Proteome profile differences among human, monkey, and mouse brain microvessels and cultured brain microvascular endothelial cells.

IF 5.9 1区医学 Q1 NEUROSCIENCES

Fluids and Barriers of the CNS Pub Date : 2025-05-30 DOI:10.1186/s12987-025-00650-z

Haruka Kumabe, Takeshi Masuda, Shingo Ito, Tomomi Furihata, Akiko Toda, Masayuki Mogi, Norie Araki, Sumio Ohtsuki

{"title":"Proteome profile differences among human, monkey, and mouse brain microvessels and cultured brain microvascular endothelial cells.","authors":"Haruka Kumabe, Takeshi Masuda, Shingo Ito, Tomomi Furihata, Akiko Toda, Masayuki Mogi, Norie Araki, Sumio Ohtsuki","doi":"10.1186/s12987-025-00650-z","DOIUrl":null,"url":null,"abstract":"Background: The blood-brain barrier (BBB) expresses transporters, receptors, and tight junction proteins that regulate the exchange of substances between the blood and brain. The differences in the expression of these proteins in the BBB among different species and cultured BBB model cells should be clarified to interpret the BBB function in model animals and cells. This study aimed to elucidate species differences among humans, monkeys, and mice and in vitro-in vivo differences in the BBB proteome using deep proteomic analysis.Methods: Brain microvessels (BMVs) were isolated from frozen cerebral cortices of human and monkey and frozen mouse cerebrums. BMVs and cultured brain microvascular endothelial cells (BMECs), such as hCMEC/D3, HBMEC/ciβ, and primary BMECs, were analyzed via data-independent acquisition using liquid chromatography-mass spectrometry.Results: Proteomics identified 7,149-8,274 proteins in the BMV fractions and 6,657-7,534 proteins in the brain lysates. Comparative analysis revealed distinct proteomic profiles among the three species, with the human profile being more similar to that of monkeys than that of mice. The expression profile of the solute carrier organic anion transporter family was found to vary among mouse, monkey, and human BMVs. The expression levels of SLC22A6/Slc22a6 and SLC22A8/Slc22a8 were higher in mice than in monkeys and humans, whereas SLC43A3/Slc43a3 expression levels were lower in mice. The expression of amino acid transporters, such as SLC7A5 and SLC3A2/Slc3a2, was higher in BMVs, whereas that of SLC1A5/Slc1a5 and SLC38A9/Slc38a9 was higher in cultured BMECs. MFSD2A/Mfsd2a and SLC27A1/Slc27a1 were highly expressed in BMVs. The expression of tight junction proteins, particularly the claudin family, varied between BMVs and cultured BMECs and among cell lines. Specifically, the expression of claudin-5 was higher in BMVs, and claudin-11 expression was higher in cultured BMECs.Conclusions: Deep proteomic analysis revealed species-specific differences in transport-related proteins in the BBB. Furthermore, in vitro and in vivo differences were observed in the transporter and claudin protein expression. This study provides a BBB proteome profile dataset and offers insights for a comprehensive understanding of BBB protein expression across species and between in vivo and in vitro conditions.","PeriodicalId":12321,"journal":{"name":"Fluids and Barriers of the CNS","volume":"22 1","pages":"53"},"PeriodicalIF":5.9000,"publicationDate":"2025-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12124085/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Fluids and Barriers of the CNS","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1186/s12987-025-00650-z","RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"NEUROSCIENCES","Score":null,"Total":0}

引用次数: 0

Abstract

Background: The blood-brain barrier (BBB) expresses transporters, receptors, and tight junction proteins that regulate the exchange of substances between the blood and brain. The differences in the expression of these proteins in the BBB among different species and cultured BBB model cells should be clarified to interpret the BBB function in model animals and cells. This study aimed to elucidate species differences among humans, monkeys, and mice and in vitro-in vivo differences in the BBB proteome using deep proteomic analysis.

Methods: Brain microvessels (BMVs) were isolated from frozen cerebral cortices of human and monkey and frozen mouse cerebrums. BMVs and cultured brain microvascular endothelial cells (BMECs), such as hCMEC/D3, HBMEC/ciβ, and primary BMECs, were analyzed via data-independent acquisition using liquid chromatography-mass spectrometry.

