Proteomic Analysis of Post-synaptic Density Fractions from Shank3 Mutant Mice Reveals Brain Region Specific Changes Relevant to Autism Spectrum Disorder

Reim, Dominik; Distler, Ute; Halbedl, Sonja; Verpelli, Chiara; Sala, Carlo; Bockmann, Juergen; Tenzer, Stefan; Boeckers, Tobias M.; Schmeißer, Michael J.

doi:10.3389/fnmol.2017.00026

Cited by 68 publications

(88 citation statements)

References 54 publications

(95 reference statements)

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“…In previously published models, analysis of anxiety-like behavior measured either elevated mazes, in the open fields or in dark/light emergence boxes have demonstrated a relationship between the targeted isoforms and the manifestations of anxiety like-behavior. While little differences were observed in mouse models with Δ4-9, Δ4-7, Δ9, and Δ11 deletions ( Peça et al, 2011 ; Wang et al, 2011 ; Schmeisser et al, 2012 ; Yang et al, 2012 ; Drapeau et al, 2014 ; Lee et al, 2015 ; Jaramillo et al, 2016 ; Reim et al, 2017 ; Vicidomini et al, 2017 ) increased levels of anxiety were reported in mice with Δ13, Δ13-16, and Δ21 deletions or point mutations ( Peça et al, 2011 ; Kouser et al, 2013 ; Speed et al, 2015 ; Mei et al, 2016 ; Zhou et al, 2016 ; Copping et al, 2017 ; Jaramillo et al, 2017 ). Increased levels of anxiety were confirmed in the light-dark emergence test and in the open field in the Δ4-22 mouse model from Wang et al, 2016b and colleagues and in the elevated maze and in the open field in our model.…”

Section: Discussionmentioning

confidence: 97%

Behavioral Phenotyping of an Improved Mouse Model of Phelan–McDermid Syndrome with a Complete Deletion of theShank3Gene

Drapeau

Riad

Kajiwara

et al. 2018

eNeuro

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Phelan–McDermid syndrome (PMS) is a rare genetic disorder in which one copy of the SHANK3 gene is missing or mutated, leading to a global developmental delay, intellectual disability (ID), and autism. Multiple intragenic promoters and alternatively spliced exons are responsible for the formation of numerous isoforms. Many genetically-modified mouse models of PMS have been generated but most disrupt only some of the isoforms. In contrast, the vast majority of known SHANK3 mutations found in patients involve deletions that disrupt all isoforms. Here, we report the production and thorough behavioral characterization of a new mouse model in which all Shank3 isoforms are disrupted. Domains and tasks examined in adults included measures of general health, neurological reflexes, motor abilities, sensory reactivity, social behavior, repetitive behaviors, cognition and behavioral inflexibility, and anxiety. Our mice are more severely affected than previously published models. While the deficits were typically more pronounced in homozygotes, an intermediate phenotype was observed for heterozygotes in many paradigms. As in other Shank3 mouse models, stereotypies, including increased grooming, were observed. Additionally, sensory alterations were detected in both neonatal and adult mice, and motor behavior was strongly altered, especially in the open field and rotarod locomotor tests. While social behaviors measured with the three-chambered social approach and male-female interaction tests were not strongly impacted, Shank3-deficient mice displayed a strong escape behavior and avoidance of inanimate objects in novel object recognition, repetitive novel object contact, marble burying, and nest building tasks, indicating increased novelty-induced anxiety. Similarly, increased freezing was observed during fear conditioning training and amygdala-dependent cued retrieval. Finally, deficits were observed in both initial training and reversal in the Barnes maze and in contextual fear testing, which are memory tasks involving hippocampal-prefrontal circuits. In contrast, working memory in the Y-maze spontaneous alternation test was not altered. This new mouse model of PMS, engineered to most closely represent human mutations, recapitulates core symptoms of PMS providing improvements for both construct and face validity, compared to previous models.

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Section: Discussionmentioning

confidence: 97%

Behavioral Phenotyping of an Improved Mouse Model of Phelan–McDermid Syndrome with a Complete Deletion of theShank3Gene

Drapeau

Riad

Kajiwara

et al. 2018

eNeuro

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“…A comprehensive analysis of mTORC1 activity in various brain regions of Shank3 mutant mice, together with the direct measurement of ribosomal quantity (both mRNA and protein) and activity, could provide important clues regarding the detailed mechanisms behind this. In this regard, it is worth noting that in a recent quantitative proteomic analysis of the striatal PSD of Shank3 mutant mice ( Shank3 Δ 11 −/− ), the levels of several ribosomal proteins were increased compared with WT mice ( Reim et al, 2017 ). Shank3 proteins are coupled with metabotropic glutamate receptors (mGluRs) via Homer ( Tu et al, 1999 ) and regulate their synaptic expression and signaling ( Verpelli et al, 2011 ; Vicidomini et al, 2016 ; Wang et al, 2016 ).…”

Section: Discussionmentioning

confidence: 99%

Integrative Brain Transcriptome Analysis Reveals Region-Specific and Broad Molecular Changes in Shank3-Overexpressing Mice

Jin

Kang

Ryu

et al. 2018

Front. Mol. Neurosci.

