Live-Cell Imaging of Dual-Labeled Golgi Stacks in Tobacco BY-2 Cells Reveals Similar Behaviors for Different Cisternae during Movement and Brefeldin A Treatment

作     者：Stephanie L. Madison Andreas Nebenfuhr 

作者机构：Department of Biochemistry and Cellular and Molecular Biology University of Tennessee Knoxville TN 37996-0840 USA 

出 版 物：《Molecular Plant》 (分子植物（英文版）)

年 卷 期：2011年第4卷第5期

页      面：896-908页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 08[工学] 071009[理学-细胞生物学] 0802[工学-机械工程] 0901[农学-作物学] 0902[农学-园艺学] 

基　　金：National Science Foundation, NSF: MCB-0822111 Directorate for Biological Sciences, BIO: 0822111 

主　　题：Golgi apparatus Golgi stack integrity brefeldin A monensin tobacco BY-2 cells live-cell imaging. 

摘      要：In plant cells, the Golgi apparatus consists of numerous stacks that, in turn, are composed of several flattened cisternae with a clear cis-to-trans polarity. During normal functioning within living cells, this unusual organelle displays a wide range of dynamic behaviors such as whole stack motility, constant membrane flux through the cisternae, and Golgi enzyme recycling through the ER. In order to further investigate various aspects of Golgi stack dynamics and integrity, we co-expressed pairs of established Golgi markers in tobacco BY-2 cells to distinguish sub-compartments of the Golgi during monensin treatments, movement, and brefeldin A （BFA）-induced disassembly. A combination of cis and trans markers revealed that Golgi stacks remain intact as they move through the cytoplasm. The Golgi stack orientation during these movements showed a slight preference for the cis side moving ahead, but trans cisternae were also found at the leading edge. During BFA treatments, the different sub-compartments of about half of the observed stacks fused with the ER sequentially; however, no consistent order could be detected. In contrast, the ionophore monensin resulted in swelling of trans cisternae while medial and particularly cis cisternae were mostly unaffected. Our results thus demonstrate a re- markable equivalence of the different cisternae with respect to movement and BFA-induced fusion with the ER. In addi- tion, we propose that a combination of dual-label fluorescence microscopy and drug treatments can provide a simple alternative approach to the determination of protein localization to specific Golgi sub-compartments.