小鼠发育过程中七氟醚诱导的海马细胞类型和性别特异性效应的单核体图谱

Single-nucleus Atlas of Sevoflurane-induced Hippocampal Cell Type- and Sex-specific Effects during Development in Mice.

机构信息

Department of Anesthesiology, First Affiliated Hospital of Soochow University, Suzhou, China; Institute of Anesthesiology, Soochow University, Suzhou, China; Department of Pain Medicine, Dushu Lake Hospital Affiliated of Soochow University, Suzhou, China.

Department of Anesthesiology, First Affiliated Hospital of Soochow University, Suzhou, China; Institute of Anesthesiology, Soochow University, Suzhou, China.

出版信息

Anesthesiology. 2023 May 1;138(5):477-495. doi: 10.1097/ALN.0000000000004522.

DOI:10.1097/ALN.0000000000004522

PMID:36752736

Abstract

BACKGROUND

Multiple neonatal exposures to sevoflurane induce neurocognitive dysfunctions in rodents. The lack of cell type-specific information after sevoflurane exposure limits the mechanistic understanding of these effects. In this study, the authors tested the hypothesis that sevoflurane exposures alter the atlas of hippocampal cell clusters and have neuronal and nonneuronal cell type-specific effects in mice of both sexes.

METHODS

Neonatal mice were exposed to 3% sevoflurane for 2 h at postnatal days 6, 8, and 10 and analyzed for the exposure effects at postnatal day 37. Single-nucleus RNA sequencing was performed in the hippocampus followed by in situ hybridization to validate the results of RNA sequencing. The Morris Water Maze test was performed to test neurocognitive function.

RESULTS

The authors found sex-specific distribution of hippocampal cell types in control mice alongside cell type- and sex-specific effects of sevoflurane exposure on distinct hippocampal cell populations. There were important changes in male but not in female mice after sevoflurane exposure regarding the proportions of cornu ammonis 1 neurons (control vs. sevoflurane, males: 79.9% vs. 32.3%; females: 27.3% vs. 24.3%), dentate gyrus (males: 4.2% vs. 23.4%; females: 36.2% vs. 35.8%), and oligodendrocytes (males: 0.6% vs. 6.9%; females: 5.9% vs. 7.8%). In male but not in female mice, sevoflurane altered the number of significantly enriched ligand-receptor pairs in the cornu ammonis 1, cornu ammonis 3, and dente gyrus trisynaptic circuit (control vs. sevoflurane, cornu ammonis 1-cornu ammonis 3: 18 vs. 42 in males and 15 vs. 21 in females; cornu ammonis 1-dentate gyrus: 21 vs. 35 in males and 12 vs. 20 in females; cornu ammonis 3-dentate gyrus: 25 vs. 45 in males and 17 vs. 20 in females), interfered with dentate gyrus granule cell neurogenesis, hampered microglia differentiation, and decreased cornu ammonis 1 pyramidal cell diversity. Oligodendrocyte differentiation was specifically altered in females with increased expressions of Mbp and Mag. In situ hybridization validated the increased expression of common differentially expressed genes.

CONCLUSIONS

This single-nucleus RNA sequencing study reveals the hippocampal atlas of mice, providing a comprehensive resource for the neuronal and nonneuronal cell type- and sex-specific effects of sevoflurane during development.

摘要

背景

多次接触七氟醚会导致啮齿动物的神经认知功能障碍。七氟醚暴露后缺乏细胞类型特异性信息，限制了对这些影响的机制理解。在这项研究中，作者测试了以下假设：七氟醚暴露会改变海马体细胞簇图谱，并对雌雄小鼠的神经元和非神经元细胞类型产生特异性影响。

方法

新生小鼠在出生后第 6、8 和 10 天接受 3%七氟醚 2 小时的暴露，然后在出生后第 37 天分析暴露的影响。在海马体中进行单细胞 RNA 测序，然后进行原位杂交以验证 RNA 测序的结果。进行 Morris 水迷宫测试以测试神经认知功能。

结果

作者在对照组小鼠中发现了海马体细胞类型的性别特异性分布，同时发现了七氟醚暴露对不同海马体细胞群的细胞类型和性别特异性影响。七氟醚暴露后，雄性小鼠而非雌性小鼠的齿状回（雄性：4.2% vs. 23.4%；雌性：36.2% vs. 35.8%）和少突胶质细胞（雄性：0.6% vs. 6.9%；雌性：5.9% vs. 7.8%）的 Cornu Ammonis 1 神经元比例发生了重要变化。在雄性小鼠中，但在雌性小鼠中，七氟醚改变了 Cornu Ammonis 1、Cornu Ammonis 3 和 Dentate Gyrus 三突触回路中显著富集的配体-受体对的数量（雄性：Cornu Ammonis 1-Cornu Ammonis 3：18 对 42；雌性：15 对 21；Cornu Ammonis 1-Dentate Gyrus：21 对 35；雌性：12 对 20；Cornu Ammonis 3-Dentate Gyrus：25 对 45；雌性：17 对 20），干扰了齿状回颗粒细胞神经发生，阻碍了小胶质细胞分化，并减少了 Cornu Ammonis 1 锥体神经元的多样性。少突胶质细胞分化在雌性小鼠中特异性改变，Mbp 和 Mag 的表达增加。原位杂交验证了常见差异表达基因的表达增加。