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Photo-convertible fluorescent proteins as tools for fresh insights on subcellular interactions in plants.
J Microsc. 2016 08; 263(2):148-57.JM

Abstract

Optical highlighters comprise photo-activatable, photo-switchable and photo-convertible fluorescent proteins and are relatively recent additions to the toolbox utilized for live cell imaging research. Here, we provide an overview of four photo-convertible fluorescent proteins (pcFP) that are being used in plant cell research: Eos, Kaede, Maple and Dendra2. Each of these proteins has a significant advantage over other optical highlighters since their green fluorescent nonconverted forms and red fluorescent converted forms are generally clearly visible at expression levels that do not appear to interfere with subcellular dynamics and plant development. These proteins have become increasingly useful for understanding the role of transient and sustained interactions between similar organelles. Tracking of single organelles after green-to-red conversion has provided novel insights on plastids and their stroma-filled extensions and on the formation of mega-mitochondria. Similarly colour recovery after photo-conversion has permitted the estimation of nuclear endo-reduplication events and is being developed further to image protein trafficking within the lumen of the endoplasmic reticulum. We have also applied photo-convertible proteins to create colour-differentiation between similar cell types to follow their development. Both the green and red fluorescent forms of these proteins are compatible with other commonly used single coloured FPs. This has allowed us to develop simultaneous visualization schemes for up to five types of organelles and investigate organelle interactivity. The advantages and caveats associated with the use of photo-convertible fluorescent proteins are discussed.

Authors+Show Affiliations

Laboratory of Plant Development and Interactions, Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada.Laboratory of Plant Development and Interactions, Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada.Laboratory of Plant Development and Interactions, Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada.Laboratory of Plant Development and Interactions, Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada.Laboratory of Plant Development and Interactions, Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada.Laboratory of Plant Development and Interactions, Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada.Laboratory of Plant Development and Interactions, Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Review

Language

eng

PubMed ID

26820914

Citation

Griffiths, N, et al. "Photo-convertible Fluorescent Proteins as Tools for Fresh Insights On Subcellular Interactions in Plants." Journal of Microscopy, vol. 263, no. 2, 2016, pp. 148-57.
Griffiths N, Jaipargas EA, Wozny MR, et al. Photo-convertible fluorescent proteins as tools for fresh insights on subcellular interactions in plants. J Microsc. 2016;263(2):148-57.
Griffiths, N., Jaipargas, E. A., Wozny, M. R., Barton, K. A., Mathur, N., Delfosse, K., & Mathur, J. (2016). Photo-convertible fluorescent proteins as tools for fresh insights on subcellular interactions in plants. Journal of Microscopy, 263(2), 148-57. https://doi.org/10.1111/jmi.12383
Griffiths N, et al. Photo-convertible Fluorescent Proteins as Tools for Fresh Insights On Subcellular Interactions in Plants. J Microsc. 2016;263(2):148-57. PubMed PMID: 26820914.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Photo-convertible fluorescent proteins as tools for fresh insights on subcellular interactions in plants. AU - Griffiths,N, AU - Jaipargas,E-A, AU - Wozny,M R, AU - Barton,K A, AU - Mathur,N, AU - Delfosse,K, AU - Mathur,J, Y1 - 2016/01/28/ PY - 2015/10/31/received PY - 2015/12/22/accepted PY - 2016/1/29/entrez PY - 2016/1/29/pubmed PY - 2018/4/6/medline KW - Dendra2 KW - Kaede KW - mEosFP KW - mMaple KW - organelle interactions KW - photoconvertible fluoresecent proteins SP - 148 EP - 57 JF - Journal of microscopy JO - J Microsc VL - 263 IS - 2 N2 - Optical highlighters comprise photo-activatable, photo-switchable and photo-convertible fluorescent proteins and are relatively recent additions to the toolbox utilized for live cell imaging research. Here, we provide an overview of four photo-convertible fluorescent proteins (pcFP) that are being used in plant cell research: Eos, Kaede, Maple and Dendra2. Each of these proteins has a significant advantage over other optical highlighters since their green fluorescent nonconverted forms and red fluorescent converted forms are generally clearly visible at expression levels that do not appear to interfere with subcellular dynamics and plant development. These proteins have become increasingly useful for understanding the role of transient and sustained interactions between similar organelles. Tracking of single organelles after green-to-red conversion has provided novel insights on plastids and their stroma-filled extensions and on the formation of mega-mitochondria. Similarly colour recovery after photo-conversion has permitted the estimation of nuclear endo-reduplication events and is being developed further to image protein trafficking within the lumen of the endoplasmic reticulum. We have also applied photo-convertible proteins to create colour-differentiation between similar cell types to follow their development. Both the green and red fluorescent forms of these proteins are compatible with other commonly used single coloured FPs. This has allowed us to develop simultaneous visualization schemes for up to five types of organelles and investigate organelle interactivity. The advantages and caveats associated with the use of photo-convertible fluorescent proteins are discussed. SN - 1365-2818 UR - https://www.unboundmedicine.com/medline/citation/26820914/Photo_convertible_fluorescent_proteins_as_tools_for_fresh_insights_on_subcellular_interactions_in_plants_ L2 - https://doi.org/10.1111/jmi.12383 DB - PRIME DP - Unbound Medicine ER -