使用活细胞成像探索球石藻中的细胞内离子池,Advanced Biosystems

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使用活细胞成像探索球石藻中的细胞内离子池
Advanced Biosystems Pub Date : 2021-04-14 , DOI: 10.1002/adbi.202000296
Hadas Peled-Zehavi ₁ , Assaf Gal ₁

Affiliation

一些微生物，例如球石藻，会产生由无机固体制成的复杂外骨骼。球石，即球石藻的碳酸钙鳞片，是这种复杂 3D 结构精确生物生产的例子。然而，对控制细胞内矿物质形成的细胞机制，特别是这些微藻运输高通量无机构建块的能力的理解仍然有限。最近，使用冷冻电子和 X 射线显微镜，显示几个细胞内隔室储存高浓度的钙和磷，并被认为在细胞内矿化途径中起主导作用。在这里，活细胞共聚焦显微镜和荧光标记用于检查球石藻中离子储存的动态。使用钙黄绿素和 4'，6-二脒基-2-苯基吲哚（DAPI）作为钙和聚磷酸盐的荧光代理，这些实验揭示了在各种菌株和条件下具有不同荧光特征的意外过多的细胞器。令人惊讶的是，荧光标记并未显示钙化过程中的变化，并且在钙化细胞和非钙化细胞之间是相似的，这表明这些离子池可能不是钙转运的动态途径。在这种情况下，球石藻维持细胞内钙化能力背后的谜团仍有待全面阐明。荧光标记不显示钙化过程中的变化，并且在钙化和非钙化细胞之间是相似的，这表明这些离子库可能不是钙运输的动态途径。在这种情况下，球石藻维持细胞内钙化能力背后的谜团仍有待全面阐明。荧光标记不显示钙化过程中的变化，并且在钙化细胞和非钙化细胞之间是相似的，这表明这些离子池可能不是钙转运的动态途径。在这种情况下，球石藻维持细胞内钙化能力背后的谜团仍有待全面阐明。

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Exploring Intracellular Ion Pools in Coccolithophores Using Live-Cell Imaging

Some microorganisms, such as coccolithophores, produce an intricate exoskeleton made of inorganic solids. Coccoliths, the calcium carbonate scales of coccolithophores, are examples of the precise bioproduction of such complex 3D structures. However, the understanding of the cellular mechanisms that control mineral formation inside the cell, specifically the ability of these microalgae to transport high fluxes of inorganic building blocks, is still limited. Recently, using cryo-electron and X-ray microscopy, several intracellular compartments are shown to store high concentrations of calcium and phosphorous and are suggested to have a dominant role in the intracellular mineralization pathway. Here, live-cell confocal microscopy and fluorescent markers are used to examine the dynamics of ion stores in coccolithophores. Using calcein and 4′,6-diamidino-2-phenylindole (DAPI) as fluorescent proxies for calcium and polyphosphates, the experiments reveal an unexpected plethora of organelles with distinct fluorescent signatures over a wide range of strains and conditions. Surprisingly, the fluorescent labeling does not show changes along the calcification process and is similar between calcifying and noncalcifying cells, suggesting that these ion pools may not be a dynamic avenue for calcium transport. In such a case, the enigma behind the ability of coccolithophores to sustain intracellular calcification still awaits comprehensive elucidation.

更新日期：2021-06-17

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