Characterizing newly repopulated microglia in the adult mouse: impacts on animal behavior, cell morphology, and neuroinflammation.
Characterizing newly repopulated microglia in the adult mouse: impacts on animal behavior, cell morphology, and neuroinflammation.
复制标题
DOI：
10.1371/journal.pone.0122912
复制
发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Green KN
中科院分区：
文献类型：
--
作者：
Elmore MR;Lee RJ;West BL;Green KN
小胶质细胞是大脑中的主要免疫细胞，被认为在免疫之外发挥重要作用。对成年小鼠给予双重集落刺激因子1受体（CSF 1 R）/c-Kit激酶抑制剂PLX 3397可消除约99%的小胶质细胞，只要治疗继续，小胶质细胞就保持消除状态。在去除抑制剂后，小胶质细胞迅速重新填充整个成人大脑，源于中枢神经系统（CNS）驻留祖细胞。使用这种小胶质细胞消除和再增殖的方法，可以探索小胶质细胞在健康和疾病状态中的作用。在这里，我们研究了新重新填充的小胶质细胞对炎症刺激的反应，以及确定这些细胞对行为，认知和神经炎症的影响。将两个月大的野生型小鼠置于对照或PLX 3397饮食21天以消除小胶质细胞。然后在一个动物亚组中去除PLX 3397饮食，以允许小胶质细胞重新增殖，并在14天重新增殖时开始进行行为测试。最后，使用定量真实的时间聚合酶链反应（RT-PCR）以及小胶质细胞形态学的详细分析，在去除抑制剂后21天确定小胶质细胞再填充脑响应于脂多糖（LPS; 0.25 mg/kg）或磷酸盐缓冲盐水（PBS）的炎症谱。我们发现，通过行为、认知和运动功能的测量，重新填充小胶质细胞的小鼠表现与对照组相似。与对照/常驻小胶质细胞相比，重新填充的小胶质细胞具有更大的细胞体和更少的复杂分支，其在抑制剂去除后随时间推移而消退。炎症分析显示，重新填充的小胶质细胞的mRNA基因表达与正常的常驻小胶质细胞相似，并且这些新细胞表现出功能性并对LPS有反应。总体而言，这些数据表明，新重新填充的小胶质细胞的功能与原来的居民小胶质细胞相似，在健康成年小鼠中没有任何明显的不良影响。
Microglia are the primary immune cell in the brain and are postulated to play important roles outside of immunity. Administration of the dual colony-stimulating factor 1 receptor (CSF1R)/c-Kit kinase inhibitor, PLX3397, to adult mice results in the elimination of ~99% of microglia, which remain eliminated for as long as treatment continues. Upon removal of the inhibitor, microglia rapidly repopulate the entire adult brain, stemming from a central nervous system (CNS) resident progenitor cell. Using this method of microglial elimination and repopulation, the role of microglia in both healthy and diseased states can be explored. Here, we examine the responsiveness of newly repopulated microglia to an inflammatory stimulus, as well as determine the impact of these cells on behavior, cognition, and neuroinflammation. Two month-old wild-type mice were placed on either control or PLX3397 diet for 21 d to eliminate microglia. PLX3397 diet was then removed in a subset of animals to allow microglia to repopulate and behavioral testing conducted beginning at 14 d repopulation. Finally, inflammatory profiling of the microglia-repopulated brain in response to lipopolysaccharide (LPS; 0.25 mg/kg) or phosphate buffered saline (PBS) was determined 21 d after inhibitor removal using quantitative real time polymerase chain reaction (RT-PCR), as well as detailed analyses of microglial morphologies. We find mice with repopulated microglia to perform similarly to controls by measures of behavior, cognition, and motor function. Compared to control/resident microglia, repopulated microglia had larger cell bodies and less complex branching in their processes, which resolved over time after inhibitor removal. Inflammatory profiling revealed that the mRNA gene expression of repopulated microglia was similar to normal resident microglia and that these new cells appear functional and responsive to LPS. Overall, these data demonstrate that newly repopulated microglia function similarly to the original resident microglia without any apparent adverse effects in healthy adult mice.
登录
查看更多内容
影响因子：
13.5
作者：
Harry, G. Jean
通讯作者：
Harry, G. Jean
影响因子：
15.1
作者：
Chen, Jing;Buchanan, Jessica B.;Johnson, Rodney W.
通讯作者：
Johnson, Rodney W.
影响因子：
5.7
作者：
Coniglio, Salvatore J.;Eugenin, Eliseo;Segall, Jeffrey E.
通讯作者：
Segall, Jeffrey E.
影响因子：
6.2
作者：
Blank, Thomas;Prinz, Marco
通讯作者：
Prinz, Marco
影响因子：
3
作者：
Bilbo SD;Schwarz JM
通讯作者：
Schwarz JM