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KCTD13-mediated ubiquitination and degradation of GluN1 regulates excitatory synaptic transmission and seizure susceptibility

Cell Death & Differentiation volume 30, pages 1726–1741 (2023)Cite this article

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Abstract

Temporal lobe epilepsy (TLE) is the most common and severe form of epilepsy in adults; however, its underlying pathomechanisms remain elusive. Dysregulation of ubiquitination is increasingly recognized to contribute to the development and maintenance of epilepsy. Herein, we observed for the first time that potassium channel tetramerization domain containing 13 (KCTD13) protein, a substrate-specific adapter for cullin3-based E3 ubiquitin ligase, was markedly down-regulated in the brain tissue of patients with TLE. In a TLE mouse model, the protein expression of KCTD13 dynamically changed during epileptogenesis. Knockdown of KCTD13 in the mouse hippocampus significantly enhanced seizure susceptibility and severity, whereas overexpression of KCTD13 showed the opposite effect. Mechanistically, GluN1, an obligatory subunit of N-methyl-D-aspartic acid receptors (NMDARs), was identified as a potential substrate protein of KCTD13. Further investigation revealed that KCTD13 facilitates lysine-48-linked polyubiquitination of GluN1 and its degradation through the ubiquitin-proteasome pathway. Besides, the lysine residue 860 of GluN1 is the main ubiquitin site. Importantly, dysregulation of KCTD13 affected membrane expression of glutamate receptors and impaired glutamate synaptic transmission. Systemic administration of the NMDAR inhibitor memantine significantly rescued the epileptic phenotype aggravated by KCTD13 knockdown. In conclusion, our results demonstrated an unrecognized pathway of KCTD13-GluN1 in epilepsy, suggesting KCTD13 as a potential neuroprotective therapeutic target for epilepsy.

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Fig. 1: Expression and localization of KCTD13 in the brain tissues of mice and patients with TLE.
Fig. 2: KCTD13 regulates seizure susceptibility and severity.
Fig. 3: KCTD13 regulates glutamatergic synaptic transmission.
Fig. 4: KCTD13 interacts with GluN1 and negatively regulates GluN1 protein expression.
Fig. 5: KCTD13 regulates the ubiquitination of GluN1 and mediates its degradation via the proteasome pathway.
Fig. 6: KCTD13 promotes K48-linked polyubiquitination of GluN1 at lysine residue 860 and regulates the surface and synaptosomal expression of glutamate receptors.
Fig. 7: KCTD13 regulates the number of functional glutamatergic synapses.
Fig. 8: KCTD13 regulates NMDAR-mediated glutamatergic synaptic transmission.

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Data availability

Data supporting the conclusions in the paper are present in the paper and the Supplementary Materials. Additional data are available from the corresponding author.

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Acknowledgements

We express our gratitude to the patients for the donations of brain tissue and their time and effort devoted to the consent process.

Funding

This work was supported by National Natural Science Foundation of China (Nos. 81922023, 82271496, 81873788, 82001378 and 82171440), Natural Science Foundation of Chongqing (CSTB2022NSCQ-MSX0747 and cstc2021ycjh-bgzxm0035). Future Medical Youth Innovation Team Program of Chongqing Medical University (No. W0043), the Fifth Senior Medical Talents Program of Chongqing for Young and Middle-aged, Middle-aged Medical Excellence Team Program of Chongqing, and Chongqing chief expert studio project, China. Science and Technology Research Program of Chongqing Education Commission (KJQN202200435), Chongqing Talents: Exceptional Young Talents Project (CQYC202005014).

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Authors and Affiliations

  1. Department of Neurology, The First Affiliated Hospital of Chongqing Medical University, Chongqing Key Laboratory of Neurology, Chongqing, 400016, China

    Juan Gu, Pingyang Ke, Haokun Guo, Jing Liu, Yan Liu, Xin Tian, Xin Xu, Demei Xu, Yuanlin Ma, Xuefeng Wang & Fei Xiao

  2. Department of Neurology, The Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, Luzhou, 646000, China

    Juan Gu

  3. State Key Laboratory of Natural and Biomimetic Drugs, Department of Molecular and Cellular Pharmacology, School of Pharmaceutical Sciences, Peking University Health Science Center, 100191, Beijing, China

    Zhuo Huang

  4. Institute for Brain Science and Disease of Chongqing Medical University, Chongqing, 400016, China

    Fei Xiao

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Contributions

JG, FX, PK and XW designed the study; JG, PK, HG, JL, YL, XT, XX and DX performed the experiments; DX and YM participated in the data and sample collection; JG, PK, HG and ZH analyzed data; JG, FX and ZH wrote the manuscript; XW and FX oversaw the project. All authors read and approved the final paper.

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Correspondence to Xuefeng Wang or Fei Xiao.

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The authors declare no competing interests.

Ethical approval

All animal experiments were approved by the Ethics Committee of Chongqing Medical University. Human brain tissue samples from patients with TLE or TBI were collected from the First Affiliated Hospital of Chongqing Medical University. Written informed consent was obtained from patients for the use of brain tissue and access to medical records for research purposes. The collection and use of all specimens were approved by the Ethics Committee of The First Affiliated Hospital of Chongqing Medical University and in accordance with the Declaration of Helsinki.

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Gu, J., Ke, P., Guo, H. et al. KCTD13-mediated ubiquitination and degradation of GluN1 regulates excitatory synaptic transmission and seizure susceptibility. Cell Death Differ 30, 1726–1741 (2023). https://doi.org/10.1038/s41418-023-01174-5

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