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

Jasmin Morandell,Lisa S Knaus,Christoph P Dotter,Caroline Kreuzinger,Zoe Dobler,Georgi Dimchev,Johann G Danzl,Gaia Novarino,Bernadette Basilico,Armel Nicolas,Florian K M Schur,Lena A Schwarz,Saren Tasciyan,Christoph Sommer,Emanuele Cacci

doi:10.1038/s41467-021-23123-x

Jasmin Morandell, Lisa S Knaus + Show 13 more

Open Access

https://doi.org/10.1038/s41467-021-23123-x

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Abstract

De novo loss of function mutations in the ubiquitin ligase-encoding gene Cullin3 (CUL3) lead to autism spectrum disorder (ASD). In mouse, constitutive Cul3 haploinsufficiency leads to motor coordination deficits as well as ASD-relevant social and cognitive impairments. However, induction of Cul3 haploinsufficiency later in life does not lead to ASD-relevant behaviors, pointing to an important role of Cul3 during a critical developmental window. Here we show that Cul3 is essential to regulate neuronal migration and, therefore, constitutive Cul3 heterozygous mutant mice display cortical lamination abnormalities. At the molecular level, we found that Cul3 controls neuronal migration by tightly regulating the amount of Plastin3 (Pls3), a previously unrecognized player of neural migration. Furthermore, we found that Pls3 cell-autonomously regulates cell migration by regulating actin cytoskeleton organization, and its levels are inversely proportional to neural migration speed. Finally, we provide evidence that cellular phenotypes associated with autism-linked gene haploinsufficiency can be rescued by transcriptional activation of the intact allele in vitro, offering a proof of concept for a potential therapeutic approach for ASDs.

Highlights

De novo loss of function mutations in the ubiquitin ligase-encoding gene Cullin[3] (CUL3) lead to autism spectrum disorder (ASD)
Adult Cul3+/− mice present with hind limb clasping (Fig. 1a) and mild gait abnormalities, such as increased sway and stance length (Fig. 1b, c and Supplementary Fig. 2a), phenotypes which are observed in other ASD mouse models[19,20] and indicative of cerebellar dysfunctions[21]
Cul3+/− mice underperform when challenged on the accelerating RotaRod (Fig. 1d, d′), a task requiring formation and consolidation of a repetitive motor routine[22,23]

Summary

Introduction

De novo loss of function mutations in the ubiquitin ligase-encoding gene Cullin[3] (CUL3) lead to autism spectrum disorder (ASD). We found that Cul[3] controls neuronal migration by tightly regulating the amount of Plastin[3] (Pls3), a previously unrecognized player of neural migration. CUL3 ASDassociated genetic variants are most often de novo missense or loss of function (loF) mutations, dispersed throughout the entire gene and affecting distinct protein domains. Our analysis highlights a pivotal role for Cul[3] in brain development, identifies a new player of neuronal migration, and provides a proof of concept of CRISPR-mediated rescue of an ASD-linked genetic defect. Motor defects of Cul3+/− mice, do not affect exploratory behavior in the open field (Supplementary Fig. 2c), nor on the elevated plus maze, where Cul3+/− animals do not show differences in anxiety-like behaviors (Supplementary Fig. 2d)

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