Behavioral, circuitry, and molecular aberrations by region-specific deficiency of the high-risk autism gene Cul3

Rapanelli, Maximiliano; Tan, Tao; Wang, Wei; Wang, Xue; Wang, Zijun; Zhong, Ping; Frick, Luciana Romina; Qin, Luye; Ma, Kaijie; Qu, Jun; Yan, Zhen

doi:10.1038/s41380-019-0498-x

Cited by 52 publications

(87 citation statements)

References 47 publications

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“…5c and Supplementary Table 4). Overall, and in agreement with previous observations 30-32 , the observed fold changes were mild, in line with the hypothesis that the ubiquitylated isoform of a protein represents only a small fraction of the total pool of that given protein at any time point 33 . We noticed, however, that proteins with abnormal levels in Cul3 homozygous mutants are significantly enriched for high confidence ASD-risk genes ( Cul3 fl/fl Emx1-Cre at 20% FDR, 65 genes, p= 0.0329).…”

Section: Resultssupporting

confidence: 92%

“…Second, we found that the only protein consistently up regulated in mutant animals is Pls3, a still poorly characterized actin bundling protein 36,37 . Pls3 is altered in the developing and adult cortex, the adult hippocampus, but not in the adult striatum 31,32 . Interestingly, a pathogenic variant of PLS3 has recently also been described in a patient with idiopathic osteoporosis and ASD 38 .…”

Section: Resultsmentioning

confidence: 96%

“…However, when we compared our data-set with recently published proteomic analysis of conditional Cul3 knockouts at adult stages 31,32 , we made a few interesting observations. First, in all data-sets we found down-regulation of Map2, a microtubule associated protein and known component of the neuronal cytoskeleton.…”

Section: Resultsmentioning

confidence: 99%

“…Importantly, while two recent studies 31,32 analyzed the effect of Cul3 deficiency and haploinsufficiency in the adult mouse brain, both studies fell short of studying the developmental consequences of Cul3 mutations in detail and employed Cre-lines that may miss critical developmental issues. In contrast, here we focused on the constitutive effects of Cul3 haploinsufficiency and discovered its crucial role in cell migration and proper organization of the cortex.…”

Section: Discussionmentioning

confidence: 99%

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Cul3 regulates cytoskeleton protein homeostasis and cell migration during a critical window of brain development

Morandell

Schwarz

Basilico

et al. 2020

Preprint

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11De novo loss of function mutations in the ubiquitin ligase-encoding gene Cullin3 (CUL3) lead to 12 autism spectrum disorder (ASD). Here, we used Cul3 mouse models to evaluate the 13 consequences of Cul3 mutations in vivo. Our results show that Cul3 haploinsufficient mice 14 exhibit deficits in motor coordination as well as ASD-relevant social and cognitive impairments. 15Cul3 mutant brain displays cortical lamination abnormalities due to defective neuronal migration 16 and reduced numbers of excitatory and inhibitory neurons. In line with the observed abnormal 17 columnar organization, Cul3 haploinsufficiency is associated with decreased spontaneous 18 excitatory and inhibitory activity in the cortex. At the molecular level, employing a quantitative 19 proteomic approach, we show that Cul3 regulates cytoskeletal and adhesion protein abundance 20 in mouse embryos. Abnormal regulation of cytoskeletal proteins in Cul3 mutant neuronal cells 21 results in atypical organization of the actin mesh at the cell leading edge, likely causing the 22 observed migration deficits. In contrast to these important functions early in development, Cul3 23 deficiency appears less relevant at adult stages. In fact, induction of Cul3 haploinsufficiency in 24 adult mice does not result in the behavioral defects observed in constitutive Cul3 25

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Section: Resultssupporting

confidence: 92%

Section: Resultsmentioning

confidence: 96%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Cul3 regulates cytoskeleton protein homeostasis and cell migration during a critical window of brain development

Morandell

Schwarz

Basilico

et al. 2020

Preprint

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“…Thus, downregulation of either KCTD13 or Cul3 are expected to upregulate the levels of RhoA. A recent study of conditional Cul3 model also observed RhoA upregulation in the cortex 29 . Most importantly, RhoA is a key regulator of actin cytoskeleton remodeling, neurite outgrowth, migration and spine formation during early cortical development 30 , and we observed defects in many of these pathways in Cul3 +/mouse.…”

