Department of Biochemistry, Institute of Health Sciences, Marmara University, Istanbul, Turkey.
Department of Biochemistry, Faculty of Medicine, Istanbul Medipol University, Istanbul, Turkey.
Methods Mol Biol. 2024;2753:447-457. doi: 10.1007/978-1-0716-3625-1_27.
Craniofacial abnormalities are one of the most frequent birth malformations in humans, affecting around one in every thousand live births. The zebrafish (Danio rerio), a model organism that has seen increased usage in toxicological research in recent years, is ideal for assessing the effects of various chemicals on bone and cartilage structures. Chondrogenesis developed in zebrafish embryos by embryonic day 2, and supporting cartilage components are apparent at hatching (72 h post-fertilization). Individual cartilage may be observed using Alcian Blue staining as early as 2 days post-fertilization (dpf). The preferential binding of Alcian Blue causes the staining of zebrafish cartilage to acidic glycoproteins in an acidic solution (pH 2.2). In 72-120 hpf embryos, the cranial skeleton is easily visible after cartilage staining using Alcian Blue. Various cranial lengths and structures can be determined by measuring specific distances and angles to optimize the quantitative analysis of cranial malformations in zebrafish after exposure to various toxic agents. This chapter explains the Alcian Blue staining procedure to identify craniofacial cartilaginous structures in zebrafish embryos.
颅面畸形是人类最常见的出生缺陷之一，影响大约每千例活产中的一例。斑马鱼（Danio rerio）是一种近年来在毒理学研究中越来越多地被使用的模式生物，非常适合评估各种化学物质对骨骼和软骨结构的影响。斑马鱼胚胎在胚胎发育第 2 天开始软骨生成，孵化时（受精后 72 小时）可见支持软骨的成分。早在受精后 2 天（dpf），就可以使用 Alcian Blue 染色观察单个软骨。在酸性溶液（pH 2.2）中，Alcian Blue 优先与酸性糖蛋白结合，从而使斑马鱼软骨染色。在 72-120 hpf 胚胎中，软骨染色后很容易看到颅骨骼。通过测量特定的距离和角度，可以确定各种颅长度和结构，以优化暴露于各种有毒剂后的斑马鱼颅面畸形的定量分析。本章介绍了 Alcian Blue 染色程序，用于鉴定斑马鱼胚胎中的颅面软骨结构。
