Russian Journal of Biological Physics and Chemisrty Russian Journal of Biological Physics and Chemisrty АКТУАЛЬНЫЕ ВОПРОСЫ БИОЛОГИЧЕСКОЙ ФИЗИКИ И ХИМИИ 2499-9962 54314 Общая биофизика General biophysics Общая биофизика AGONIST INDUCED CA2+ SIGNALING IN MESENCHYMAL STROMAL CELLS АГОНИСТ-ИНДУЦИРУЕМАЯ СА2+ СИГНАЛИЗАЦИЯ В МЕЗЕНХИМНЫХ СТРОМАЛЬНЫХ КЛЕТКАХ Котова П Д Kotova P D polinakotova88@gmail.com Ивашин Д С Ivashin D S Колесников С С Kolesnikov S S Институт биофизики клетки РАН ru Institute of Cell Biophysics, Russian Academy of Sciences ru Институт биофизики клетки РАН ru Institute of Cell Biophysics, Russian Academy of Sciences ru Институт биофизики клетки РАН ru Institute of Cell Biophysics, Russian Academy of Sciences ru 25 09 2018 25 09 2018 3 3 476 483 20 09 2018 20 09 2018 https://rusjbpc.ru/en/nauka/article/54314/view

Мезенхимные стромальные клетки (МСК) из различных источников представляют собой гетерогенную популяцию недифференцированных пролиферирующих клеток, которая содержит мультипотентные стволовые клетки, способные к дифференцировке в клетки нескольких мезенхимных линий. Используя микрофотометрию (Ca2+ imaging) и Ca2+-чувствительный флуорофор Fluo-4, мы исследовали Ca2+ сигналы, инициируемые агонистами ряда GPCR рецепторов в МСК из жировой ткани человека. Интересной особенностью агонист-индуцированной Ca2+ сигнализации в МСК было то, что кратковременная стимуляция клеток вызывала Ca2+ ответы, которые генерировались по принципу “все-или-ничего”. Более точно, при относительно низких дозах агонисты не вызывали детектируемого изменения внутриклеточного Са2+, но стимулировали большие Са2+ ответы, которые были примерно одинаковы по амплитуде при различных концентрациях агонистов, превышающих пороговую. Последняя была оценены как 0.1, 0.5, 1 и 2 мкM для норадреналина, ADP, ATP, и UTP, соответственно. Результаты экспериментов с использованием ингибиторного анализа и фотолиза химических групп (uncaging) свидетельствовали о том, что фосфоинозитидный каскад и Ca2+ индуцированный выброс депонированного Ca2+ (Ca2+-induced Ca2+ release (CICR)) вовлечены в формирование агонист-зависимых Ca2+ сигналов в цитоплазме МСК. В целом, полученные данные позволяют рассматривать трансдукцию исследовавшихся агонистов как двух-стадийный процесс. Первоначально агонист стимулирует локальный Ca2+ сигнал, который, скорее всего, градуально зависит от его дозы. При превышении порога этот локальный Ca2+ сигнал стимулирует CICR, который конвертирует его в глобальный Ca2+ сигнал и придает окончательную форму Ca2+ ответу на агонист, делая его универсальным при различных концентрациях, превышающих пороговую.

Mesenchymal stromal cells (MSCs) from different sources represent a heterogeneous population of proliferating non-differentiated cells that contains multipotent stem cells capable of originating a variety of mesenchymal cell lineages. By using Ca2+ imaging and the Ca2+ dye Fluo-4, we studied MSCs from the human adipose tissue and examined Ca2+ signaling initiated by purinergic agonists. Although all tested compounds were capable of mobilizing intracellular Ca2+ in MSCs, sensitivity of individual MSCs to a particular agonist varied from cell to cell. Being characterized by a relative change of Fluo-4 fluorescence, agonist-induced Ca2+ responses were generated in an “all-or-nothing” fashion. Specifically, at relatively low doses, agonists elicited undetectable responses but initiated quite similar Ca2+ transients of large magnitude at all concentrations above the threshold, which was nearly 0.1, 0.2, 0.5, 1, and 2 µM for noradrenaline, ADP, ATP, and UTP, respectively. The inhibitory analysis and Ca2+/IP3 uncaging pointed at the phosphoinositide cascade as a pivotal pathway responsible for agonist transduction and implicated Ca2+-induced Ca2+ release (CICR) mediated by IP3 receptors in shaping agonists-dependent Ca2+ signals. Altogether, our data suggest that agonist transduction in MSCs includes two fundamentally different stages: an agonist initially triggers a local, gradual, and relatively small Ca2+ signal, which next stimulates CICR to accomplish transduction with a large and global Ca2+ transient. By involving the trigger-like mechanism CICR, a cell is capable of generating Ca2+ responses of virtually universal shape and magnitude at different agonist concentrations above the threshold.

 Ca2+ сигнализация гептаспиральные рецепторы calcium-induced calcium release IP3 рецепторы мезенхимные стромальные клетки Ca2+ signaling G-protein coupled receptors calcium-induced calcium release IP3 receptors mesenchymal stromal cells

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