vestnik-bio-msuВестник Московского университета. Серия 16. БиологияVestnik Moskovskogo universiteta. Seriya 16. Biologiya0137-0952Lomonosov Moscow State University, School of Biologyvestnik-bio-msu-767Research ArticleОРИГИНАЛЬНОЕ ИССЛЕДОВАНИЕRESEARCH ARTICLEПроницаемость плазматической мембраны для йодида пропидия и разрушение ядер клеток в эпидермисе листьев гороха: действие полиэлектролитов и детергентовPermeability of the plasma membrane for propidium iodide and destruction of cell nuclei in the epidermis of pea leaves: the effect of polyelectrolytes and detergentsКиселевскийД. Б.KiselevskyD. B.

Киселевский Дмитрий Борисович – канд. биол. наук, ст. науч. сотр. кафедры иммунологии биологического факультета

119234, г. Москва, Ленинские горы, д. 1, стр. 12

Department of Immunology, School of Biology

Leninskiye gory 1–12, Moscow, 119234

dkiselevs@mail.ruСамуиловВ. Д.SamuilovV. D.

Самуилов Виталий Дмитриевич – докт. биол. наук, проф., гл. науч. сотр. кафедры иммунологии биологического факультета

119234, г. Москва, Ленинские горы, д. 1, стр. 12

Department of Immunology, School of Biology

Leninskiye gory 1–12, Moscow, 119234

Московский государственный университет имени М.В. ЛомоносоваРоссияLomonosov Moscow State UniversityRussian Federation201921092019743188194Copyright © Киселевский Д.Б., Самуилов В.Д., 20192019Киселевский Д.Б., Самуилов В.Д.Kiselevsky D.B., Samuilov V.D.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vestnik-bio-msu.elpub.ru/jour/article/view/767

Исследовали повреждение плазматической мембраны (ПМ) клеток в эпидермисе листьев гороха, определяемое по ее проницаемости для йодида пропидия (propidium iodide, PI), связывающегося с ДНК клеточных ядер, и программируемую клеточную смерть, регистрируемую по разрушению клеточных ядер. ПМ эпидермальных клеток в изолированном эпидермисе проницаема для PI (он окрашивал их ядра). ПМ устьичных клеток не пропускала PI. Индуктор программируемой клеточной смерти KCN вызывал разрушение ядер и эпидермальных, и устьичных клеток. KCN-индуцированный распад ядер устьичных клеток сопровождался проникновением PI в клетки. Поликатион хитозан в концентрации 0,1 мг/мл вызывал разрушение ядер эпидермальных клеток, а в концентрации 1 мг/мл приводил к возникновению проницаемости ПМ устьичных клеток для PI, окрашивающего их ядра. Другие поликатионы (цитохром с, полилизин, полиэтиленимин и протамин) тоже вызывали окрашивание ядер устьичных клеток PI. Полианионы (полиакриловая кислота, декстран и гепарин) инициировали разрушение клеточных ядер, которому сопутствовало проникновение PI в клетки. Детергенты тритон Х-100 и лаурилдиметиламинN-оксид индуцировали проницаемость ПМ устьичных клеток для PI. KCN не вызывал разрушение ядер устьичных клеток в эпидермисе, обработанном детергентами. Обработка эпидермиса тритоном Х-100 (в течение 2 ч с его последующей отмывкой) усиливала разрушение ядер устьичных клеток, вызванное KCN. Поликатионы полиэтиленимин и протамин предотвращали, а хитозан, цитохром с и полилизин, напротив, усиливали KCN-индуцированное разрушение ядер устьичных клеток. Результаты свидетельствуют о том, что разрушение клеточных ядер при индукции гибели клеток KCN или полианионами сопровождается повреждением ПМ (создающим ее проницаемость для PI). Повреждение ПМ, вызванное детергентами или поликатионами до обработки клеток KCN, может предотвращать или, напротив, усиливать разрушение клеточных ядер.

 

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 was 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 the 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 at a concentration of 1 mg/ ml, the permeability of the guard cell PM for PI staining their nuclei was induced. 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 data obtained 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 treatm       by KCN, can prevent or, on the contrary, intensify the destruction of cell nuclei.

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The authors declare that there are no conflicts of interest present.