Go to JCI Insight
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Publication alerts by email
  • Advertising
  • Job board
  • Contact
  • Clinical Research and Public Health
  • Current issue
  • Past issues
  • By specialty
    • COVID-19
    • Cardiology
    • Gastroenterology
    • Immunology
    • Metabolism
    • Nephrology
    • Neuroscience
    • Oncology
    • Pulmonology
    • Vascular biology
    • All ...
  • Videos
    • Conversations with Giants in Medicine
    • Video Abstracts
  • Reviews
    • View all reviews ...
    • Pancreatic Cancer (Jul 2025)
    • Complement Biology and Therapeutics (May 2025)
    • Evolving insights into MASLD and MASH pathogenesis and treatment (Apr 2025)
    • Microbiome in Health and Disease (Feb 2025)
    • Substance Use Disorders (Oct 2024)
    • Clonal Hematopoiesis (Oct 2024)
    • Sex Differences in Medicine (Sep 2024)
    • View all review series ...
  • Viewpoint
  • Collections
    • In-Press Preview
    • Clinical Research and Public Health
    • Research Letters
    • Letters to the Editor
    • Editorials
    • Commentaries
    • Editor's notes
    • Reviews
    • Viewpoints
    • 100th anniversary
    • Top read articles

  • Current issue
  • Past issues
  • Specialties
  • Reviews
  • Review series
  • Conversations with Giants in Medicine
  • Video Abstracts
  • In-Press Preview
  • Clinical Research and Public Health
  • Research Letters
  • Letters to the Editor
  • Editorials
  • Commentaries
  • Editor's notes
  • Reviews
  • Viewpoints
  • 100th anniversary
  • Top read articles
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Publication alerts by email
  • Advertising
  • Job board
  • Contact
Multimodal single-cell analyses reveal molecular markers of neuronal senescence in human drug-resistant epilepsy
Qianqian Ge, … , Li Shen, Jiadong Chen
Qianqian Ge, … , Li Shen, Jiadong Chen
Published March 3, 2025
Citation Information: J Clin Invest. 2025;135(5):e188942. https://doi.org/10.1172/JCI188942.
View: Text | PDF
Research Article Aging Neuroscience Article has an altmetric score of 9

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

  • Text
  • PDF
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 3

Multiplex EASI-FISH reveals molecularly defined PY1 and PY2 neurons as well as their electrophysiological and morphological characteristics.

Options: View larger image (or click on image) Download as PowerPoint
Multiplex EASI-FISH reveals molecularly defined PY1 and PY2 neurons as w...
(A) Schematic diagram illustrating the experimental procedures for loading the morphology of recorded neurons and subsequent staining with multiplex EASI-FISH. (B) Representative FISH images showing the expression of 6 differentially expressed genes (DEGs) NFKBIA, CDKN1A, CCL2, CUX2, NEFM, and NEFH in a ROI with biocytin loaded PY2 neuron in a brain slice from focal cortical dysplasia. Scale bar: 50 μm. Arrows and arrow heads indicate biocytin-loaded or nonbiocytin-loaded PY2 neurons, respectively. Outlined areas indicate the soma area of individual neurons. (C) Heatmap of 6 DEGs and Uniform Manifold Approximation and Projection (UMAP) analysis (D) of molecularly defined PY1 (217 neurons) and PY2 clusters (122 neurons) in an example brain slice. (E) Scatter plots showing the gene expression density of 6 DEGs in PY1 and PY2 neuron clusters. Red dashed line, PY1 cluster density fitting line; Blue dashed line, PY2 cluster density fitting line. (F–H) Representative images showing the expression of candidate marker genes in biocytin-labeled PY2 neuron (F), molecularly defined PY1 (G) and PY2 (H) pyramidal neurons. Outlined areas indicate the soma area of individual neurons. (I) Statistical results showing the cell size (left) and gene expression density (Slope of density fitting line) (right) in PY1 and PY2 neurons (n = 7,688 neurons, 7 brain slices from 4 individual samples). Volume, P = 0.0020; NFKBIA, P = 0.0319; CDKNIA, P = 0.0469; CCL2, P = 0.0974; CUX2, P = 0.5781; NEFM, P = 0.0361; NEFH, P = 0.0394. Each data point indicates the mean value from a sample; gray lines connect data points from the same sample. *P < 0.05; **P < 0.01. Normality and lognormality tests were used to determine whether the data fit the normal distribution, 2-tailed paired t test was employed when the data follows a normal distribution, whereas the nonparametric 2-tailed Wilcoxon signed rank test was utilized in cases where the data did not conform to a normal distribution.

Copyright © 2025 American Society for Clinical Investigation
ISSN: 0021-9738 (print), 1558-8238 (online)

Sign up for email alerts

Blogged by 1
Posted by 4 X users
On 1 Facebook pages
Referenced by 1 Bluesky users
6 readers on Mendeley
See more details