne-Laurence Boutillier Corresponding Author Anne-Laurence Boutillier la[email protected] orcid.org/0000-0002-2317-9280 Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France LNCA, CNRS UMR 7364, Strasbourg, France Search for more papers by this author Author Information Snehajyoti Chatterjee1,2,‡, Raphaelle Cassel1,2,‡, Anne Schneider-Anthony1,2,‡, Karine Merienne1,2, Brigitte Cosquer1,2, Laura Tzeplaeff1,2, Sarmistha Halder Sinha3, Manoj Kumar3, Piyush Chaturbedy4, Muthusamy Eswaramoorthy4, Stéphanie Le Gras5, Céline Keime5, Olivier Bousiges1,6, Patrick Dutar7, Petnoi Petsophonsakul8, Claire Rampon8, Jean-Christophe Cassel1,2, Luc Buée9, David Blum9, Tapas K Kundu *,3 and Anne-Laurence Boutillier *,1,2 1Laboratoire de Neuroscience Cognitives et Adaptatives (LNCA), Université de Strasbourg, Strasbourg, France 2LNCA, CNRS UMR 7364, Strasbourg, France 3Transcription and Disease Laboratory, Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, India 4Chemistry and Physics of Materials Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, India 5CNRS, Inserm, UMR 7104, Microarray and Sequencing Platform, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg, Illkirch, France 6Laboratory of Biochemistry and Molecular Biology, Hôpital de Hautepierre, University Hospital of Strasbourg, Strasbourg, France 7Centre de Psychiatrie et Neurosciences, INSERM UMRS894, Université Paris Descartes, Sorbonne Paris Cité, Paris, France 8Centre de Recherches sur la Cognition Animale, Centre de Biologie Intégrative, CNRS, UPS, Université de Toulouse, Toulouse, France 9Inserm, CHU-Lille, UMR-S 1172, Alzheimer & Tauopathies, Université de Lille, Lille, France ‡These authors contributed equally to this work *Corresponding author. Tel: +91 80 2208 2840; E-mail: [email protected] *Corresponding author. Tel: +33 3688 51934; E-mail: [email protected] EMBO Mol Med (2018)10:e8587https://doi.org/10.15252/emmm.201708587 PDFDownload PDF of article text and main figures. Peer ReviewDownload a summary of the editorial decision process including editorial decision letters, reviewer comments and author responses to feedback. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Abstract Chromatin acetylation, a critical regulator of synaptic plasticity and memory processes, is thought to be altered in neurodegenerative diseases. Here, we demonstrate that spatial memory and plasticity (LTD, dendritic spine formation) deficits can be restored in a mouse model of tauopathy following treatment with CSP-TTK21, a small-molecule activator of CBP/p300 histone acetyltransferases (HAT). At the transcriptional level, CSP-TTK21 re-established half of the hippocampal transcriptome in learning mice, likely through increased expression of neuronal activity genes and memory enhancers. At the epigenomic level, the hippocampus of tauopathic mice showed a significant decrease in H2B but not H3K27 acetylation levels, both marks co-localizing at TSS and CBP enhancers. Importantly, CSP-TTK21 treatment increased H2B acetylation levels at decreased peaks, CBP enhancers, and TSS, including genes associated with plasticity and neuronal functions, overall providing a 95% rescue of the H2B acetylome in tauopathic mice. This study is the first to provide in vivo proof-of-concept evidence that CBP/p300 HAT activation efficiently reverses epigenetic, transcriptional, synaptic plasticity, and behavioral deficits associated with Alzheimer's disease lesions in mice. Synopsis Epigenetic dysfunctions related to H2B histone acetylation were shown in the hippocampus of a mouse model of Alzheimer's disease. A therapeutic strategy aimed at activating CBP/p300 acetyltransferase revealed efficient at rescuing neuronal activity, plasticity and memory in these mice. Alzheimer-related tau pathology is associated with epigenetic dysfunctions. A balanced H2Bac epigenomic landscape is maintained by CBP/HAT activity and important for hippocampus-dependent learning and memory. A new therapeutic strategy to restore memory in cognitive diseases can be