A Drug-Inducible Transgenic Zebrafish Model for Myelinating Glial Cell Ablation.
A Drug-Inducible Transgenic Zebrafish Model for Myelinating Glial Cell Ablation.
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DOI：
10.1007/978-1-4939-9072-6_13
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发表时间：
2019
影响因子：
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通讯作者：
Marja J Karttunen;D. Lyons
中科院分区：
文献类型：
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作者：
Marja J Karttunen;D. Lyons
为了研究体内脱髓鞘和髓鞘再生的细胞和分子机制，我们开发了一种转基因斑马鱼系Tg（mbp：mCherry-NTR），其中细菌酶硝基还原酶（NTR）的表达在髓鞘碱性蛋白启动子（mbp）下驱动，从而在髓鞘形成神经胶质中表达。当用前药甲硝唑处理表达NTR的幼虫时，NTR和Mtz之间的反应产生选择性杀死表达NTR的细胞的毒性代谢物。使用Tg（mbp：mCherry-NTR）系，我们可以在2天MTZ处理后消融三分之二的少突胶质细胞。脱髓鞘在7天后是明显的，并且在Mtz处理后16天观察到髓鞘再生。Tg（mbp：mCherry-NTR）模型可用于在脱髓鞘和髓鞘再生期间对细胞行为进行成像，并且用于测试遗传操作或化学化合物如何调节脱髓鞘和髓鞘再生。在这一章中，我们描述了我们用来描述少突胶质细胞损失，脱髓鞘和髓鞘再生的Tg（mbp：mCherry-NTR）模型的方法。
To study cellular and molecular mechanisms of demyelination and remyelination in vivo, we developed a transgenic zebrafish line, Tg(mbp:mCherry-NTR), in which expression of the bacterial enzyme nitroreductase (NTR) is driven under the myelin basic protein promoter (mbp) and thus is expressed in myelinating glia. When NTR-expressing larvae are treated with the prodrug metronidazole, the reaction between NTR and Mtz results in a toxic metabolite which selectively kills NTR-expressing cells. Using the Tg(mbp:mCherry-NTR) line, we can ablate two-thirds of oligodendrocytes following a 2-day MTZ treatment. Demyelination is evident seven days later, and remyelination is observed 16 days after Mtz treatment. The Tg(mbp:mCherry-NTR) model can be used to image cell behavior during, and to test how genetic manipulations or chemical compounds regulate, demyelination and remyelination. In this chapter, we describe the methods we used to characterize the oligodendrocyte loss, demyelination and remyelination in the Tg(mbp:mCherry-NTR) model.