Single-cell analysis of murine long-term hematopoietic stem cells reveals distinct patterns of gene expression during fetal migration.
Single-cell analysis of murine long-term hematopoietic stem cells reveals distinct patterns of gene expression during fetal migration.
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DOI：
10.1371/journal.pone.0030542
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
García-Ojeda ME
中科院分区：
文献类型：
--
作者：
Ciriza J;Hall D;Lu A;De Sena JR;Al-Kuhlani M;García-Ojeda ME
长期造血干细胞(LT-HSCs)在发育过程中从胎肝(FL)迁移到胎儿骨髓(FBM)。多种黏附和趋化受体基因参与了成体LT-HSCs的迁移。然而，它们在胎儿LT-HSCs迁移中的作用尚不清楚，部分原因是这些细胞在胎儿组织中的数量很少，这排除了经典的基因表达分析。本研究的目的是研究迁移相关基因在发育过程中不同解剖位置的胎儿LT-HSC中的表达。我们从不同发育阶段和不同解剖位置分离了胎儿LT-HSC，并对参与成人LT-HSC迁移的8个分子进行了单细胞多重RT-qPCR和流式细胞术分析。我们的结果表明，趋化因子受体CXCR4在LT-HSC中的基因表达随发育微环境和发育时间的不同而不同，而钙粘附素CDH2(Ncad)和钙受体CaSR仅在胚胎发育后17.5天(DPC)表现出较高的基因表达和变异性。钙粘蛋白CDH5(Vecad)仅在胚胎发育过程中保持较高的表达变异性，而整合素亚单位ITGA5(α5)在14.5dpc后表达的变异性增加。整合素亚单位ItGa4(α4)和ItGal(Lfa1)以及选择素配体Selplg(Psgl1)在单个LT-HSC中的表达没有差异，与研究的发育时间和解剖微环境无关。我们的数据表明，表型相同的单个LT-HSCs的表达模式随着发育阶段和解剖微环境的变化而波动。这是第一次对不同发育时期胎儿组织中迁移相关分子的基因表达进行详尽的比较，增强了对LT-HSC在发育过程中迁移命运决定的理解。
Long-term hematopoietic stem cells (LT-HSCs) migrate from the fetal liver (FL) to the fetal bone marrow (FBM) during development. Various adhesion and chemotactic receptor genes have been implicated in the migration of adult LT-HSCs. However, their role in the migration of fetal LT-HSCs is not clearly understood due, in part, to the rare number of these cells in fetal tissues, which preclude classical gene expression analysis. The aim of this study is to characterize the expression of migration related genes in fetal LT-HSC across different anatomical locations during development. We isolated fetal LT-HSC from different developmental stages, as well as different anatomical locations, and performed single-cell multiplex RT-qPCR and flow cytometry analysis of eight molecules involved in adult LT-HSC migration. Our results show that the gene expression of the chemokine receptor Cxcr4 in LT-HSC varies across developmental microenvironments and times, while the cadherin Cdh2 (Ncad) and the calcium receptor Casr show higher gene expression and variability only in FBM at 17.5 days post coitum (dpc). The cadherin Cdh5 (Vecad) maintains high expression variability only during fetal development, while the integrin subunit Itga5 (α5) increases its variability after 14.5 dpc. The integrin subunits Itga4 (α4) and Itgal (Lfa1), as well as the selectin ligand Selplg (Psgl1), did not show differences in their expression in single LT-HSCs irrespective of the developmental times or anatomical microenvironments studied. Our data demonstrate that the expression pattern of phenotypically identical, single LT-HSCs fluctuates as a function of developmental stage and anatomical microenvironment. This is the first exhaustive gene expression comparison of migration-related molecules in fetal tissues across developmental times, enhancing the understanding of LT-HSC migration fate decisions during development.
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影响因子：
64.5
作者：
Kiel, MJ;Yilmaz, ÖH;Morrison, SJ
通讯作者：
Morrison, SJ
影响因子：
20.3
作者：
Hosokawa, Kentaro;Arai, Fumio;Suda, Toshio
通讯作者：
Suda, Toshio
影响因子：
64.8
作者：
Chan, Charles K. F.;Chen, Ching-Cheng;Luppen, Cynthia A.;Kim, Jae-Beom;DeBoer, Anthony T.;Wei, Kevin;Helms, Jill A.;Kuo, Calvin J.;Kraft, Daniel L.;Weissman, Irving L.
通讯作者：
Weissman, Irving L.
影响因子：
32.4
作者：
Ara, T;Tokoyoda, K;Nagasawa, T
通讯作者：
Nagasawa, T
DOI：
10.1073/pnas.95.24.14423
发表时间：
1998-11-24
影响因子：
11.1
作者：
Frenette, PS;Subbarao, S;Wagner, DD
通讯作者：
Wagner, DD