Abstract
The plant trans-Golgi network (TGN) is a multifunctional organelle derived from the Golgi. It consists of tubulovesicular compartments scattered in the cytosol. They produce secretory vesicles delivering proteins and polysaccharides to the cell wall. They also serve as early endosomal compartments, receiving endocytic cargos from the plasma membrane. This versatility is thought to originate from functional variations among individual TGN compartments. Correlative light and electron microscopy (CLEM) combines the imaging capability of light microscopy and electron microscopy (EM) to determine the location of macromolecules in EM images in the cellular context. It is possible to identify organelles associated with specific fluorescent markers and examine their membrane architectures at nanometer-level resolutions using CLEM. In this chapter, we will explain the CLEM method that our lab uses to investigate functional and structural heterogeneity among individual TGN compartments in plant cells.
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References
Rosquete MR, Drakakaki G (2018) Plant TGN in the stress response: a compartmentalized overview. Curr Opin in Plant Biol 46:122–129
Staehelin LA, Kang B-H (2008) Nanoscale architecture of endoplasmic reticulum export sites and of Golgi membranes as determined by electron tomography. Plant Physiol 147:1454–1468
van de Meene AML, Doblin MS, Bacic A (2016) The plant secretory pathway seen through the lens of the cell wall. Protoplasma 254:1–20
LaMontagne ED, Heese A (2017) Trans -Golgi network/early endosome: a central sorting station for cargo proteins in plant immunity. Curr Opin in Plant Biol 40:114–121
Ruiz Rosquete M, Davis DJ, Drakakaki G (2017) The plant Trans-Golgi network. Not just a matter of distinction. Plant Physiol 176:01239.2017–01239.2198
Dettmer J, Hong-Hermesdorf A, Stierhof Y-D, Schumacher K (2006) Vacuolar H+-ATPase activity is required for endocytic and secretory trafficking in Arabidopsis. Plant Cell 18:715–730
Luschnig C, Vert G (2014) The dynamics of plant plasma membrane proteins: PINs and beyond. Development 141:2924–2938
Kang B-H, Nielsen E, Preuss ML, Mastronarde D, Staehelin LA (2011) Electron tomography of RabA4b- and PI-4Kβ1-labeled trans golgi network compartments in Arabidopsis. Traffic 12:313–329
Uemura T, Nakano RT, Takagi J, Wang Y, Kramer K, Finkemeier I, Nakagami H, Tsuda K, Ueda T, Schulze-Lefert P, Nakano A (2019) A Golgi-released subpopulation of the trans-Golgi network mediates protein secretion in Arabidopsis. Plant Physiol 179:519–532
Wang P, Chen X, Goldbeck C, Chung E, Kang B-H (2017) A distinct class of vesicles derived from the trans-Golgi mediates secretion of xylogalacturonan in the root border cell. Plant J 92:596–610
Wang P, Liang Z, Kang B-H (2019) Electron tomography of plant organelles and the outlook for correlative microscopic approaches. New Phytol 223:1756–1761
SeguÃ-Simarro JM, Staehelin LA (2006) Cell cycle-dependent changes in Golgi stacks, vacuoles, clathrin-coated vesicles and multivesicular bodies in meristematic cells of Arabidopsis thaliana: a quantitative and spatial analysis. Planta 223:223–236
Kukulski W, Schorb M, Welsch S, Picco A, Kaksonen M, Briggs JAG (2011) Correlated fluorescence and 3D electron microscopy with high sensitivity and spatial precision. J Cell Biol 192:111–119
Bell K, Mitchell S, Paultre D, Posch M, Oparka K (2013) Correlative imaging of fluorescent proteins in resin-embedded plant material. Plant Physiol 161:1595–1603
Avinoam O, Schorb M, Beese CJ, Briggs JAG, Kaksonen M (2015) ENDOCYTOSIS. Endocytic sites mature by continuous bending and remodeling of the clathrin coat. Science 348:1369–1372
Zachari M, Gudmundsson SR, Li Z, Manifava M, Shah R, Smith M, Stronge J, Karanasios E, Piunti C, Kishi-Itakura C, Vihinen H, Jokitalo E, Guan J-L, Buss F, Smith AM, Walker SA, Eskelinen E-L, Ktistakis NT (2019) Selective autophagy of mitochondria on a ubiquitin- endoplasmic-reticulum platform. Dev Cell 50(5):627–643
Kang B (2010) Electron microscopy and high-pressure freezing of Arabidopsis. Methods Cell Biol 96:259–283
Wang P, Kang B-H (2018) The trans-Golgi sorting and the exocytosis of xylogalacturonan from the root border/border-like cell are conserved among monocot and dicot plant species. Plant Signal Behav 13:e1469362–e1469363
Toyooka K, Kang B-H (2014) Reconstructing plant cells in 3D by serial section electron tomography. In: Zarsky V, Cvrckova F (eds) Plant cell morphogenesis. Humana Press, Totowa, NJ, pp 159–170
Kang B-H (2016) STEM tomography imaging of hypertrophied Golgi stacks in mucilage-secreting cells. Methods Mol Biol 1496:55–62
Mai KKK, Kang B-H (2017) Semiautomatic segmentation of plant Golgi stacks in electron tomograms using 3dmod. Methods Mol Biol 1662:97–104
Liang Z, Zhu N, Mai KK, Liu Z, Liu Z, Tzeng D, Tzeng DTW, Osteryoung KW, Zhong S, Staehelin LA, Staehelin A, Kang B-H (2018) Thylakoid-bound polysomes and a dynamin-related protein, FZL, mediate critical stages of the linear chloroplast biogenesis program in greening Arabidopsis cotyledons. Plant Cell 30:1476–1495
Acknowledgments
This work was supported by Hong Kong Research Grant Council (GRF14126116, AoE/M-05/12, C4002-17G), Rural Development Administration of Korea (Project No. 10953092019), and Chinese University of Hong Kong (Direct Grant 14101218).
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Wang, P., Kang, BH. (2020). Correlative Light and Electron Microscopy Imaging of the Plant trans-Golgi Network. In: Otegui, M. (eds) Plant Endosomes. Methods in Molecular Biology, vol 2177. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0767-1_6
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DOI: https://doi.org/10.1007/978-1-0716-0767-1_6
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