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Electron tomographic characterization of a vacuolar reticulum and of six vesicle types that occupy different cytoplasmic domains in the apex of tip-growing Chara rhizoids

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Abstract

We provide a 3D ultrastructural analysis of the membrane systems involved in tip growth of rhizoids of the green alga Chara. Electron tomography of cells preserved by high-pressure freeze fixation has enabled us to distinguish six different types of vesicles in the apical cytoplasm where the tip growth machinery is accommodated. The vesicle types are: dark and light secretory vesicles, plasma membrane-associated clathrin-coated vesicles (PM-CCVs), Spitzenkoerper-associated clathrin-coated vesicles (Sp-CCVs) and coated vesicles (Sp-CVs), and microvesicles. Each of these vesicle types exhibits a distinct distribution pattern, which provides insights into their possible function for tip growth. The PM-CCVs are confined to the cytoplasm adjacent to the apical plasma membrane. Within this space they are arranged in clusters often surrounding tubular plasma membrane invaginations from which CCVs bud. This suggests that endocytosis and membrane recycling are locally confined to specialized apical endocytosis sites. In contrast, exocytosis of secretory vesicles occurs over the entire membrane area of the apical dome. The Sp-CCVs and the Sp-CVs are associated with the aggregate of endoplasmic reticulum membranes in the center of the growth-organizing Spitzenkoerper complex. Here, Sp-CCVs are seen to bud from undefined tubular membranes. The subapical region of rhizoids contains a vacuolar reticulum that extends along the longitudinal cell axis and consists of large, vesicle-like segments interconnected by thin tubular domains. The tubular domains are encompassed by thin filamentous structures resembling dynamin spirals which could drive peristaltic movements of the vacuolar reticulum similar to those observed in fungal hyphae. The vacuolar reticulum appears to serve as a lytic compartment into which multivesicular bodies deliver their internal vesicles for molecular recycling and degradation.

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Abbreviations

D-SV:

Dark secretory vesicle

ER:

Endoplasmic reticulum

L-SV:

Light secretory vesicle

MV:

Microvesicle

PM:

Plasma membrane

PM-CCV:

PM-associated clathrin-coated vesicle

Sp-CCV:

Spitzenkoerper-associated CCV

Sp-CV:

Spitzenkoerper-associated coated vesicle

SV:

Secretory vesicle

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Acknowledgments

We are grateful to Zachary Gergely and Byung-Ho Kang (MCDB, University of Colorado, Boulder, CO, USA) for their valuable help. Many thanks are also due to the members of the Boulder Laboratory for 3D Electron Microscopy of Cells (MCDB, University of Colorado, Boulder, CO, USA, Grant RR00592). Brigitte Buchen (University of Bonn, Germany) is gratefully acknowledged for helpful comments and critical reading of the manuscript. This work was supported by the Deutsches Zentrum für Luft- und Raumfahrt (DLR) on behalf of the Bundesministerium für Wirtschaft und Technologie (grant 50WB0515), by National Institute of Health Grant GM 61306 to L.A.S., and by a fellowship of the Cusanuswerk to C.L.

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Authors and Affiliations

  1. Gravitationsbiologie, Institut für Molekulare Physiologie und Biotechnologie der Pflanzen, Universität Bonn, Kirschallee 1, 53115, Bonn, Germany

    Christoph Limbach, Andreas Sievers & Markus Braun

  2. Department of Molecular, Cellular and Developmental Biology, University of Colorado, UCB 347, Boulder, CO, 80309-0347, USA

    L. Andrew Staehelin

  3. Department of Cell and Molecular Biology, Biomedical Center, Uppsala University, Box 596, 75124, Uppsala, Sweden

    Christoph Limbach

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  1. Christoph Limbach
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  2. L. Andrew Staehelin
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Correspondence to Christoph Limbach.

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Limbach, C., Staehelin, L.A., Sievers, A. et al. Electron tomographic characterization of a vacuolar reticulum and of six vesicle types that occupy different cytoplasmic domains in the apex of tip-growing Chara rhizoids. Planta 227, 1101–1114 (2008). https://doi.org/10.1007/s00425-007-0684-y

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