Multimodal single-cell analyses reveal molecular markers of neuronal senescence in human drug-resistant epilepsy
Qianqian Ge, Jiachao Yang, Fei Huang, Xinyue Dai, Chao Chen, Jingxin Guo, Mi Wang, Mengyue Zhu, Yijie Shao, Yuxian Xia, Yu Zhou, Jieqiao Peng, Suixin Deng, Jiachen Shi, Yiqi Hu, Huiying Zhang, Yi Wang, Xiaoqun Wang, Xiao-Ming Li, Zhong Chen, Yousheng Shu, Jun-Ming Zhu, Jianmin Zhang, Ying Shen, Shumin Duan, Shengjin Xu, Li Shen, Jiadong Chen
Qianqian Ge, Jiachao Yang, Fei Huang, Xinyue Dai, Chao Chen, Jingxin Guo, Mi Wang, Mengyue Zhu, Yijie Shao, Yuxian Xia, Yu Zhou, Jieqiao Peng, Suixin Deng, Jiachen Shi, Yiqi Hu, Huiying Zhang, Yi Wang, Xiaoqun Wang, Xiao-Ming Li, Zhong Chen, Yousheng Shu, Jun-Ming Zhu, Jianmin Zhang, Ying Shen, Shumin Duan, Shengjin Xu, Li Shen, Jiadong Chen
View: Text | PDF
Research Article Aging Neuroscience

Multimodal single-cell analyses reveal molecular markers of neuronal senescence in human drug-resistant epilepsy

Abstract

The histopathological neurons in the brain tissue of drug-resistant epilepsy exhibit aberrant cytoarchitecture and imbalanced synaptic circuit function. However, the gene expression changes of these neurons remain unknown, making it difficult to determine the diagnosis or to dissect the mechanism of drug-resistant epilepsy. By integrating whole-cell patch clamp recording and single-cell RNA-seq approaches, we identified a transcriptionally distinct subset of cortical pyramidal neurons. These neurons highly expressed genes CDKN1A (P21), CCL2, and NFKBIA, which associate with mTOR pathway, inflammatory response, and cellular senescence. We confirmed the expression of senescent marker genes in a subpopulation of cortical pyramidal neurons with enlarged soma size in the brain tissue of drug-resistant epilepsy. We further revealed the expression of senescent cell markers P21, P53, COX2, γ-H2AX, and β-Gal, and reduction of nuclear integrity marker Lamin B1 in histopathological neurons in the brain tissue of patients with drug-resistant epilepsy with different pathologies, but not in control brain tissue with no history of epilepsy. Additionally, chronic, but not acute, epileptic seizures induced senescent marker expression in cortical neurons in mouse models of drug-resistant epilepsy. These results provide important molecular markers for histopathological neurons and what we believe to be new insights into the pathophysiological mechanisms of drug-resistant epilepsy.

Authors

Qianqian Ge, Jiachao Yang, Fei Huang, Xinyue Dai, Chao Chen, Jingxin Guo, Mi Wang, Mengyue Zhu, Yijie Shao, Yuxian Xia, Yu Zhou, Jieqiao Peng, Suixin Deng, Jiachen Shi, Yiqi Hu, Huiying Zhang, Yi Wang, Xiaoqun Wang, Xiao-Ming Li, Zhong Chen, Yousheng Shu, Jun-Ming Zhu, Jianmin Zhang, Ying Shen, Shumin Duan, Shengjin Xu, Li Shen, Jiadong Chen

×

Figure 7

Neuronal senescent markers are induced by epileptic activities in mouse model of drug-resistant epilepsy.

Options: View larger image (or click on image) Download as PowerPoint
Neuronal senescent markers are induced by epileptic activities in mouse ...
(A) Schematic diagram showing the experimental design of video EEG recording and rapamycin injection (i.p.) of CamKII:Pten–/– mice. (B) Representative traces showing generalized seizures (GS) from EEG recordings of vehicle-treated (black traces, n = 3 mice) and rapamycin-treated (red traces, n = 3 mice) CamKII:Pten–/– mice. (C) Average number and duration of GS were substantially reduced in rapamycin-treated compared with vehicle-treated CamKII:Pten–/– mice (n = 6 mice per group). ****P < 0.0001. 2-tailed t test. (D) Rapamycin treatment extended the lifespan of CamKII:Pten–/– mice. Black trace: vehicle-treated group (n = 18 mice); red trace: rapamycin-treated group (n = 10 mice). P = 0.0047, Log-rank (Mantel-Cox) test. (E and F) Representative images showing the expression of senescent markers (SA-β-Gal, COX2, P21, P53, γ-H2AX) and reduction of nuclear integrity marker Lamin B1 in CamKII:Pten–/– mice with status epilepticus, but not in CamKII:Pten–/– mice that had not developed generalized seizures at 5 postnatal weeks (5 wk), or in control Ptenfl/fl mice at 12 wk. Rapamycin treatment repressed senescent marker expression in CamKII:Pten–/– mice. Expression of Lamin B1 can be restored after rapamycin treatment of CamKII:Pten–/– mice. Arrows indicate coexpression of senescent markers with NeuN. Scale bar, 50 μm; 10 μm for zoom-in images. (G) Statistical results showing the proportion of senescent markers and NeuN coexpressing neurons in total cortical neurons. n = 3 mice per group. Data are presented as mean ± SEM. Kruskal-Wallis test with Dunn’s multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.