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Recent progress in living cell imaging of plant cytoskeleton and vacuole using fluorescent-protein transgenic lines and three-dimensional imaging

Summary.

In higher-plant cells, microtubules, actin microfilaments, and vacuoles play important roles in a variety of cellular events, including cell division, morphogenesis, and cell differentiation. These intracellular structures undergo dynamic changes in their shapes and functions during cell division and differentiation, and to analyse these sequential structural changes, the vital labelling technique, using the green-fluorescent protein or other fluorescent proteins, has commonly been used to follow the localisation and translocation of specific proteins. To visualise microtubules, actin filaments, and vacuoles, several strategies are available for selecting the appropriate fluorescent-protein fusion partner: microtubule-binding proteins, tubulin, and plus-end-tracking proteins are most suitable for microtubule labelling; the actin binding domain of mouse talin and plant fimbrin for actin microfilament visualisation; and the tonoplast-intrinsic proteins and syntaxin-related proteins for vacuolar imaging. In addition, three-dimensional reconstruction methods are indispensable for localising the widely distributed organelles within the cell. The maximum intensity projection method is suitable for cytoskeletal structures, while contour-based surface modelling possesses many advantages for vacuolar membranes. In this article, we summarise the recent progress in living cell imaging of the plant cytoskeleton and vacuoles using various fusions with green-fluorescent proteins and three-dimensional imaging techniques.

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Abbreviations

3-D:

three-dimensional

ABD:

actin binding domain

AF:

actin microfilament

CLSM:

confocal laser scanning microscopy

CMF:

cellulose microfibril

CMT:

cortical microtubule

GFP:

green-fluorescent protein

LV:

lytic vacuole

MAP:

microtubule-associated protein

MBD:

microtubule binding domain

MIP:

maximum intensity projection

MT:

microtubule

PPB:

preprophase band

PSV:

protein storage vacuole

TIP:

tonoplast-intrinsic protein

+TIP:

plus-end-tracking protein

TVM:

tubular structure of vacuolar membrane

TVS:

transvacuolar strand

VM:

vacuolar membrane

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Correspondence and reprints: Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Kashiwanoha 5-1-5, Kashiwa, Chiba 277-8562, Japan.

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Yoneda, A., Kutsuna, N., Higaki, T. et al. Recent progress in living cell imaging of plant cytoskeleton and vacuole using fluorescent-protein transgenic lines and three-dimensional imaging. Protoplasma 230, 129–139 (2007). https://doi.org/10.1007/s00709-006-0237-4

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  • Keywords: Actin microfilament; Microtubule; Vacuole; Three-dimensional imaging; Green-fluorescent protein.