Subdiffraction-resolution live-cell imaging for visualizing thylakoid membranes.
Subdiffraction-resolution live-cell imaging for visualizing thylakoid membranes.
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DOI：
10.1111/tpj.14021
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发表时间：
2018-10
期刊：
影响因子：
--
通讯作者：
Niyogi KK
中科院分区：
文献类型：
--
作者：
Iwai M;Roth MS;Niyogi KK
叶绿体是含有叶绿素的细胞器，通过光合作用产生能量。在叶绿体内有一个错综复杂的类囊体膜网络，其中含有光合膜蛋白，这些蛋白介导电子传递并产生化学能。从历史上看，电子显微镜一直是可视化类囊体膜的大分子结构和组织的有力工具。然而，由于电子显微镜需要固定和切片，所以了解类囊体膜动力学仍然是难以捉摸的。为了提高对类囊体膜动力学的认识，我们至少需要克服两个问题：1)活体细胞成像条件以可视化体内的活动过程；2)空间分辨率以区分类囊体膜特性。在这里，我们利用三维结构照明显微镜(3D-SIM)来探索研究植物和藻类活细胞中类囊体膜动态的最佳成像条件。我们发现3D-SIM能够检测维管束植物拟南芥叶绿体中类囊体结构的广泛特征，并区分野生型和突变型品系之间的结构差异。利用3D-SIM，我们还显示了绿藻衣藻的整个细胞中的类囊体类组织。这些数据表明，高光强改变了莱茵哈迪尔乳杆菌的类囊体膜结构。此外，为了说明3D-SIM的适用性，我们还观察了绿藻Zofingisis和苔藓Physcomitrella patens。这项研究表明，3D-SIM是研究光自养生物光驯化过程中类囊体膜动态的一种很有前途的方法。
The chloroplast is the chlorophyll-containing organelle that produces energy through photosynthesis. Within the chloroplast is an intricate network of thylakoid membranes containing photosynthetic membrane proteins that mediate electron transport and generate chemical energy. Historically, electron microscopy has been a powerful tool to visualize the macromolecular structure and organization of thylakoid membranes. However, understanding thylakoid membrane dynamics remains elusive because electron microscopy requires fixation and sectioning. To improve knowledge of thylakoid membrane dynamics, we need to overcome at least two issues: 1) the live-cell imaging conditions to visualize active processes in vivo; and 2) the spatial resolution to differentiate thylakoid membrane characteristics. Here, we utilize three-dimensional structured illumination microscopy (3D-SIM) to explore the optimal imaging conditions to investigate the dynamics of thylakoid membranes in live cells of plants and algae. We show that 3D-SIM is capable of examining broad characteristics of thylakoid structures in chloroplasts of the vascular plant Arabidopsis thaliana and distinguishing the structural differences between wild-type and mutant strains. Using 3D-SIM, we also visualize thylakoid organization in whole cells of the green alga Chlamydomonas reinhardtii. These data reveal that high light intensity changes thylakoid membrane structure in C. reinhardtii. Moreover, we observed the green alga Chromochloris zofingiensis and the moss Physcomitrella patens to show the applicability of 3D-SIM. This study demonstrates that 3D-SIM is a promising approach to study the dynamics of thylakoid membranes in photoautotrophic organisms during photoacclimation processes.
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