achieved by pharmacological activation of CBP/p300 acetyltransferases with CSP-TTK21. Introduction Alzheimer's disease (AD) is characterized by the accumulation of amyloid beta peptides and abnormally phosphorylated Tau proteins, and a progressive impairment of plasticity and memory functions, ending in massive neuronal loss and dementia (Serrano-Pozo et al, 2011). Available treatments have minimal or no effect on the course of the disease, and most advanced strategies targeting the pathological hallmarks of AD (e.g., Aβ immunotherapies) have been recently proven unsuccessful (Abbott & Dolgin, 2016; The Lancet, 2017). Alternative strategies aim to restore neural circuits and plasticity (Canter et al, 2016). Epigenetic changes have emerged as important contributors of neurodegenerative diseases (Schneider et al, 2013; Coppede, 2014; Francelle et al, 2017), including AD (Fischer, 2014; Bennett et al, 2015). Histone acetylation has been particularly studied since it is an important regulator of plasticity and memory formation (Peixoto & Abel, 2013; Zovkic et al, 2013). Altered acetylation regulations are thus presumably involved in cognitive deficits. For instance, increased histone deacetylase 2 (HDAC2) levels leading to decreased histone acetylation have been observed at genes that are important for learning and memory, both in mouse models of AD and in post-mortem brains from patients with early-stage AD (Graff et al, 2012). Hence, several histone deacetylase (HDAC) inhibitors have been tested in different AD mouse models and some showed significant improvement in plasticity and memory functions (reviewed in Fischer, 2014; Graff & Tsai, 2013). Yet, few genome-wide-scale approaches have been performed using brain tissues from animal models or patients affected by neurodegenerative diseases, with pioneering work coming from studies of Huntington's disease (e.g., Vashishtha et al, 2013; Achour et al, 2015; Bai et al, 2015; Guiretti et al, 2016), and only two studies in AD models (Benito et al, 2015; Gjoneska et al, 2015). Tsai et al performed a profiling of transcriptional and epigenomic changes in the hippocampus of a mouse model of AD and showed reduction in H3K27 acetylation (H3K27ac) at downregulated genes which were enriched in synaptic plasticity genes, as well as increased H3K27ac at immune response-enriched upregulated genes (Gjoneska et al, 2015). Fischer et al tested the effects of the HDAC inhibitor suberoylanilide hydroxamic acid (SAHA) and showed it could reinstate physiological exon usage associated with H4K12 acetylation and plasticity gene expression in aged neurons, but this was not found in AD neurons, where the beneficial effect of SAHA was only partial (Benito et al, 2015). Thus, mechanisms underlying epigenetic changes in AD remain cryptic and the mode of action of potential epigenetic drugs and their consequences in AD remain to be established. Herein, using genome-wide-scale approaches (ChIP-seq/RNA-seq studies), we have investigated the molecular mechanisms associated with plasticity dysfunctions in the hippocampus of a mouse model with AD-like Tau pathology (THY-Tau22). This mouse strain exhibits neurofibrillary tangles (NFTs), neuroinflammation (Laurent et al, 2017), spatial memory deficits (Schindowski et al, 2006), impaired long-term depression (LTD; Van der Jeugd et al, 2011; Ahmed et al, 2015), and altered dendritic spine formation in the hippocampus (Burlot et al, 2015). In parallel, we have tested the potential therapeutic application of a newly developed CBP/p300 histone acetyltransferases (HAT) activator molecule (CSP-TTK21), as we recently showed that this molecule was able to acetylate nuclear chromatin in the mouse brain (Chatterjee et al, 2013). Indeed, as an alternative to HDAC inhibitors, targeting CBP/p300 HATs has been proposed as an innovative therapeutic strategy for disorders affecting memory (Schneider et al, 2013; Valor et al, 2013). Indeed, CBP loss of function has been reported during neurodegeneration (Rouaux et al, 2003, 