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Variants of the SH3 and multiple ankyrin repeat domain 3 (SHANK3) gene, encoding excitatory postsynaptic core scaffolding proteins, are causally associated with numerous neurodevelopmental and neuropsychiatric disorders, including autism spectrum disorder (ASD), bipolar disorder, intellectual disability, and schizophrenia (SCZ). Although detailed synaptic changes of various Shank3 mutant mice have been well characterized, broader downstream molecular changes, including direct and indirect changes, remain largely unknown. To address this issue, we performed a transcriptome analysis of the medial prefrontal cortex (mPFC) of adult Shank3-overexpressing transgenic (TG) mice, using an RNA-sequencing approach. We also re-analyzed previously reported RNA-sequencing results of the striatum of adult Shank3 TG mice and of the prefrontal cortex of juvenile Shank3+/ΔC mice with a 50–70% reduction of Shank3 proteins. We found that several myelin-related genes were significantly downregulated specifically in the mPFC, but not in the striatum or hippocampus, of adult Shank3 TG mice by comparing the differentially expressed genes (DEGs) of the analyses side by side. Moreover, we also found nine common DEGs between the mPFC and striatum of Shank3 TG mice, among which we further characterized ASD- and SCZ-associated G protein-coupled receptor 85 (Gpr85), encoding an orphan Gpr interacting with PSD-95. Unlike the mPFC-specific decrease of myelin-related genes, we found that the mRNA levels of Gpr85 increased in multiple brain regions of adult Shank3 TG mice, whereas the mRNA levels of its family members, Gpr27 and Gpr173, decreased in the cortex and striatum. Intriguingly, in cultured neurons, the mRNA levels of Gpr27, Gpr85, and Gpr173 were modulated by the neuronal activity. Furthermore, exogenously expressed GPR85 was co-localized with PSD-95 and Shank3 in cultured neurons and negatively regulated the number of excitatory synapses, suggesting its potential role in homeostatic regulation of excitatory synapses in Shank3 TG neurons. Finally, we performed a gene set enrichment analysis of the RNA-sequencing results, which suggested that Shank3 could affect the directional expression pattern of numerous ribosome-related genes in a dosage-dependent manner. To sum up, these results reveal previously unidentified brain region-specific and broad molecular changes in Shank3-overexpressing mice, further elucidating the complexity of the molecular pathophysiology of SHANK3-associated brain disorders.

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“…While many of the genes enriched for D1+ and D2+ expression are not exclusive to the striatum and are more broadly expressed (as demonstrated by the enrichment signal at the lowest specificity threshold), the striatum has been implicated in ID and autism pathology by numerous studies [97][98][99][100][101][102][103][104][105][106] . The striatum is particularly compelling as it has been linked to repetitive behaviors 97 core to the autism phenotype and also to genes known to be involved in DD, including CHD8, SHANK3, FOXP2, and KCNA4 98,99,102,105,106 . While the striatum is most strongly linked to autism core phenotypes, our observation of enrichment in a more general DD cohort suggests that, while the general bias of cortex genes to ID and striatum genes to ASD 101 still holds, the diverse expression patterns of genes across the brain at complex developmental time points may have substantial functional overlap among subtypes of NDDs that will require deep phenotyping and imaging to tease apart.…”

Section: Discussionmentioning

confidence: 99%

Neurodevelopmental disease genes implicated byde novomutation and CNV morbidity

Coe

Stessman²,

Sulovari

et al. 2017

Preprint

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We combined de novo mutation (DNM) data from 10,927 cases of developmental delay and autism to identify 301 candidate neurodevelopmental disease genes showing an excess of missense and/or likely gene-disruptive (LGD) mutations. 164 genes were predicted by two different DNM models, including 116 genes with an excess of LGD mutations. Among the 301 genes, 76% show DNM in both autism and intellectual disability/developmental delay cohorts where they occur in 10.3% and 28.4% of the cases, respectively. Intersecting these results with copy number variation (CNV) morbidity data identifies a significant enrichment for the intersection of our gene set and genomic disorder regions (36/301, LR+ 2.53, p=0.0005). This analysis confirms many recurrent LGD genes and CNV deletion syndromes (e.g., KANSL1, PAFAH1B1, RA1, etc.), consistent with a model of haploinsufficiency. We also identify genes with an excess of missense DNMs overlapping deletion syndromes (e.g., KIF1A and the 2q37 deletion) as well as duplication syndromes, such as recurrent MAPK3 missense mutations within the chromosome 16p11.2 duplication, recurrent CHD4 missense DNMs in the 12p13 duplication region, and recurrent WDFY4 missense DNMs in the 10q11.23 duplication region. Finally, we also identify pathogenic CNVs overlapping more than one recurrently mutated gene (e.g., Sotos and Kleefstra syndromes) raising the possibility that multiple gene-dosage imbalances may contribute to phenotypic complexity of these disorders. Network analyses of genes showing an excess of DNMs confirm previous well-known enrichments but also highlight new functional networks, including cell-specific enrichments in the D1+ and D2+ spiny neurons of the striatum for both recurrently mutated genes and genes where missense mutations cluster.

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Proteomic Analysis of Post-synaptic Density Fractions from Shank3 Mutant Mice Reveals Brain Region Specific Changes Relevant to Autism Spectrum Disorder

Cited by 68 publications

References 54 publications

Behavioral Phenotyping of an Improved Mouse Model of Phelan–McDermid Syndrome with a Complete Deletion of theShank3Gene

Behavioral Phenotyping of an Improved Mouse Model of Phelan–McDermid Syndrome with a Complete Deletion of theShank3Gene

Integrative Brain Transcriptome Analysis Reveals Region-Specific and Broad Molecular Changes in Shank3-Overexpressing Mice

Neurodevelopmental disease genes implicated byde novomutation and CNV morbidity

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