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红色 CdSe/ZnS QDs 的细胞内转位及其对酵母极性和肌动蛋白丝的影响。

Red CdSe/ZnS QDs' Intracellular Trafficking and Its Impact on Yeast Polarization and Actin Filament.

机构信息

Department of Biology, Missouri State University, 901 S National, Springfield, MO 65897, USA.

Jordan Valley Innovation Center, 542 N Boonville, Springfield, MO 65806, USA.

出版信息

Cells. 2023 Feb 2;12(3):484. doi: 10.3390/cells12030484.

Abstract

Quantum dots are nanoparticles (2-10 nm) that emit strong and tunable fluorescence. Quantum dots have been heavily used in high-demand commercialized products, research, and for medical purposes. Emerging concerns have demonstrated the negative impact of quantum dots on living cells; however, the intracellular trafficking of QDs in yeast cells and the effect of this interaction remains unclear. The primary goal of our research is to investigate the trafficking path of red cadmium selenide zinc sulfide quantum dots (CdSe/ZnS QDs) in and the impact QDs have on yeast cellular dynamics. Using cells with GFP-tagged reference organelle markers and confocal microscopy, we were able to track the internalization of QDs. We found that QDs initially aggregate at the exterior of yeast cells, enter the cell using clathrin-receptor-mediated endocytosis, and distribute at the late Golgi/trans-Golgi network. We also found that the treatment of red CdSe/ZnS QDs resulted in growth rate reduction and loss of polarized growth in yeast cells. Our RNA sequence analysis revealed many altered genes. Particularly, we found an upregulation of , which has previously been associated with cell cycle arrest when overexpressed, and a downregulation of , a gene that codes for a subunit of AP2 protein important for the recruitment of proteins to clathrin-mediated endocytosis vesicle. Furthermore, CdSe/ZnS QDs treatment resulted in a slightly delayed endocytosis and altered the actin dynamics in yeast cells. We found that QDs caused an increased level of F-actin and a significant reduction in profilin protein expression. In addition, there was a significant elevation in the amount of coronin protein expressed, while the level of cofilin was unchanged. Altogether, this suggests that QDs favor the assembly of actin filaments. Overall, this study provides a novel toxicity mechanism of red CdSe/ZnS QDs on yeast actin dynamics and cellular processes, including endocytosis.

摘要

量子点是纳米颗粒(2-10nm),能发出强且可调的荧光。量子点在需求旺盛的商业化产品、研究和医疗用途中得到了广泛应用。新兴的关注表明,量子点对活细胞有负面影响;然而,量子点在酵母细胞中的细胞内转运以及这种相互作用的影响仍不清楚。我们研究的主要目的是研究红色碲化镉硒锌硫化镉量子点(CdSe/ZnS QDs)在酵母细胞中的转运途径以及 QDs 对酵母细胞动力学的影响。使用带有 GFP 标记的参考细胞器标记物和共焦显微镜的细胞,我们能够追踪 QDs 的内化。我们发现,QDs 最初在酵母细胞的外部聚集,通过网格蛋白受体介导的内吞作用进入细胞,并分布在晚期高尔基体/反式高尔基体网络中。我们还发现,红色 CdSe/ZnS QDs 的处理导致酵母细胞生长速度降低和极化生长丧失。我们的 RNA 序列分析揭示了许多改变的基因。特别是,我们发现上调了基因,当过度表达时,该基因与细胞周期停滞有关,下调了基因,该基因编码 AP2 蛋白的一个亚基,该蛋白对于将蛋白质募集到网格蛋白介导的内吞小泡很重要。此外,CdSe/ZnS QDs 处理导致内吞作用略有延迟,并改变了酵母细胞中的肌动蛋白动力学。我们发现 QDs 导致 F-肌动蛋白水平升高,微丝结合蛋白 Profilin 表达显著降低。此外,冠蛋白的表达量显著增加,而丝切蛋白 Cofilin 的水平不变。总的来说,这表明 QDs 有利于肌动蛋白丝的组装。总的来说,这项研究提供了红色 CdSe/ZnS QDs 对酵母肌动蛋白动力学和细胞过程(包括内吞作用)的新毒性机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a53b/9914768/b0018a106bd6/cells-12-00484-g001.jpg

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