小鼠视网膜和大脑边界组织中的组织特异性免疫转录信号

IF 5 2区 医学 Q1 OPHTHALMOLOGY Investigative ophthalmology & visual science Pub Date : 2024-10-01 DOI:10.1167/iovs.65.12.42
Fazeleh Etebar, Paul Whatmore, Damien G Harkin, Samantha J Dando
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引用次数: 0

摘要

目的:与中枢神经系统(CNS)实质相邻的是脉络膜(视网膜下层)和脑膜(包裹大脑的脑膜内层)。虽然脉络膜和脑膜靠近神经实质,但它们处于视网膜和大脑的免疫环境之外,因此是不同的免疫区。本研究旨在描述健康成年小鼠中枢神经系统边缘脉络膜和脑膜内免疫细胞的转录组特征:方法:从7周大的C57Bl/6J小鼠身上获取眼球和大脑。对脉络膜和脑膜进行处理,以分离 CD45+ 免疫细胞和进行单细胞 RNA 测序。此外,还对从供体眼组织脉络膜中分离出的免疫细胞进行了单细胞 RNA 序列分析。使用免疫染色和共聚焦显微镜对整块组织进行原位验证所选的免疫细胞群:结果:共对小鼠组织中的 3606 个细胞进行了测序,包括来自脉络膜的 2125 个 CD45+ 细胞和来自脑膜的 1481 个 CD45+ 细胞。聚类和差异基因表达分析显示了单核细胞/巨噬细胞、树突状细胞、T细胞和B细胞的不同亚型。有些集群在脉络膜和脑膜中常见,而其他集群则表现出组织特异性基因表达谱和潜在的功能特化。对来自人类脉络膜的 6501 个 CD45+ 细胞进行测序分析,发现了与小鼠脉络膜相似的免疫细胞群:这项研究详细描述了健康视网膜和大脑周围血管结缔组织中免疫细胞的分子特征及其在免疫保护中的潜在作用。
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Tissue-Specific Immune Transcriptional Signatures in the Bordering Tissues of the Mouse Retina and Brain.

Purpose: Bordering the central nervous system (CNS) parenchyma are the choroid (underlying the retina) and the leptomeninges (the inner layers of the meninges enveloping the brain). Although near the neural parenchyma, the choroid and leptomeninges are external to the immune privileged environment of the retina and brain and thus are distinct immune compartments. This study aimed to characterize the transcriptomic signatures of immune cells within the choroid and leptomeninges bordering the healthy adult mouse CNS.

Methods: Eyes and brains were obtained from 7-week-old C57Bl/6J mice. Choroid and leptomeninges were processed for isolation of CD45+ immune cells and single cell RNA-sequencing. Additionally, single cell RNA-sequencing was performed on immune cells isolated from choroid obtained from human donor eye tissue. Immunostaining and confocal microscopy of wholemount tissue were used to validate selected immune cell populations in situ.

Results: A total of 3606 cells were sequenced from mouse tissues, including 2125 CD45+ cells from choroid and 1481 CD45+ cells from leptomeninges. Clustering and differential gene expression analysis revealed heterogeneous subtypes of monocytes/macrophages, dendritic cells, T cells, and B cells. Whereas some clusters were common to both choroid and leptomeninges, others exhibited tissue-specific gene expression profiles and potential functional specializations. Analysis of 6501 CD45+ cells sequenced from human choroid identified similar immune cell populations to mouse choroid.

Conclusions: This study provides a detailed characterization of the molecular signatures of immune cells within the vascular connective tissues bordering the healthy retina and brain, and their potential roles in immune protection.

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来源期刊
CiteScore
6.90
自引率
4.50%
发文量
339
审稿时长
1 months
期刊介绍: Investigative Ophthalmology & Visual Science (IOVS), published as ready online, is a peer-reviewed academic journal of the Association for Research in Vision and Ophthalmology (ARVO). IOVS features original research, mostly pertaining to clinical and laboratory ophthalmology and vision research in general.
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