A genetic tool for the longitudinal study of a subset of post-inflammatory reactive astrocytes.
A genetic tool for the longitudinal study of a subset of post-inflammatory reactive astrocytes.
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DOI：
10.1016/j.crmeth.2022.100276
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发表时间：
2022-08-22
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星形胶质细胞是保证大脑正常功能的重要支持细胞。在脑部疾病中，星形胶质细胞重编程进入反应状态，改变了它们的许多细胞功能。该领域一个长期存在的问题是，炎症消退过程中反应性星形胶质细胞（RA）标记物的下调是否因为这些星形胶质细胞恢复到非反应状态或死亡并被替换。事实证明，这很难回答，主要是因为现有的遗传工具无法区分健康和RAs。在这里，我们描述了可用于特异性靶向和标记RAs子集的诱导遗传工具的生成。使用该工具对急性炎症模型进行的纵向分析显示，先前观察到的炎症后RA标记物的下调可能是由于基因表达的改变，而不是由于细胞死亡。我们的研究结果表明，急性炎症后与星形胶质细胞增生相关的细胞变化在很大程度上是可逆的。Cre在多种脑炎症模型的RAs中表达，可以对RAs进行永久性标记以进行纵向分析。在急性炎症后1个月，RAs基本上会恢复到非反应状态。对反应性星形胶质细胞（RAs）的体内研究已被证明是困难的，主要是因为现有的使用传统星形胶质细胞启动子的遗传工具无法区分健康和反应性星形胶质细胞。为了填补这一空白，我们描述了一种可诱导遗传工具的产生和表征，该工具可用于特异性靶向、标记和操纵病变或损伤大脑中的RAs子集。我们为这种方法在几种脑炎症模型中研究RAs的效用提供了证据。Agnew-Svoboda等人生成了一种可诱导的Cre系，能够选择性靶向和操纵脑疾病中的反应性星形胶质细胞（RAs）子集。对RAs的纵向分析显示，急性炎症后迅速反应的星形胶质细胞存活下来，但一旦炎症消退，它们大部分会恢复到非反应状态。
Astrocytes are vital support cells that ensure proper brain function. In brain disease, astrocytes reprogram into a reactive state that alters many of their cellular roles. A long-standing question in the field is whether downregulation of reactive astrocyte (RA) markers during resolution of inflammation is because these astrocytes revert back to a non-reactive state or die and are replaced. This has proven difficult to answer mainly because existing genetic tools cannot distinguish between healthy versus RAs. Here we describe the generation of an inducible genetic tool that can be used to specifically target and label a subset of RAs. Longitudinal analysis of an acute inflammation model using this tool revealed that the previously observed downregulation of RA markers after inflammation is likely due to changes in gene expression and not because of cell death. Our findings suggest that cellular changes associated with astrogliosis after acute inflammation are largely reversible. An inducible Cre line to selectively target a subset of reactive astrocytes (RAs) Cre is expressed in RAs across a wide array of brain inflammation models Enables the permanent tagging of RAs for longitudinal analysis RAs largely revert to a non-reactive state 1 month after acute inflammation In vivo study of reactive astrocytes (RAs) has proven difficult mainly because existing genetic tools using traditional astrocyte promoters cannot distinguish between healthy versus reactive astrocytes. To fill this gap, we describe the generation and characterization of an inducible genetic tool that can be used to specifically target, label, and manipulate a subset of RAs in the diseased or injured brain. We provide evidence for the utility of this approach to study RAs in several models of brain inflammation. Agnew-Svoboda et al. generate an inducible Cre line to enable selective targeting and manipulation of a subset of reactive astrocytes (RAs) in brain disease. Longitudinal analysis of RAs reveal that astrocytes that become reactive soon after acute inflammation survive but largely revert to a non-reactive state once inflammation has resolved.