2004) and CBP/p300 HATs play important roles in neuronal plasticity and cognition, including hippocampal long-term potentiation (LTP) and long-term memory (Alarcon et al, 2004; Vecsey et al, 2007; Barrett et al, 2011; Valor et al, 2011; West & Greenberg, 2011). In this study, we found that treatment of 8-month-old THY-Tau22 mice with CSP-TTK21 restored spatial memory and plasticity functions. At this age, the hippocampal transcriptome induced by spatial learning was severely impaired and CSP-TTK21 treatment re-established the expression of genes involved in neuronal plasticity (immediate early genes, IEGs) and of cognitive enhancers (e.g., Neurotensin, Klotho). Further, we found a specific decrease in H2B (H2Bac)—but not H3K27 (H3K27ac)—acetylation levels in the hippocampus of THY-Tau22 mice, and CBP/p300 activation with CSP-TTK21 significantly restored this signature, notably by a wide re-acetylation of H2B at transcription start sites (TSS) and CBP enhancers. Together, these data indicate that CBP/p300 HAT activation restores an epigenetic landscape that permits, either directly or indirectly, the upregulation of genes involved in neuronal plasticity (IEGs) and memory during learning, thus representing a promising therapeutic option for plasticity and memory enhancement. Results The CBP pathway is a potential therapeutic target in a mouse model of tauopathy In the THY-Tau22 mouse model, the tauopathy rapidly progresses once inflammation processes have started, around 7 months of age. Twelve-month-old THY-Tau22 mice exhibit strong tauopathy, with massive accumulation of abnormal Tau conformation and phosphorylation notably in the CA1 region of dorsal hippocampus (Schindowski et al, 2006). We found that neurons exhibiting pathological Tau hallmarks (AT100-positive neurons) showed depleted CBP protein levels (Fig 1A). Overall, we measured a global CBP reduction while neuronal marker NeuN levels remained unchanged, but this was associated with an increase in astrogliosis as expected at this age (Fig 1B). Several histone acetylation targets were evaluated by Western blot analyses, and we measured a decrease in H2B tetra-acetylation (H2BK5K12K15K20ac), whereas the global level of the other modifications tested was unchanged (Fig EV1A). In order to test the potential effect of our new drug, we carried out the study at 8 months of age, an earlier symptomatic age where mice already show memory deficits and inflammatory processes while pathology still progresses (Schindowski et al, 2006). Transcriptomic analyses performed in the dorsal hippocampus revealed moderate changes (51 dysregulated genes), but confirmed the presence of the inflammatory response signature (Fig 1C). In addition, a series of immediate early genes (IEGs) were downregulated, suggesting reduced basal neuronal activity in the hippocampus of THY-Tau22 vs. WT mice (Fig 1C). Lastly, predicted promoter motifs associated with the 15 downregulated genes were associated with cAMP pathway (ATF2/6 and CREB1; Fig 1C), which is a major regulator of IEGs. Thus, we aimed to test CSP-TTK21 molecule treatment as therapeutic intervention in these mice. Figure 1. Targeting the CBP/p300 pathway restores long-term spatial memory retention in THY-Tau22 mice A. Immunohistochemistry performed on 12-month-old THY-Tau22 mice showing AT100-positive neurons (green), CBP-positive nuclei (red), and DAPI-stained nuclei (blue) in the CA1 region of the dorsal hippocampus. A representative image is shown with a focus. Scale bars: 40 and 20 μm as noted (n = 5 mice). Arrows depict neurons bearing neurofibrillary tangles and do not display CBP immunostaining. The star depicts a ghost tangle (AT100-positive neuron and no nucleus). B. Western blot analyses performed in the dorsal hippocampus of 12-month-old THY-Tau22 mice compared to age-matched controls. NeuN, actin, tubulin, and total H2B levels are not changed. Phosphorylated Tau on serine 404 (Tau-Ph404) attests to samples from tauopathic mice. Quantification represents the ratio of the protein