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Open AccessArticle

3D Dissection of Structural Membrane-Wall Contacts in Filamentous Moss Protonemata

Department of Functional and Evolutionary Ecology, Faculty of Life Sciences, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria
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Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2021, 22(1), 158; https://doi.org/10.3390/ijms22010158
Received: 23 November 2020 / Revised: 19 December 2020 / Accepted: 22 December 2020 / Published: 26 December 2020
(This article belongs to the Special Issue Advances in Plant Cell Imaging)
In conventional light microscopy, the adjacent cell walls of filamentous moss protonemata are seen from its narrow side thereby obscuring the major area of cell–cell connection. Optical sectioning, segmentation and 3D reconstructions allow the tilting and rotation of intracellular structures thereby greatly improving our understanding of interaction between organelles, membranes and the cell wall. Often, the findings also allow for conclusions on the respective functions. The moss Physcomitrium (Physcomitrella) patens is a model organism for growth, development and morphogenesis. Its filamentous protonemata are ideal objects for microscopy. Here, we investigated the cell wall between two neighboring cells and the connection of membranes towards this wall after plasmolysis in 0.8 M mannitol. An m-green fluorescent protein (GFP)-HDEL cell line was used to visualize the endoplasmatic reticulum (ER), the plasma membrane (PM) was stained with FM4-64. Our studies clearly show the importance of cell–cell contacts in P. patens protonemata. In 86% of the investigated cell pairs, at least one of the protoplasts remained fully attached to the adjacent cell wall. By tilting of z-stacks, volume renderings and 3D reconstructions, we visualized the amount of attached/detached PM and ER components after plasmolysis and membrane piercings through the wall of cell neighbors. View Full-Text
Keywords: moss; membrane-wall contact; Physcomitrella sp.; osmotic stress; caulonema; chloronema; 3D imaging moss; membrane-wall contact; Physcomitrella sp.; osmotic stress; caulonema; chloronema; 3D imaging
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    Doi: 10.5281/zenodo.4287438
    Link: https://zenodo.org/record/4287438#.X7vw381KiMo
    Description: Video 1: 3D reconstruction of two adjacent P. patens chloronema cells after plasmolysis (corresponding to Figure 3B-F). The viewer is virtually travelling through the sample in yz-direction. The segmentated 3D model shows the partial detachments (P-P) at the adjoining cell wall. Video 2: Rotating 3D reconstruction of the membrane attachments at the cell wall between two chloronema cells (corresponding to Figure 3B-F). Video 3: 3D reconstruction of two neighboured P. patens caulonema cells after plasmolysis (corresponding to Figure 4). In a virtually generated movie, the sample is observed in yz-direction. We appreciate the high resolution 3D model of a fine ER and membrane network at the wall of two adjacent cells.
MDPI and ACS Style

Harant, D.; Lang, I. 3D Dissection of Structural Membrane-Wall Contacts in Filamentous Moss Protonemata. Int. J. Mol. Sci. 2021, 22, 158.

AMA Style

Harant D, Lang I. 3D Dissection of Structural Membrane-Wall Contacts in Filamentous Moss Protonemata. International Journal of Molecular Sciences. 2021; 22(1):158.

Chicago/Turabian Style

Harant, Dominik; Lang, Ingeborg. 2021. "3D Dissection of Structural Membrane-Wall Contacts in Filamentous Moss Protonemata" Int. J. Mol. Sci. 22, no. 1: 158.

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