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Patient Mutations of the Intellectual Disability Gene KDM5C Downregulate Netrin G2 and Suppress Neurite Growth in Neuro2a Cells

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Abstract

The X-linked lysine (K)-specific demethylase 5C (KDM5C) gene plays an important role in brain development and behavior. It encodes a histone demethylase that is involved in gene regulation in neuronal differentiation and morphogenesis. When mutated, it causes neuropsychiatric symptoms, such as intellectual disability, delayed language development, epilepsy, and impulsivity. To better understand how the patient mutations affect neuronal development, we expressed KDM5C mutants in Neuro2a cells, a mouse neuroblastoma cell line. Retinoic acid (RA)-induced neurite growth was suppressed by the mutation KDM5C Y751C, KDM5C H514A, and KDM5C F642L, but not KDM5C D87G or KDM5C A388P. RNA-seq analysis indicated an upregulation of genes important for neuronal development, such as Ntng2, Enah, Gas1, Slit2, and Dscam, in response to the RA treatment in control Neuro2a cells transfected with GFP or wild-type KDM5C. In contrast, in cells transfected with KDM5C Y751C, these genes were not upregulated by RA. Ntng2 was downregulated in cells with KDM5C mutations, concordant with the lower levels of H3K4 methylation at its promoter. Moreover, knocking down Ntng2 in control Neuro2a cells led to the phenotype of short neurites similar to that of cells with KDM5C Y751C, whereas Ntng2 overexpression in the mutant cells rescued the morphological phenotype. These findings provide new insight into the pathogenesis of phenotypes associated with KDM5C mutations.

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Acknowledgments

We thank Dr. Shigeyoshi Itohara for kindly providing us with the Ntng2 plasmid. This study was supported by NIH grants MH096066 (JX), NS089896 (SI), GM113943 (CMD), and GM046883 (CMD), as well as an award from the Farrehi Family Foundation (SI).

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Correspondence to Jun Xu.

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All procedures involving animals were approved by the IACUC of the Washington State University (Pullman, WA), in accordance with the NIH Guide for the Care and Use of Laboratory Animals.

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Wei, G., Deng, X., Agarwal, S. et al. Patient Mutations of the Intellectual Disability Gene KDM5C Downregulate Netrin G2 and Suppress Neurite Growth in Neuro2a Cells. J Mol Neurosci 60, 33–45 (2016). https://doi.org/10.1007/s12031-016-0770-3

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