level detected on the total amount of proteins on the membranes, with WT arbitrarily set at 1 (fold induction). Bar graphs are mean ± SEM. n = 5–7/group as noted, multiple t-tests, and CBP *P = 0.003 and GFAP **P = 0.0001 for THY-Tau vs. WT mice. C. RNA-sequencing data performed in the hippocampus of 8-month-old mice. Functional analyses (DAVID GOTERM) performed on significantly deregulated genes in THY-Tau22 mice compared to WT mice. Representative genes are presented below. |log2 Fold Change| > 0.2; * indicates FDR < 0.05. IEG, immediate early genes. Predicted promoter motif (right) analyses performed with GREAT. Groups: WT mice (n = 3); THY-Tau22 mice (n = 2). Graphs are mean ± SEM. D. Long-term spatial memory testing: Mice were injected three times (1 per week) with vehicle (WT mice saline, WT VEH, n = 17), vehicle (THY-Tau22 mice CSP, 500 μg/mouse, TAU VEH, n = 10), or molecule (THY-Tau22 mice CSP-TTK21, 500 μg/mice TAU MOL, n = 13) before training of spatial memory in the Morris water maze (MWM); retention (Probe test) was tested 10 days after the last training session. Acquisition (escape latencies, seconds) and retention performances (time in target quadrant, seconds) are shown for the three groups of mice. All groups of mice displayed significant acquisition of the platform location [Day effect, F(4,148) = 26.45, P < 0.001; Group and Group × Day effects, ns], but only TAU VEH exhibited impaired retention. CSP-TTK21 treatment fully restored the ability of THY-Tau22 mice to remember the platform location. Bar graphs are mean ± SEM. Student's t-test to a constant value, # when compared to random (dotted line, 15 s) WT VEH, t(16) = 4.6323, #P = 0.0002; TAU VEH, t(9) = 0.3606, P = 0.7267; TAU MOL, t(12) = 3.945, #P = 0.0019. One-way ANOVA; F(2,37) = 3.55; P = 0.03; * in the different comparisons: TAU MOL vs. TAU VEH, *P = 0.0166; WT VEH vs. TAU VEH, *P = 0.040; TAU MOL vs. WT VEH, non-significant P = 0.437). TQ, target quadrant; O, other corresponds to the mean of the three other quadrants. Download figure Download PowerPoint Click here to expand this figure. Figure EV1. CSP-TTK21 treatment of THY-Tau22 mice improves long-term spatial memory retention and induces some histone acetylation in the hippocampus A. Western blot analyses of dorsal hippocampal extracts from 12-month-old THY-Tau22 vs. aged-matched WT controls. Levels of different histone modifications were tested. Only the tetra-acetylated form of H2B (K5K10K15K20) shows a significant decrease in tauopathic mice. Normalization was performed on the total amount of proteins transferred onto the membranes, and data are presented as fold induction of the acetylated histone/total H2B histone ratio. n = 5–6/group as noted. Multiple t-tests, and H2BK5K10K15K20, *P = 0.0047 for THY-Tau22 vs. WT mice. B. Scatter plot presenting RNA-seq data comparison between THY-Tau22 vs. WT mice. The log2(Fold-Change) was estimated by DESeq2. Red dots correspond to genes with adjusted P-value < 0.05. C. Habituation consisted of one trial to the visible platform. No statistical difference was observed in the distance from the habituation platform depending on the Group (F(1,37) = 0.17, P = 0.84). D. Distance to reach the platform (in meters) during the acquisition period. All groups of mice displayed significant and similar decreased of the distance traveled to reach the platform depending on the day [Day effect, F(4,148) = 26.47, P < 0.001; Group and Group × Day effects, ns]. Two-way ANOVA followed by a multiple-comparisons test (Newman–Keuls). E. The swim speed during the acquisition was the same for all groups [Day effect, (F(4,148) = 0.95, P = 0.46) and Group effect (F(2.37) = 1.06, P = 0.35)]. Two-way ANOVA followed by a multiple-comparisons test (Newman–Keuls). F. The number of visit to the previous platform location showed a significant difference in the number of crossing performed into it, and the three others for WT VEH mice (t(17) = 2.62, P < 0.05) and TAU MOL (t(13) = 2.70, P < 0.05) while there was no difference for the