M Florencia Sampedro, Marta E Sian, Paulo E Cabral Filho, Adriana Fontes, Valeria Sigot
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引用次数: 0
摘要
斑马鱼表皮是一个可接近的组织,可用于评估药物对上皮完整性的影响。虽然乙二醇-双(β-氨基乙醚)-N,N,N‘,N’-四乙酸(EGTA E3889, Sigma-Aldrich, CABA-Argentina)是一种钙螯合剂,在体外被广泛用于破坏上皮单层,但其在斑马鱼体内的应用仍然有限。我们发现,短期暴露于EGTA会导致尾巴损伤,从轻微和可逆到严重和致命;在组织水平上,它破坏了钙粘蛋白介导的细胞间接触和表皮的屏障功能。荧光量子点(QDs)标记能够快速表征损伤严重程度,证明包膜层中肌动蛋白微晶的消失。暴露于25 mM EGTA至少15分钟,可在咽期诱导可逆性上皮破坏,为研究斑马鱼胚胎的愈合和再生提供化学模型。
EGTA-induced disruption of adherens junctions in zebrafish embryonic epidermis.
Zebrafish epidermis is an accessible tissue to evaluate the impact of drugs on epithelial integrity. Although ethylene glycol-bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA E3889, Sigma-Aldrich, CABA-Argentina), a calcium chelator, is widely used to disrupt epithelial monolayers in vitro, its application to zebrafish in vivo remains limited. We found that short-term exposure to EGTA induced tail damage ranging from minor and reversible to severe and lethal; at the tissue level, it caused disruption of cadherin-mediated cell-cell contacts and of the barrier function in the epidermis. Labeling with fluorescent quantum dots (QDs) enabled rapid characterization of damage severity, evidencing the disappearance of actin microridges in the enveloping layer. At least 15 min exposure to 25 mM EGTA induces reversible epithelial disruption at the pharyngula stage, providing a chemical model to study healing and regeneration in zebrafish embryos.
期刊介绍:
FEBS Letters is one of the world''s leading journals in molecular biology and is renowned both for its quality of content and speed of production. Bringing together the most important developments in the molecular biosciences, FEBS Letters provides an international forum for Minireviews, Research Letters and Hypotheses that merit urgent publication.