Hypothalamic neural communication characteristics in 3 mouse models based on sequencing analysis: Attenuated inhibitory regulation with dominant roles of glutamate, NO, and electrical synapse signaling

ZHOU Lianyu; FEI Xinghang; YIN Huachun; ZANG Zhenle; ZHENG Xin; FAN Xiaotang; LI Song; YANG Hui

doi:10.16016/j.2097-0927.202512099

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Hypothalamic neural communication characteristics in 3 mouse models based on sequencing analysis: Attenuated inhibitory regulation with dominant roles of glutamate, NO, and electrical synapse signaling

Neuroscience | 更新时间：2026-03-30

- Hypothalamic neural communication characteristics in 3 mouse models based on sequencing analysis: Attenuated inhibitory regulation with dominant roles of glutamate, NO, and electrical synapse signaling
- Journal of Army Medical University Vol. 48, Issue 6, Pages: 687-700(2026)
- 作者机构：
  
  1.陆军军医大学（第三军医大学），第二附属医院神经外科，重庆
  2.陆军军医大学（第三军医大学），心理系军事认知心理学教研室，重庆
- 作者简介：
  
  LI Song, E-mail：Tmmuls@tmmu.edu.cn
  YANG Hui, E-mail：Yanghui@tmmu.edu.cn
- 基金信息：
  
  the National Natural Science Foundation of China(32471027);the Special Project of Science and Technology Innovation and Application Development of Chongqing(CSTB2024TIAD-STX0044);the Natural Science Foundation of Chongqing(CSTB2024NSCQ-MSX0021)
- DOI：10.16016/j.2097-0927.202512099
  CLC： R589.2;R338.2;Q811.4
- Received：18 December 2025，
  
  Revised：2026-01-25，
  
  Published：30 March 2026
- 稿件说明：
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周连宇，费兴杭，尹华春，等.测序解析3种肥胖模型小鼠下丘脑神经通讯特征：抑制性调控减弱，谷氨酸、NO与电突触信号各显主导［J］.陆军军医大学学报，2026，48（06）：687-700.DOI：10.16016/j.2097-0927.202512099.

ZHOU L Y，FEI X H，YIN H C，et al.Hypothalamic neural communication characteristics in 3 mouse models based on sequencing analysis： Attenuated inhibitory regulation with dominant roles of glutamate， NO， and electrical synapse signaling［J］.J Army Med Univ，2026，48（06）：687-700.DOI：10.16016/j.2097-0927.202512099.
周连宇，费兴杭，尹华春，等.测序解析3种肥胖模型小鼠下丘脑神经通讯特征：抑制性调控减弱，谷氨酸、NO与电突触信号各显主导［J］.陆军军医大学学报，2026，48（06）：687-700.DOI：10.16016/j.2097-0927.202512099. DOI：

ZHOU L Y，FEI X H，YIN H C，et al.Hypothalamic neural communication characteristics in 3 mouse models based on sequencing analysis： Attenuated inhibitory regulation with dominant roles of glutamate， NO， and electrical synapse signaling［J］.J Army Med Univ，2026，48（06）：687-700.DOI：10.16016/j.2097-0927.202512099. DOI：

摘要

目的

对比高脂饮食（high-fat diet，HFD）、瘦素突变（ob/ob）、瘦素受体突变（db/db）3种不同肥胖模型小鼠下丘脑细胞构成以及神经通讯网络变化识别可跨病因验证的普适机制线索、寻找适用于病因分层的特异靶点。

方法

实验对6周龄雄性C57小鼠给予高脂饮食连续喂养至18周龄，建立饮食诱导性肥胖（HFD）模型；同时选用18周龄雄性瘦素缺陷（ob/ob）和瘦素受体缺陷（db/db）遗传性肥胖小鼠，并以同龄雄性C57小鼠作为对照。每组取3只（

=3），共12只。处死后在体视显微镜下精细切取下丘脑组织。采用10x Genomics v3.1平台进行单细胞核转录组测序（snRNA-seq），数据经Cell Ranger比对后导入Seurat完成质控、降维与聚类分析；随后基于NeuronChat及其神经系统特异性配体-受体数据库推断神经元-神经元通讯网络，系统比较各肥胖模型与C57对照组在差异通讯网络、信号输出/输入强度、信息流及作用数量等方面的变化。