TAU VEH (t(10) = 0.21, P = 0.83). One-way ANOVA, Student's t-test, # P < 0.05 vs. mean of three others. G. The latency to first visit to the target platform compared to the mean latencies to the first visit to three others platform locations during the probe trial highlighted a global difference [Group Effect (F(2.37) = 5.87, P = 0.008) due to TAU VEH mice that significantly differed from the two others groups (TAU VEH vs. WT VEH (P = 0.004) and vs. TAU MOL (P = 0.012)]. There was a significant difference between the latencies to the first visit to the target platform with the three others for WT mice (t(17) = 6.90, P < 0.001) and TAU MOL (t(13) = 2.30, P < 0.05) while there was no difference for the TAU MOL (t(10) = 0.82, P = 0.42). One-way ANOVA, post hoc analyses *P < 0.05 vs. TAU VEH and Student's t-test, #P < 0.05 vs. mean of three others. H. Tracks representing the platform search during retention is shown for one mouse from each group that was either closest to the mean track or best of the group. The result of the probe test is noted below (in seconds). I. Western blot analyses were performed 22 days post-injection from experiment shown in Fig 1D, on n = 5 mice/group. Levels of the different histone modifications were tested. Normalization was performed on the total amount of proteins transferred onto the membranes and data are presented as fold induction of the acetylated histone/total H2B histone ratio. Multiple t-tests, and H2BK5K10K15K20ac, *P = 0.0006 and H3K27ac, *P = 0.0054 for CSP vs. CSP-TTK21, (#) indicates a tendency (P = 0.0736). Data information: Graphs are mean ± SEM. (C–G) Analyses of different parameters on the mice used in the Morris water maze experiment described in Fig 1 (WT VEH, n = 17, TAU VEH, n = 10 and TAU MOL, n = 13). Download figure Download PowerPoint Spatial reference memory tested in the Morris water maze (MWM) is a hippocampus-dependent memory task, in which 8-month-old THY-Tau22 mice fail when the retention test is delayed (Fig 1D). We injected mice three times with CSP-TTK21 (TAU MOL) or the control vehicle (TAU VEH) prior to spatial training (5-day acquisition) and tested mice at a 10-day post-training delay. WT littermates injected with saline vehicle (WT VEH) served as learning control. All mice showed similar acquisition performances in escape latencies (Fig 1D) or distance to the platform (Fig EV1D). Other parameters were also identical (habituation cued trial, swim speed; Fig EV1C and E). However, TAU VEH mice showed a clear retention deficit in the probe test (WT VEH: 23.4 vs. TAU VEH: 15.9 s in the target quadrant), which was prevented when the THY-Tau22 mice received CSP-TTK21 treatment before training (TAU MOL: 23.4 vs. TAU VEH: 15.9 s in the target quadrant; Fig 1D). CSP-TTK21-treated THY-Tau22 mice were also more precise when assessing the latency to first visit to the target quadrant (Fig EV1F) and the search strategy (see Closest to mean and Best tracks Fig EV1H). Only a tendency was measured when assessing platform crossing (Fig EV1G). Thus, CSP-TTK21 treatment fully restored the ability to form long-term memory in the THY-Tau22 mice. Lastly, H2BK5K12K15K20ac and H3K27ac were still more elevated in TAU MOL vs. TAU VEH in the hippocampus of mice euthanized 1 week after the probe test (22 days post-injection, Fig EV1I). Activation of the CBP/p300 HAT pathway restores synaptic plasticity in the hippocampus of THY-Tau22 mice We next determined whether CSP-TTK21 treatment could affect structural plasticity such as dendritic spine formation. Spines can be identified based on their morphological appearance. Spines with no head such as filopodia are considered immature. Stubby spines show a protrusion but no head or neck and are less mature than headed spines (thins and mushrooms) according to Harris et al (1992). Mushroom spines are thought to be stabilized by learning processes to form new synapses (Restivo et al, 2009; Caroni et al, 2014). We first tested the effe
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