Russian Journal of Biological Physics and Chemisrty Russian Journal of Biological Physics and Chemisrty АКТУАЛЬНЫЕ ВОПРОСЫ БИОЛОГИЧЕСКОЙ ФИЗИКИ И ХИМИИ 2499-9962 54312 Общая биофизика General biophysics Общая биофизика INTRACELLULAR TRANSPORT IN MOUSE OOCYTES DURING FEMTOSECOND LASER CELL FUSION ВНУТРИКЛЕТОЧНЫЙ ТРАНСПОРТ В ООЦИТАХ МЫШИ ПРИ СЛИЯНИИ ФЕМТОСЕКУНДНЫМИ ЛАЗЕРНЫМИ ИМПУЛЬСАМИ Шахов А М Shakhov A M physics2007@yandex.ru Осыченко А А Osychenko A A Астафьев А А Astafiev A A Серобян Г А Serobyan G A Надточенко В А Nadtochenko V A Институт химической физики им. Н.Н. Семенова (ИХФ РАН) ru Semenov Institute of Chemical Physics (ICP RAS) ru Институт химической физики им. Н.Н. Семенова (ИХФ РАН) ru Semenov Institute of Chemical Physics (ICP RAS) ru Институт химической физики им. Н.Н. Семенова (ИХФ РАН) ru Semenov Institute of Chemical Physics (ICP RAS) ru Институт химической физики им. Н.Н. Семенова (ИХФ РАН) ru Semenov Institute of Chemical Physics (ICP RAS) ru Институт химической физики им. Н.Н. Семенова (ИХФ РАН) ru Semenov Institute of Chemical Physics (ICP RAS) ru 25 09 2018 25 09 2018 3 3 467 471 20 09 2018 20 09 2018 https://rusjbpc.ru/en/nauka/article/54312/view

В работе представлены результаты по отслеживанию внутриклеточного транспорта в клетках млекопитающих во время искусственного слияния. В качестве модельного объекта были использованы ооциты мыши, которые подвергались процедуре лазерного слияния. Воздействие фемтосекундными лазерными импульсами в месте контакта двух ооцитов приводило к перфорации и объединению мембран и медленному изменению формы слитого ооцита с эллипсоидной на сферическую. В результате слияния происходил сложный процесс перемешивания цитоплазмы исходных ооцитов, задействующий механизмы внутриклеточного транспорта и активной диффузии. Для визуализации данных механизмов были применены методы флуоресцентной микроскопии основанные на GFP и BODIPY. Установлено, что мелкие частицы в цитозоле ооцитов перемешивается преимущественно диффузионно, в то время как органеллы и цитоскелет, не способные к диффузионному транспорту, не перемешиваются после окончания слияния в течении нескольких часов.

Here we demonstrate intracellular transport in fused mammalian cells. Mouse oocytes as a model system were subjected to the laser cell fusion. The contact point of the two oocytes cytoplasmic membranes was exposed to femtosecond laser pulses, which produce the membrane perforation and fusion and a slow change in the shape of the fused oocyte from ellipsoidal to spherical. A complex process of original oocytes cytoplasm mixing took place as a result of the fusion, involving mechanisms of intracellular transport and active diffusion. Fluorescence microscopy methods based on GFP and BODIPY staining were applied to visualize these mechanisms. We revealed that the small particles in cytosol of the oocytes are mixed almost diffusively, while the organelles and cytoskeleton, that are incapable of diffusion, are not mixed even within several hours after the very end of the oocyte fusion.

 ооциты мыши лазерное слияние клеток фемтосекундные лазеры GFP BODIPY mouse oocytes laser cell fusion femtosecond laser

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