结果

单细胞转录组测序对C57、HFD、ob/ob及db/db小鼠下丘脑细胞进行降维聚类后，识别出包括兴奋性神经元、抑制性神经元及多类非神经元细胞在内的主要细胞群，不同模型同类细胞在UMAP空间存在一定偏移。对神经元二次聚类得到7个GABA能与10个谷氨酸能亚群，提示下丘脑神经元具有显著异质性。3种模型均呈现出GABA能抑制性输入和神经黏附分子-神经连接蛋白（Nrxn- Nlgn）通路信号普遍下调的特征，且不同模型间存在差异性神经通讯网络：HFD模型以缝隙连接蛋白α-1（Gja1-Gja1）电突触增强、Nrxn-Nlgn输出减少为特征；db/db模型以一氧化氮-鸟苷酸环化酶 1 可溶性亚基 alpha 2（NO-Gucy1a2）及强啡肽原（Pdyn）通路占优；ob/ob模型以谷氨酸受体互作增强为主。模型间两两比较进一步验证：HFD相对富集Gja1及部分代谢型谷氨酸受体互作，db/db相对富集NO-Gucy1a2及突触黏附相关通路，ob/ob相对富集离子型/代谢型谷氨酸受体互作。

结论

不同病因的肥胖模型中，下丘脑存在不同的神经通讯网络模式：饮食性肥胖更突出电突触相关通路增强，瘦素受体缺失模型以NO相关信号增强为主要特征，瘦素缺失模型则以谷氨酸能兴奋性互作增强为核心改变。但抑制性调控减弱是共同环节，为理解肥胖发生的病理机制提供了新的启示。

Abstract

Objective

To compare the hypothalamic cellular composition and neuron-neuron communication networks across 3 etiologically distinct mouse models of obesity

high-fat diet (HFD)

leptin-deficient (ob/ob)

and leptin receptor-deficient (db/db) so as to identify universal mechanistic clues that can be validated across etiologies and searching for etiology-specific targets applicable to stratified treatment.

Methods

Six-week-old male C57 mice were fed with HFD continuously until 18 weeks of age to establish a HFD model. In parallel

18-week-old male ob/ob mice and db/db mice were subjected as genetic obesity models

and age-matched male C57 mice served as controls. Three mice were included per group (

=3)

totaling 12 mice

were euthanized

and the hypothalamic tissues were meticulously dissected under a stereomicroscope. Single-nucleus transcriptome sequencing (snRNA-seq) was performed using the 10x Genomics v3.1 platform. The obtained sequencing data were aligned with Cell Ranger and processed in Seurat for quality control

dimensionality reduction

and clustering. Subsequently

neuron-neuron communication networks were inferred based on NeuronChat and its nervous system-specific ligand-receptor database

systematically comparing changes in differential communication networks

signal output/input strength

information flow

and interaction quantity between each obesity model and C57 controls.

Results

Dimensionality reduction and clustering of hypothalamic snRNA-seq data from C57

HFD

ob/ob

and db/db mice identified several major cell populations

including excitatory neurons

inhibitory neurons

and multiple non-neuronal cell types

with cells of the same type showing model-dependent shifts in uniform manifold approximation and projection (UMAP) space. Neuronal subclustering resolved 7 GABAergic and 10 glutamatergic subtypes

indicating pronounced neuronal heterogeneity in the hypothalamus. Across the 3 obese models

GABAergic inhibitory incoming signaling and the neurexin-neuroligin (Nrxn-Nlgn) pathway were broadly downregulated. Meanwhile

each model exhibited a distinct communication-network signature: the HFD model was characterized by enhanced electrical synapses mediated by gap junction protein alpha-1 (Gja1-Gja1) and reduced Nrxn-Nlgn output; the db/db model showed dominant nitric oxide-soluble guanylate cyclase signaling (NO-Gucy1a2) together with enrichment of prodynorphin (Pdyn)-related pathways; the ob/ob model was primarily marked by enhanced glutamate receptor interactions. Pairwise comparisons among models further validated these patterns: HFD relatively enriched Gja1-related signaling and certain metabotropic glutamate receptor interactions; db/db relatively enriched NO-Gucy1a2 and synaptic adhesion-related pathways; ob/ob relatively enriched in ionotropic/metabotropic glutamate receptor interactions.

Conclusion

Distinct neural communication network patterns exist in the hypothalamus across obesity models of different etiologies: diet-induced obesity preferentially exhibits enhanced electrical synapse-related pathways

leptin receptor-deficient model is primarily characterized by increased NO-related signaling

and the leptin-deficient model is centrally distinguished by enhanced glutamatergic excitatory interactions. Notably

attenuated inhibitory regulation emerges as a common feature across models

providing novel insights into understanding the pathological mechanisms underlying obesity development.

关键词

Keywords

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Xiaotang FAN

Yun CAI

Xin LI

Tianyao LIU

Ruotong RUAN

Rui XIAO

Lian WANG

Jinghui ZHAO

Related Institution

Department of Military Cognitive Psychology, Cdlege of Medical Psychology, Army Medical University (Third Military Medical University)

Department of Anesthesiology, Xinqiao Hospital, Third Military Medical University

Department of Developmental Neuropsychology, College of Psychology, Third Military Medical University

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