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来自寨卡病毒感染的斑马鱼胚胎的神经祖细胞的定量分析。

Quantification of Neural Progenitor Cells From Zika Virus-Infected Zebrafish Embryos.

作者信息

Gomes Yago C P, Sow Aïssatou Aïcha, Patten Shunmoogum A, Chatel-Chaix Laurent

机构信息

Centre Armand-Frappier Santé Biotechnologie, Institut National de la Recherche Scientifique, Laval, QC, Canada.

出版信息

Bio Protoc. 2025 Jul 5;15(13):e5377. doi: 10.21769/BioProtoc.5377.

Abstract

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.

摘要

寨卡病毒(ZIKV)是一种节肢动物传播的正黄病毒属病毒,因其能够导致严重的胎儿神经紊乱,进而在新生儿中引发先天性寨卡综合征(CZS),已成为全球关注的健康问题。孕期的垂直传播会改变神经祖细胞(NPC)的增殖和分化,并诱导细胞凋亡,导致小头畸形和其他神经发育异常。虽然已使用哺乳动物模型来研究ZIKV对NPC行为的影响,但诸如高成本、特定时间和伦理限制等局限性促使人们探索替代系统。斑马鱼胚胎构成了研究ZIKV神经发病机制的一种有利的体内模型。实际上,ZIKV感染模拟了CZS的几个特征,同时具有保守的神经解剖布局,并且与哺乳动物相比,为受感染的大脑提供了遗传可塑性和独特的可及性。在此,我们描述了一种利用全动物流式细胞术在这种斑马鱼模型中表征ZIKV诱导的NPC缺陷的方案。关键特征 • 本方案引入了一种新颖且简单的基于斑马鱼的系统,用于研究ZIKV诱导的神经发病机制。 • 该方案采用一种无偏倚的方法,通过流式细胞术对整个胚胎中的神经祖细胞进行定量。 • 本方案可应用于任何产生特定荧光细胞群体的转基因报告斑马鱼品系。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f63/12245632/2fa69bc9c809/BioProtoc-15-13-5377-g001.jpg

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