Section: Discussionmentioning

confidence: 93%

Autism-linked Cullin3 germline haploinsufficiency impacts cytoskeletal dynamics and cortical neurogenesis through RhoA signaling

Amar

Pramod

Herrera

et al. 2020

Preprint

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E3-ubiquitin ligase Cullin3 (Cul3) is a high confidence risk gene for Neurodevelopmental Disorders (NDD) including Autism Spectrum Disorder (ASD) and Developmental Delay (DD). We generated haploinsufficient Cul3 mouse model to investigate brain anatomy, behavior, molecular, cellular, and circuit-level mechanisms dysregulated by Cul3 mutations. Brain MRI of Cul3 mutant animals found profound abnormalities in half of brain regions, including decreased volume of cortical regions and increased volume of subcortical regions. Cul3 mutant mice exhibited social and cognitive deficits, and hyperactive behavior. Spatiotemporal transcriptomic and proteomic profiling of the brain implicated neurogenesis and cytoskeletal defects as key drivers of Cul3 functional impact. Cortical neurons from mutant mice had reduced dendritic length and loss of filamentous actin puncta, along with reduced spontaneous network activity. Inhibition of small GTPase RhoA, a molecular substrate of Cul3 ligase, rescued dendrite length phenotype. This study identified RhoA signaling as a potential mechanism, through which Cul3 mutation impacts early brain development.introduced a germline 1bp insertion in exon 6 of Cul3, immediately adjacent to G754T (E246X) mutation detected in an ASD patient (O'Roak et al., 2012) (Figure 1B). Extensive characterization of the mutant Cul3 +/mice at multiple levels including brain neuroanatomy, behavior, transcriptomic and proteomic profiling of several brain region at three developmental time points, provided a detailed picture of the functional impact of Cul3 mutation (Figure 1C). Our results demonstrate that Cul3 haploinsufficiency severely impacts the developing brain and leads to social deficits and hyperactivity in mice. Dysregulation of genes and proteins involved in actin, intermediate filament and binding, pointed to cytoskeletal defects. Reduced synaptic connectivity and dendritic length were among most notable neuronal phenotypes observed in Cul3 +/mice. Importantly, upregulation of one of the Cul3 substrates, small GTPase RhoA, a regulator of cytoskeleton, neuronal growth and migration, linked observed molecular defects with Cul3 pathology. Treatment of cortical neurons with RhoA inhibitor Rhosin rescued dendritic length phenotype. ResultsCul3 +/-mice have severe brain anatomical defectsWe introduced a 1bp frame-shifting insertion with CRISPR/Cas9 into exon-6 of Cul3 one base pair upstream from the nucleotide mutated in an ASD patient (O'Roak et al., 2012) to generate Cul3 +/haploinsufficient mouse model on C57BL/6N background (STAR Methods). A heterozygous Cul3 +/founder mouse harboring the insertion was expanded via breeding with wildtype (WT) C57BL/6N mice for at least four generations to eliminate CRISPR off-target effects. In agreement with previous observations (Singer et al., 1999), homozygous Cul3 −/− mutant animals were not viable. The heterozygous Cul3 +/mice were viable, reached a normal lifespan, and were fertile irrespective of sex. However, Cul3 +/mice had reduced body weight (P<0.001 for mal...

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Small Interfering RNAs and RNA Therapeutics in Cardiovascular Diseases

Bansal

Arora

2020

Advances in Experimental Medicine and Biology

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Behavioral, circuitry, and molecular aberrations by region-specific deficiency of the high-risk autism gene Cul3

Cited by 52 publications

References 47 publications

Cul3 regulates cytoskeleton protein homeostasis and cell migration during a critical window of brain development

Cul3 regulates cytoskeleton protein homeostasis and cell migration during a critical window of brain development

Autism-linked Cullin3 germline haploinsufficiency impacts cytoskeletal dynamics and cortical neurogenesis through RhoA signaling

Small Interfering RNAs and RNA Therapeutics in Cardiovascular Diseases

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