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Restoration of nNOS Expression Rescues Autistic-Like Phenotypes Through Normalization of AMPA Receptor-Mediated Neurotransmission

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Abstract

Autism spectrum disorder (ASD) is associated with a range of abnormalities characterized by deficits in socialization, communication, repetitive behaviors, and restricted interests. We have recently shown that neuronal nitric oxide synthase (nNOS) expression was decreased in the basolateral amygdala (BLA) of mice after postnatal valproic acid exposure. Neuronal activity-regulated pentraxin (Narp) could contribute to the regulation of the GluA4 2-amino-3-(5-methyl-3-oxo-1,2-oxazol-4-yl) propanoic acid (AMPA) subunits which are predominantly expressed in interneurons. However, the specific role of nNOS re-expression on excitatory neurotransmitter with relevance to ASD core symptoms in VPA-treated animals remains to be elucidated. Herein, nNOS overexpression using a lentiviral vector and L-arginine-activating PI3K-Akt-mTOR signaling can restore nNOS expression in the BLA induced by VPA. Restoration of nNOS expression in these mice was sufficient to reduce the severity of ASD-like behavioral patterns such that animals exhibited decreases in abnormal social interactions and communication, stereotyped/repetitive behaviors, and anxiety-like traits. Most strikingly, re-expression of nNOS upregulated surface expression of Narp and GluA4 in nNOS-positive interneuron as shown by immunoprecipitation and Western blotting. Whole-cell patch-clamp recordings demonstrated that restoration of nNOS had a significant enhancing effect on AMPA receptor-mediated excitatory glutamatergic synaptic neurotransmission, which was inhibited by disturbing the interaction between Narp and GluA4 in acutely dissociated BLA slices. Overall, these data offer a scientific basis for the additional study of nNOS re-expression as a promising therapeutic target by correcting AMPA receptor-mediated synaptic function in ASD and related neurodevelopmental disorders.

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Data Availability

All data generated for this study are available on request to the corresponding author.

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Funding

We gratefully acknowledge the sponsorship from the National Natural Science Foundation of China (81901387), the Scientific and Technological Project of Henan (232102311006, 212102310034, 232102310077), the Medical Science and Technique Program of Henan (LHGJ20210632), the Natural Science Foundation of Henan (232300421086), the Henan Neural Development Engineering Research Center for Children Foundation (SG202202), and the Joint Fund of Henan Provincial Science and Technology Research (225200810114).

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Authors and Affiliations

  1. Henan Children’s Hospital, Zhengzhou Children’s Hospital, Henan Key Laboratory of Children’s Genetics and Metabolic Diseases, Henan Children’s Neurodevelopment Engineering Research Center, Children’s Hospital Affiliated to Zhengzhou University, Zhengzhou, China

    Xiaona Wang, Yaodong Zhang, Shuying Luo, Ke Zhao, Yongtao Duan & Shunan Hu

  2. Department of Rehabilitation, Children’s Hospital Affiliated to Zhengzhou University, Zhengzhou, China

    Chao Gao

  3. Department of Neurology, Children’s Hospital Affiliated to Zhengzhou University, Zhengzhou, China

    Daoqi Mei

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  1. Xiaona Wang
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Contributions

XW designed the research and wrote the manuscript. KZ conducted the viral infusion surgeries. SL and SH performed the behavioral characterization and analyzed the data. YD, DM, and CG carried out western blotting. XW performed electrophysiological experiments and analyzed the data as well. XW and YZ supervised all aspects of the research. All authors have read and approved the final version of the manuscript.

Corresponding author

Correspondence to Xiaona Wang.

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The animal study was reviewed and approved by the Institutional Animal Ethics Committee of Zhengzhou University, following the NIH Guidelines for the Care and Use of Laboratory Animals.

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Wang, X., Zhang, Y., Luo, S. et al. Restoration of nNOS Expression Rescues Autistic-Like Phenotypes Through Normalization of AMPA Receptor-Mediated Neurotransmission. Mol Neurobiol (2024). https://doi.org/10.1007/s12035-024-03997-w

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