寨卡病毒感染斑马鱼胚胎神经祖细胞的定量分析。

IF 1 Q3 BIOLOGY
Yago C P Gomes, Aïssatou Aïcha Sow, Shunmoogum A Patten, Laurent Chatel-Chaix
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引用次数: 0

摘要

寨卡病毒(ZIKV)是一种节肢动物传播的正黄病毒,由于其能够引起严重的胎儿神经系统疾病,导致新生儿出现先天性寨卡综合征(CZS),已成为全球卫生问题。妊娠期垂直传播可改变神经祖细胞(NPC)的增殖和分化,诱导细胞凋亡,导致小头畸形等神经发育异常。虽然哺乳动物模型已被用于研究寨卡病毒对NPC行为的影响,但诸如高成本、专用时间和道德约束等限制促使人们探索替代系统。斑马鱼胚胎是研究寨卡病毒神经发病机制的有利体内模型。事实上,与哺乳动物相比,寨卡病毒感染表现了CZS的几个特征,同时共享保守的神经解剖布局,并提供遗传可塑性和受感染大脑的独特可及性。在这里,我们描述了一种基于全动物流式细胞术的方案,用于表征斑马鱼模型中寨卡病毒诱导的npc缺陷。•该协议引入了一种新颖而直接的基于斑马鱼的系统,用于研究zikv诱导的神经发病机制。•该方案采用无偏方法学流式细胞术为基础的神经祖细胞全胚胎定量。•该方案可以转位到任何转基因报告斑马鱼系产生特定的荧光细胞群。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Quantification of Neural Progenitor Cells From Zika Virus-Infected Zebrafish Embryos.

Zika virus (ZIKV), an arthropod-borne orthoflavivirus, has emerged as a global health concern due to its ability to cause severe fetal neurological disorders, leading to the congenital Zika syndrome (CZS) in neonates. Vertical transmission during pregnancy can alter neural progenitor cell (NPC) proliferation and differentiation and induce apoptosis, leading to microcephaly and other neurodevelopmental abnormalities. While mammalian models have been used to study the impact of ZIKV on NPC behavior, limitations such as high costs, dedicated time, and ethical constraints have fostered the exploration of alternative systems. The zebrafish embryo constitutes an advantageous in vivo model for studying ZIKV neuropathogenesis. Indeed, ZIKV infection phenocopies several features of the CZS while sharing a conserved neuroanatomical layout and offering genetic plasticity and unique accessibility to the infected brain compared to mammals. Here, we describe a protocol for characterizing ZIKV-induced defects of NPCs in this zebrafish model, relying on whole animal flow cytometry. Key features • This protocol introduces a novel and straightforward zebrafish-based system for studying ZIKV-induced neuropathogenesis. • The protocol employs an unbiased methodology for flow cytometry-based quantification of neural progenitor cells from whole embryos. • This protocol can be transposed to any transgenic reporter zebrafish lines producing specific fluorescent cell populations.

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