New tools for studying microglia in the mouse and human CNS
New tools for studying microglia in the mouse and human CNS
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DOI：
10.1073/pnas.1525528113
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发表时间：
2016-03-22
影响因子：
11.1
通讯作者：
Barres, Ben A.
中科院分区：
文献类型：
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作者：
Bennett, Mariko L.;Bennett, F. Chris;Barres, Ben A.
由于缺乏区分小胶质细胞和其他免疫细胞的工具，因此难以确定小胶质细胞（脑和脊髓的组织驻留巨噬细胞）的特定功能，从而限制了特异性免疫染色、纯化和操作。由于它们独特的发育起源和预测功能，小胶质细胞与其他髓样细胞的区别对于理解脑发育和疾病至关重要;更好的工具将大大促进小胶质细胞在发育中，成年和受损CNS中功能的研究。在这里，我们确定跨膜蛋白119（Tmem 119），一个未知功能的细胞表面蛋白，作为一个高度表达的小胶质细胞特异性标记在小鼠和人类。我们开发了其细胞内和细胞外结构域的单克隆抗体，使健康和患病的大脑组织切片中的小胶质细胞的免疫染色，以及通过流式细胞仪分离纯非活化的小胶质细胞。据我们所知，使用我们的抗体，我们提供了在发育过程中和免疫挑战后高纯度小鼠小胶质细胞的第一个RNAseq图谱。我们用这些来证明小鼠小胶质细胞在出生后第二周成熟，并预测新的小胶质细胞功能。总之，我们预计这些资源将是宝贵的未来研究和了解小胶质细胞在健康和疾病。
The specific function of microglia, the tissue resident macrophages of the brain and spinal cord, has been difficult to ascertain because of a lack of tools to distinguish microglia from other immune cells, thereby limiting specific immunostaining, purification, and manipulation. Because of their unique developmental origins and predicted functions, the distinction of microglia from other myeloid cells is critically important for understanding brain development and disease; better tools would greatly facilitate studies of microglia function in the developing, adult, and injured CNS. Here, we identify transmembrane protein 119 (Tmem119), a cell-surface protein of unknown function, as a highly expressed microglia-specific marker in both mouse and human. We developed monoclonal antibodies to its intracellular and extracellular domains that enable the immunostaining of microglia in histological sections in healthy and diseased brains, as well as isolation of pure nonactivated microglia by FACS. Using our antibodies, we provide, to our knowledge, the first RNAseq profiles of highly pure mouse microglia during development and after an immune challenge. We used these to demonstrate that mouse microglia mature by the second postnatal week and to predict novel microglial functions. Together, we anticipate these resources will be valuable for the future study and understanding of microglia in health and disease.