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Differential epitranscriptome and proteome modulation in the brain of neonatal mice exposed to isoflurane or sevoflurane

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Abstract

Background

Repeated neonatal exposure to anesthetics may disturb neurodevelopment and cause neuropsychological disorders. The m6A modification participates in the gene regulation of neurodevelopment in mouse fetuses exposed to anesthetics. This study aims to explore the underlying molecular mechanisms of neurotoxicity after early-life anesthesia exposure.

Methods

Mice were exposed to isoflurane (1.5%) or sevoflurane (2.3%) for 2 h daily during postnatal days (PND) 7–9. Sociability, spatial working memory, and anxiety-like behavior were assessed on PND 30–35. Synaptogenesis, epitranscriptome m6A, and the proteome of brain regions were evaluated on PND 21.

Results

Both isoflurane and sevoflurane produced abnormal social behaviors at the juvenile age, with different sociality patterns in each group. Synaptogenesis in the hippocampal area CA3 was increased in the sevoflurane-exposed mice. Both anesthetics led to numerous persistent m6A-induced alterations in the brain, associated with critical metabolic, developmental, and immune functions. The proteins altered by isoflurane exposure were mainly associated with epilepsy, ataxia, and brain development. As for sevoflurane, the altered proteins were involved in social behavior.

Conclusions

Social interaction, the modulation patterns of the m6A modification, and protein expression were altered in an isoflurane or sevoflurane-specific way. Possible molecular pathways involved in brain impairment were revealed, as well as the mechanism underlying behavioral deficits following repeated exposure to anesthetics in newborns.

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Funding

This work was supported by the National Natural Science Foundation of China (81971060, 81401279) and the Natural Science Foundation of Hubei Province of China (2020CFB342).

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Author notes

  1. Yanqiong Wu and Zeyong Yang contributed equally to this work.

Authors and Affiliations

  1. Institute of Anesthesiology & Pain (IAP), Hubei Clinical Research Center for Precise Diagnosis and Treatment of Liver Cancer, Department of Anesthesiology, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, China

    Yanqiong Wu, Shanchun Su, Xueqin Xu, Yang Li, Xiaohui Li, Yan Gao, Dongsheng Sun, Shengjun Wan, Mingjin Pen & Changbin Ke

  2. Department of Anesthesiology, International Peace Maternity and Child Health Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, 200030, China

    Zeyong Yang

  3. Department of Instrument Science and Engineering, Key Lab. for Thin Film and Microfabrication Technology of Ministry of Education, School of Electronic Information and Electronic Engineering, Institute of Nano Biomedicine and Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China

    Weiling Jin

  4. Institute of Cancer Neuroscience, Medical Frontier Innovation Research Center, The First Hospital of Lanzhou University, The First Clinical Medical College of Lanzhou University, Lanzhou, 730000, People’s Republic of China

    Weiling Jin

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  1. Yanqiong Wu
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Contributions

CK, WJ conceived and designed the experiments. YW conducted the study and wrote the manuscript. CS, SW, and MP helped conduct the study and collect the data. XX, YL, XL, and DS helped conduct the study. ZY helped to analyze the data and revise the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Weiling Jin or Changbin Ke.

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Ethical approval was obtained from the medical/animal ethical committee of the HuBei University of Medicine (China). Informed consent was obtained where applicable.

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All authors have read and agreed to the published version of the manuscript.

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The authors declare no competing interests.

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Keywords

• Neonatal exposure to isoflurane or sevoflurane caused sociability deficits at the juvenile age.

• Neonatal exposure to isoflurane or sevoflurane induced m6A and proteome modification.

• Neonatal exposure to sevoflurane disturbed synaptogenesis.

Supplementary Information

Below is the link to the electronic supplementary material.

10565_2022_9701_MOESM1_ESM.jpg

Supplementary file1 The distribution of m6A peak in the genome and the remarkablely alteredm6A peaks after repeated neonatal exposure to isoflurane or sevoflurane. (A) Thepercentage of m6A peaks identified in the genome regions. (B and C) Heatmapsshowing the enrichment of m6A peak (red, log2FC > 1; blue, log2FC < -1) and itsannotated genes in the isoflurane (B) and sevoflurane (C) groups. FC, fold change. (JPG 2323 KB)

10565_2022_9701_MOESM2_ESM.jpg

Supplementary file2 m6A installation protein expression in the brainThe expression of m6A installation protein in the cortex were examined using westernblot of normal, isoflurane and sevoflurane exposure mice. Quantitative analysis ofbands showed on the right (compare to normal, * p<0.05). (JPG 2059 KB)

10565_2022_9701_MOESM3_ESM.jpg

Supplementary file3 Protein expression in the cerebral cortex and hippocampus after repeatedneonatal exposure to isoflurane or sevoflurane. (A and A’) Volcano plot (A) andheatmap (A’) of the differentially expressed proteins in the cerebral cortex betweenisoflurane and control. (B and B’) Volcano plot (B) and heatmap (B’) of thedifferentially expressed proteins in the cerebral cortex between sevoflurane and control.(C and C’) Volcano plot (C) and heatmap (C’) of the differentially expressed proteinsin the hippocampus between isoflurane and control. (D and D’) Volcano plot (D) andheatmap (D’) of the differentially expressed proteins in the hippocampus betweensevoflurane and control. Red dots in volcano plot represent the significantly increasedproteins (fold change > 1.5, p < 0.05), blue dots in volcano plot represent thesignificantly decreased proteins (fold change < 0.67, p < 0.05). (JPG 3902 KB)

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Wu, Y., Yang, Z., Su, S. et al. Differential epitranscriptome and proteome modulation in the brain of neonatal mice exposed to isoflurane or sevoflurane. Cell Biol Toxicol 39, 2133–2148 (2023). https://doi.org/10.1007/s10565-022-09701-9

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