Influence of the cell wall on intracellular delivery to algal cells by electroporation and sonication
Influence of the cell wall on intracellular delivery to algal cells by electroporation and sonication
复制标题
DOI：
10.1016/j.ultrasmedbio.2007.05.008
复制
发表时间：
2007-11-01
影响因子：
2.9
通讯作者：
Prausnitz, Mark R.
中科院分区：
文献类型：
--
作者：
Azencott, Harold R.;Peter, Gary F.;Prausnitz, Mark R.
为了评估细胞壁作为细胞内转运屏障的作用，野生型莱茵衣藻细胞和没有细胞壁的突变细胞被暴露于电穿孔或超声波处理。流式细胞仪检测细胞内钙黄绿素和牛血清白蛋白(BSA)摄取和细胞活力损失作为电穿孔跨膜电位和声能的函数。野生型细胞的电穿孔随着跨膜电位的增加而增加钙蛋白的摄取，但提供的牛血清白蛋白要少得多。壁缺陷细胞的电穿孔对钙黄绿素的摄取也有类似的影响，但与野生型细胞相比，BSA的摄取增加了7.5倍，这表明在电穿孔过程中，细胞壁是BSA输送的重要屏障。对野生型细胞进行超声波处理后，钙黄绿素和牛血清白蛋白的摄取水平相似。这表明BSA传递的细胞壁屏障可以通过超声波来克服。电穿孔跨膜电位或声能的增加也会导致细胞活力损失的增加，在这些细胞中，壁缺陷的细胞特别容易溶解。总体而言，我们认为这是第一个直接比较电穿孔和超声波在任何细胞类型中的效果的研究。具体地说，这些发现表明，电穿孔主要通过质膜运输分子，因为它的机制特定于脂质双层破坏，而超声波运输分子跨越质膜和细胞壁，因为它非特异性地破坏细胞表面屏障。(电子邮件：Prausnitz@gatech.edu)(C)2007年世界医学和生物学超声联合会。
To assess the cell wall's role as a barrier to intracellular delivery, wild-type Chlaynydomonas reinhardtii algal cells and mutant cells lacking a cell wall were exposed to electroporation or sonication. Flow cytometry determined intracellular uptake of calcein and bovine serum albumin (BSA) and loss of cell viability as functions of electroporation transmembrane potential and acoustic energy. Electroporation of wild-type cells increased calcein uptake with increasing transmembrane potential, but delivered much less BSA. Electroporation of wall-deficient cells had similar effects on calcein uptake, but increased BSA uptake as much as 7.5-fold relative to wild-type cells, which indicated that the cell wall was a significant barrier to BSA delivery during electroporation. Sonication of wild-type cells caused calcein and BSA uptake at similar levels. This suggests that the cell wall barrier to BSA delivery can be overcome by sonication. Increased electroporation transmembrane potential or acoustic energy also caused increased loss of cell viability, where wall-deficient cells were especially susceptible to lysis. Overall, we believe this is the first study to compare directly the effects of electroporation and sonication in any cell type. Specifically, these findings suggest that electroporation primarily transports molecules across the plasma membrane because its mechanism is specific to lipid bilayer disruption, whereas sonication transports molecules across both the plasma membrane and cell wall, because it nonspecifically disrupts cell-surface barriers. (E-mail: prausnitz@gatech.edu) (c) 2007 World Federation for Ultrasound in Medicine & Biology.