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Repeated Ketamine Anesthesia during the Neonatal Period Impairs Hippocampal Neurogenesis and Long-Term Neurocognitive Function by Inhibiting Mfn2-Mediated Mitochondrial Fusion in Neural Stem Cells

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Abstract

The mechanism of ketamine-induced neurotoxicity development remains elusive. Mitochondrial fusion/fission dynamics play a critical role in regulating neurogenesis. Therefore, this study was aimed to evaluate whether mitochondrial dynamics were involved in ketamine-induced impairment of neurogenesis in neonatal rats and long-term synaptic plasticity dysfunction. In the in vivo study, postnatal day 7 (PND-7) rats received intraperitoneal (i.p.) injection of 40 mg/kg ketamine for four consecutive times at 1 h intervals. The present findings revealed that ketamine induced mitochondrial fusion dysfunction in hippocampal neural stem cells (NSCs) by downregulating Mitofusin 2 (Mfn2) expression. In the in vitro study, ketamine treatment at 100 μM for 6 h significantly decreased the Mfn2 expression, and increased ROS generation, decreased mitochondrial membrane potential and ATP levels in cultured hippocampal NSCs. For the interventional study, lentivirus (LV) overexpressing Mfn2 (LV-Mfn2) or control LV vehicle was microinjected into the hippocampal dentate gyrus (DG) 4 days before ketamine administration. Targeted Mfn2 overexpression in the DG region could restore mitochondrial fusion in NSCs and reverse the inhibitory effect of ketamine on NSC proliferation and its faciliatory effect on neuronal differentiation. In addition, synaptic plasticity was evaluated by transmission electron microscopy, Golgi-Cox staining and long-term potentiation (LTP) recordings at 24 h after the end of the behavioral test. Preconditioning with LV-Mfn2 improved long-term cognitive dysfunction after repeated neonatal ketamine exposure by reversing the inhibitory effect of ketamine on synaptic plasticity in the hippocampal DG. The present findings demonstrated that Mfn2-mediated mitochondrial fusion dysfunction plays a critical role in the impairment of long-term neurocognitive function and synaptic plasticity caused by repeated neonatal ketamine exposure by interfering with hippocampal neurogenesis. Thus, Mfn2 might be a novel therapeutic target for the prevention of the developmental neurotoxicity of ketamine.

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Data Availability

For further inquiries on the original data of this study, the corresponding authors can be contacted directly.

Abbreviations

BGS:

Brain growth spurt

BrdU:

Bromodeoxyuridine

CA:

Cornu ammonis

DCX:

Doublecortin

DG:

Dentate gyrus

Drp1:

Fission-related protein 1

fEPSP:

Field excitatory postsynaptic potential

GCL:

Granular cell layer

LTP:

Long-term potentiation

Mfn:

Mitofusins

MWM:

Morris water maze

NMDA:

N-methyl-D-aspartate

NSCs:

Neural stem cells

OPA1:

Optic atrophy 1

PND:

Postnatal day

PSD:

Postsynaptic density

ROS:

Reactive oxygen species

SD:

Sprague‒Dawley

TBS:

Theta burst stimulation

TEM:

Transmission Electron Microscopy

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Funding

This work is supported by the National Natural Science Foundation of China (grant number: 82171191, 81971051 to YW, 81901100 to HH) and Jiangsu Province Special Program for Young Medical Talent (QNRC2016587 to HH).

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Author notes

  1. He Huang and Ning Wang contributed equally to this work.

Authors and Affiliations

  1. Jiangsu Province Key Laboratory of Anesthesiology, NMPA Key Laboratory for Research and Evaluation of Narcotic and Psychotropic Drugs, Xuzhou Medical University, Xuzhou, 221004, Jiangsu, China

    He Huang, Ning Wang, Jia-Tao Lin, Yong-Kang Qiu, Wei-Feng Wu, Qiang Liu, Chen Chen, Hai-Bi Wang, Yan-Ping Liu, Wei Dong, Jie Wan & Yu-Qing Wu

  2. Department of Anesthesiology and Perioperative Medicine, The First Affliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, China

    He Huang

  3. Department of Anesthesiology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100021, China

    Hui Zheng

  4. Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, 221004, Jiangsu, China

    Cheng-Hua Zhou

Authors
  1. He Huang
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Contributions

Project conception: HH, YW, CZ, HZ

Study design: HH, YW, CZ, HZ

Performance of experiments: HH, NW, JL, YQ, WW, QL, CC,

Initial data collection and analysis: HW, YL, WD, JW

Final data analysis: HW, YL, WD, JW

Writing of paper: HH, YW, CZ, HZ

Corresponding authors

Correspondence to Hui Zheng, Cheng-Hua Zhou or Yu-Qing Wu.

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Ethics Approval

The experimental protocol was approved by the Institutional Animal Care and Ethics Committee of Xuzhou Medical University and in accordance with the Guide for the Care and Use of Laboratory Animals of the National Research Council.

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Huang, H., Wang, N., Lin, JT. et al. Repeated Ketamine Anesthesia during the Neonatal Period Impairs Hippocampal Neurogenesis and Long-Term Neurocognitive Function by Inhibiting Mfn2-Mediated Mitochondrial Fusion in Neural Stem Cells. Mol Neurobiol (2024). https://doi.org/10.1007/s12035-024-03921-2

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