Mutations of CNTNAP1 led to defects in neuronal development
Wanxing Li, Lin Yang, Chuanqing Tang, Kaiyi Liu, Yulan Lu, Huijun Wang, Kai Yan, Zilong Qiu, Wenhao Zhou
Wanxing Li, Lin Yang, Chuanqing Tang, Kaiyi Liu, Yulan Lu, Huijun Wang, Kai Yan, Zilong Qiu, Wenhao Zhou
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Research Article Genetics Neuroscience

Mutations of CNTNAP1 led to defects in neuronal development

Abstract

Mutations of CNTNAP1 were associated with myelination disorders, suggesting the role of CNTNAP1 in myelination processes. Whether CNTNAP1 may have a role in early cortical neuronal development is largely unknown. In this study, we identified 4 compound heterozygous mutations of CNTNAP1 in 2 Chinese families. Using mouse models, we found that CNTNAP1 is highly expressed in neurons and is located predominantly in MAP2+ neurons during the early developmental stage. Importantly, Cntnap1 deficiency results in aberrant dendritic growth and spine development in vitro and in vivo, and it delayed migration of cortical neurons during early development. Finally, we found that the number of parvalbumin+ neurons in the cortex and hippocampus of Cntnap1–/– mice is strikingly increased by P15, suggesting that excitation/inhibition balance is impaired. Together, this evidence elucidates a critical function of CNTNAP1 in cortical development, providing insights underlying molecular and circuit mechanisms of CNTNAP1-related disease.

Authors

Wanxing Li, Lin Yang, Chuanqing Tang, Kaiyi Liu, Yulan Lu, Huijun Wang, Kai Yan, Zilong Qiu, Wenhao Zhou

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Figure 2

D1140Y and G1251R expression levels in 293T cells and mouse primary cortical neurons.

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D1140Y and G1251R expression levels in 293T cells and mouse primary cort...
(A) Representative Western blots of 293T cells and mouse primary cortical neurons transfected with Flag-tagged CNTNAP1-WT, CNTNAP1-D1140Y, and CNTNAP1-G1251R. β-Actin was used as a loading control. (B) Relative protein expression of Flag-tagged CNTNAP1-WT, CNTNAP1-D1140Y, and CNTNAP1-G1251R in 293T cells and mouse primary cortical neurons (represents the ratio of Flag in the upper panel to actin in the lower panel). The band intensities were determined by ImageJ. Statistical significance was evaluated by ordinary 1-way ANOVA: **P = 0.0028; *P = 0.0413. Data are shown as mean ± SEM. All data were collected from 4 independent experiments. (C) Representative Western blots of 293T cells transfected with Flag-tagged CNTNAP1-WT, CNTNAP1-D1140Y, and CNTNAP1-G1251R and treated with cycloheximide (CHX; 20 μg/mL) for 0, 3, 6, and 12 hours. (D) Relative protein expression of Flag-tagged CNTNAP1-WT, CNTNAP1-D1140Y, and CNTNAP1-G1251R in 293T cells (represents the ratio of Flag in the upper panel to actin in the lower panel). The band intensities were determined by ImageJ. Statistical significance was evaluated by ordinary 1-way ANOVA. Data are shown as mean ± SEM. All data were collected from 3 independent experiments. (E) Relative RNA expression of Flag-tagged CNTNAP1-WT, CNTNAP1-D1140Y, and CNTNAP1-G1251R in mouse primary cortical neurons. The level was normalized to that of WT group. Statistical significance was evaluated by ordinary 1-way ANOVA. Data are shown as mean ± SEM. All data were collected from 3 independent experiments.