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基于香豆素的非细胞毒性荧光染料,用于在活体成像中追踪肌动蛋白蛋白。

Coumarin-Based Noncytotoxicity Fluorescent Dye for Tracking Actin Protein in In-Vivo Imaging.

机构信息

Neural Developmental Biology Lab, Department of Life Sciences, National Institute of Technology, Rourkela 769008, Odisha, India.

School of Chemistry, Sambalpur University, Jyoti Vihar, Burla, Sambalpur 768019, Odisha, India.

出版信息

Chem Res Toxicol. 2023 Jun 19;36(6):926-933. doi: 10.1021/acs.chemrestox.3c00051. Epub 2023 Jun 1.

Abstract

shares maximum homology with the human disease-causing genes and thus has been employed to evaluate the toxicity of numerous compounds. Further, its distinguishable developmental stages, easy rearing, and short lifespan make it a perfect model organism to study toxicological properties of any new compound. The current study evaluates the toxic effect of a coumarin-based organic fluorescent dye, 7-hydroxy-4-methyl-8-((4-(2-oxo-2-chromen-3-yl)thiazol-2-ylimino)methyl)-2-chromen-2-one (), using as a model organism by studying different behavioral, screening, and staining techniques using flies. For toxicity assessment, one control fly group was compared with various flies that had been subjected to fed dye orally of different concentrations (0.5, 1, 2.5, and 5 μg/mL). The 3rd instar larvae were checked for the larvae crawling assay. The crawling assay demonstrates that the speed and path of the treated larvae are almost equal to the control ones, which signifies the non-neurotoxic property of . Trypan blue assay further suggested that the dye does not cause any major damage to the gut. Phalloidin staining revealed that the actin composition remains unaltered even after the treatment, while the DAPI staining experiment indicates that does not cause any nuclear damage to fly gut cells. However, at a concentration of 5 μg/mL, causes developmental delay. The flies hatched after larval treatment of do not show any structural defects, suggesting clearly that is also nongenotoxic to . The current studies propose as a noncytotoxic and nongenotoxic dye to track actin protein in the model organism .

摘要

与人类疾病相关基因具有最大同源性,因此已被用于评估众多化合物的毒性。此外,其具有明显的发育阶段、易于饲养和短寿命,使其成为研究任何新化合物毒理学特性的理想模式生物。本研究采用 作为模型生物,通过研究不同的行为、筛选和染色技术,评估基于香豆素的有机荧光染料 7-羟基-4-甲基-8-((4-(2-氧代-2-色烯-3-基)噻唑-2-基亚氨基)甲基)-2-色烯-2-酮 ()的毒性。对于毒性评估,将一个对照果蝇组与不同浓度(0.5、1、2.5 和 5μg/mL)的 口服喂养的各种果蝇进行比较。对 3 龄幼虫进行幼虫爬行分析。爬行分析表明,处理幼虫的速度和路径几乎与对照幼虫相同,这表明 没有神经毒性。台盼蓝分析进一步表明,该染料不会对肠道造成任何严重损伤。鬼笔环肽染色显示,即使在 处理后,肌动蛋白组成也没有改变,而 DAPI 染色实验表明, 不会对果蝇肠道细胞造成任何核损伤。然而,在 5μg/mL 浓度下, 会导致发育延迟。经幼虫处理后孵化的果蝇没有表现出任何结构缺陷,这清楚地表明 对 也没有遗传毒性。本研究提出 作为一种非细胞毒性和非遗传毒性染料,用于追踪模型生物中的肌动蛋白蛋白。

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