Results: Proteomics identified 7,149-8,274 proteins in the BMV fractions and 6,657-7,534 proteins in the brain lysates. Comparative analysis revealed distinct proteomic profiles among the three species, with the human profile being more similar to that of monkeys than that of mice. The expression profile of the solute carrier organic anion transporter family was found to vary among mouse, monkey, and human BMVs. The expression levels of SLC22A6/Slc22a6 and SLC22A8/Slc22a8 were higher in mice than in monkeys and humans, whereas SLC43A3/Slc43a3 expression levels were lower in mice. The expression of amino acid transporters, such as SLC7A5 and SLC3A2/Slc3a2, was higher in BMVs, whereas that of SLC1A5/Slc1a5 and SLC38A9/Slc38a9 was higher in cultured BMECs. MFSD2A/Mfsd2a and SLC27A1/Slc27a1 were highly expressed in BMVs. The expression of tight junction proteins, particularly the claudin family, varied between BMVs and cultured BMECs and among cell lines. Specifically, the expression of claudin-5 was higher in BMVs, and claudin-11 expression was higher in cultured BMECs.

Conclusions: Deep proteomic analysis revealed species-specific differences in transport-related proteins in the BBB. Furthermore, in vitro and in vivo differences were observed in the transporter and claudin protein expression. This study provides a BBB proteome profile dataset and offers insights for a comprehensive understanding of BBB protein expression across species and between in vivo and in vitro conditions.

查看原文本刊更多论文

人、猴和小鼠脑微血管和培养的脑微血管内皮细胞的蛋白质组谱差异。

背景：血脑屏障（BBB）表达转运蛋白、受体和紧密连接蛋白，这些蛋白调节血液和大脑之间的物质交换。研究这些蛋白在不同物种和培养的血脑屏障模型细胞中在血脑屏障中的表达差异，有助于解释血脑屏障在模型动物和细胞中的功能。本研究旨在通过深度蛋白质组学分析阐明人类、猴子和小鼠之间的物种差异以及血脑屏障蛋白质组的体内外差异。方法：从冷冻的人、猴和小鼠大脑皮层中分离脑微血管。采用液相色谱-质谱法对bmv和培养的脑微血管内皮细胞（BMECs），如hCMEC/D3、HBMEC/ciβ和原代BMECs进行数据独立采集分析。结果：蛋白质组学在BMV中鉴定出7,149-8,274个蛋白质，在脑裂解物中鉴定出6,657-7,534个蛋白质。对比分析显示，这三个物种之间的蛋白质组学特征不同，人类的蛋白质组学特征与猴子的更相似，而与老鼠的更相似。发现溶质载体有机阴离子转运蛋白家族在小鼠、猴和人bmv中的表达谱存在差异。SLC22A6/ SLC22A6和SLC22A8/ SLC22A8在小鼠中的表达水平高于猴子和人，而SLC43A3/ SLC43A3在小鼠中的表达水平较低。氨基酸转运体SLC7A5和SLC3A2/ SLC3A2在bmmes中表达量较高，而SLC1A5/ SLC1A5和SLC38A9/ SLC38A9在bmmes中表达量较高。MFSD2A/ MFSD2A和SLC27A1/ SLC27A1在bmv中高表达。紧密连接蛋白的表达，特别是claudin家族，在bmv和培养的bmec之间以及细胞系之间存在差异。其中，claudin-5在bmv中表达较高，claudin-11在bmc中表达较高。结论：深层蛋白质组学分析揭示了血脑屏障中运输相关蛋白的物种特异性差异。此外，在体外和体内观察到转运蛋白和claudin蛋白表达的差异。该研究提供了一个血脑屏障蛋白质组数据集，并为全面了解血脑屏障蛋白在物种之间以及体内和体外条件下的表达提供了见解。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Fluids and Barriers of the CNS Neuroscience-Developmental Neuroscience

CiteScore

10.70

自引率

8.20%

发文量

审稿时长

14 weeks

期刊介绍： "Fluids and Barriers of the CNS" is a scholarly open access journal that specializes in the intricate world of the central nervous system's fluids and barriers, which are pivotal for the health and well-being of the human body. This journal is a peer-reviewed platform that welcomes research manuscripts exploring the full spectrum of CNS fluids and barriers, with a particular focus on their roles in both health and disease. At the heart of this journal's interest is the cerebrospinal fluid (CSF), a vital fluid that circulates within the brain and spinal cord, playing a multifaceted role in the normal functioning of the brain and in various neurological conditions. The journal delves into the composition, circulation, and absorption of CSF, as well as its relationship with the parenchymal interstitial fluid and the neurovascular unit at the blood-brain barrier (BBB).