Abstract

Damage to the plasma membrane (PM) of cells in pea leaf epidermis determined by its permeability to propidium iodide (PI), which binds to DNA of cell nuclei, and programmed cell death (PCD) detected by the destruction of cell nuclei were investigated. PM of the epidermal cells in the isolated epidermis was permeable to PI (it stained their nuclei). PM of the guard cells did not allow PI to pass through. KCN, an inducer of PCD, caused the destruction of both epidermal and guard cell nuclei. KCN-induced destruction of guard cell nuclei was accompanied by the penetration of PI into the cells. The polycation chitosan at a concentration of 0.1 mg/mL caused the destruction of the epidermal cell nuclei, but the permeability of the guard cell PM for PI staining their nuclei was induced at a concentration of 1 mg/mL. Other polycations (cytochrome c, polylysine, polyethylenimine, and protamine) also caused staining of the guard cell nuclei by PI. Polyanions (polyacrylic acid, dextran, and heparin) initiated the destruction of cell nuclei, which was accompanied by the penetration of PI into cells. Detergents Triton X-100 and lauryldimethylamine-N-oxide produced the permeability of the guard cell PM for PI and prevented the destruction of the nuclei that was induced by KCN. Treatment of the epidermis with Triton X-100 (for 2 h with its subsequent washing) increased the destruction of the guard cell nuclei that was caused by KCN. Polycations polyethyleneimine and protamine were prevented, while chitosan, cytochrome c, and polylysine, on the contrary, enhanced KCN-induced destruction of the guard cell nuclei. The obtained data shows that the destruction of cell nuclei upon induction of cell death with KCN or polyanions is accompanied by damage to PM (producing its permeability for PI). Damage to PM caused by detergents or polycations prior to cell treatment by KCN can prevent or, on the contrary, intensify the destruction of cell nuclei.

中文翻译：

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碘化丙啶血浆膜的渗透性和豌豆叶表皮中细胞核的破坏：聚电解质和洗涤剂的作用

摘要

研究了豌豆叶表皮中细胞质膜（PM）的损伤，该损伤是通过与细胞核DNA结合的碘化丙啶（PI）的渗透性确定的，并通过破坏细胞核来检测程序性细胞死亡（PCD）。分离的表皮中的表皮细胞的PM对PI是可渗透的（染色了它们的细胞核）。保卫室的PM不允许PI通过。PCD的诱导剂KCN引起表皮和保卫细胞核的破坏。KCN诱导的保卫细胞核破坏伴随着PI渗透进入细胞。浓度为0.1 mg / mL的聚阳离子壳聚糖引起表皮细胞核的破坏，但浓度为1 mg / mL诱导保卫细胞PM对PI染色其核的渗透性。其他聚阳离子（细胞色素c，聚赖氨酸，聚乙烯亚胺和鱼精蛋白）也引起PI对保卫细胞核的染色。聚阴离子（聚丙烯酸，右旋糖酐和肝素）引发细胞核的破坏，并伴随着PI渗透进入细胞。清洁剂Triton X-100和月桂基二甲基胺-N-氧化物可产生保卫细胞PM对PI的渗透性，并防止KCN诱导的细胞核破坏。用Triton X-100处理表皮（持续2小时，随后清洗）增加了由KCN引起的保卫细胞核的破坏。防止了聚阳离子聚乙烯亚胺和鱼精蛋白，而壳聚糖，细胞色素c相反，多聚赖氨酸和多赖氨酸增强了KCN诱导的保卫细胞核破坏。获得的数据表明，用KCN或聚阴离子诱导细胞死亡后破坏细胞核会伴随PM的破坏（产生其对PI的渗透性）。在用KCN进行细胞处理之前，清洁剂或聚阳离子对PM造成的损害可以阻止或相反地加剧细胞核的破坏。