Abstract
Visualization of biological events in real time in vivo has become a crucial to understand immune responses. We have been established novel visualization tools for life of immune cells: proliferation, cell death, and migration. Fucci-transgenic mice allow us to visualize cell cycle phases by reciprocal expression of mKusabira-Orange2 in G1 phase and mAzami-Green in S/G2/M phase. Caspase-3 indicator SCAT3.1 knock-in mice visualize cell death by changing color. Photoconvertible proteins, Kaede and KikGR expressing mice track cell movement between organs by labeling immune cells as red color. Here, I will introduce how to use and visualize these mice. These techniques will help to understand immune system in the living whole body.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Huang AY, Qi H, Germain RN (2004) Illuminating the landscape of in vivo immunity: insights from dynamic in situ imaging of secondary lymphoid tissues. Immunity 21:331–339
Sumen C, Mempel TR, Mazo IB et al (2004) Intravital microscopy: visualizing immunity in context. Immunity 21:315–329
Tomura M, Sakaue-Sawano A, Mori Y et al (2013) Contrasting quiescent G0 phase with mitotic cell cycling in the mouse immune system. PLoS One 8:e73801
Tomura M, Mori YS, Watanabe R et al (2009) Time-lapse observation of cellular function with fluorescent probe reveals novel CTL-target cell interactions. Int Immunol 21:1145–1150
Tomura M, Yoshida N, Tanaka J et al (2008) Monitoring cellular movement in vivo with photoconvertible fluorescence protein “Kaede” transgenic mice. Proc Natl Acad Sci U S A 105:10871–10876
Tomura M, Hata A, Matsuoka S et al (2014) Tracking and quantification of dendritic cell migration and antigen trafficking between the skin and lymph nodes. Sci Rep 4:6030
Sakaue-Sawano A, Kurokawa H, Morimura T et al (2008) Visualizing spatiotemporal dynamics of multicellular cell-cycle progression. Cell 132:487–498
David R, McIlwain TB, Mak TW (2013) Caspase functions in cell death and disease. Cold Spring Harb Perspect Biol 5:a008656
Takemoto K, Nagai T, Miyawaki A et al (2003) Spatio-temporal activation of caspase revealed by indicator that is insensitive to environmental effects. J Cell Biol 160:235–243
Nagai T, Miyawaki A (2004) A high-throughput method for development of FRET-based indicators for proteolysis. Biochem Biophys Res Commun 319:72–77
Ando R, Hama H, Yamamoto-Hino M et al (2002) An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein. Proc Natl Acad Sci U S A 99:12651–12656
Tsutsui H, Karasawa S, Shimizu H et al (2005) Semi-rational engineering of a coral fluorescent protein into an efficient highlighter. EMBO Rep 6:233–238
Tomura M, Honda T, Tanizaki H et al (2010) Activated regulatory T cells are the major T cell type emigrating from the skin during a cutaneous immune response in mice. J Clin Invest 120:883–893
Tomura M, Itoh K, Kanagawa O (2010) Naive CD4+ T lymphocytes circulate through lymphoid organs to interact with endogenous antigens and upregulate their function. J Immunol 184:4646–4653
Chtanova T, Hampton HR, Waterhouse LA et al (2014) Real-time interactive two-photon photoconversion of recirculating lymphocytes for discontinuous cell tracking in live adult mice. J Biophotonics 7:425–433. 201
Kotani M, Kikuta J, Klauschen F et al (2013) Systemic circulation and bone recruitment of osteoclast precursors tracked by using fluorescent imaging techniques. J Immunol 190:605–612
Shand FH, Ueha S, Otsuji M et al (2014) Tracking of intertissue migration reveals the origins of tumor-infiltrating monocytes. Proc Natl Acad Sci U S A 111:7771–7776
Ikebuchi R, Teraguchi S, Vandenbon A et al (2016) A rare subset of skin-tropic regulatory T cells expressing Il10/Gzmb inhibits the cutaneous immune response. Sci Rep 6:35002
Tomura M, Kabashima K (2013) Analysis of cell movement between skin and other anatomical sites in vivo using photoconvertible fluorescent protein “Kaede”-transgenic mice. Mol Dermatol Methods Mol Biol 961:279–286. Spring (Edited by Cristina Has and Cassian Sitaru)
Acknowledgements
This work was supported in part by JSPS Grants-in-Aid for Scientific Research in Innovative Areas “Analysis and Synthesis of Multidimensional Immune Organ Network” (#24111007); JSPS Grants-in-Aid for Scientific Research (B) (#16H05087); Special Coordination Funds for Promoting Science and Technology of the Japanese Government.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Tomura, M. (2018). New Tools for Imaging of Immune Systems: Visualization of Cell Cycle, Cell Death, and Cell Movement by Using the Mice Lines Expressing Fucci, SCAT3.1, and Kaede and KikGR. In: Ishii, M. (eds) Intravital Imaging of Dynamic Bone and Immune Systems . Methods in Molecular Biology, vol 1763. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7762-8_16
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7762-8_16
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7761-1
Online ISBN: 978-1-4939-7762-8
eBook Packages: Springer Protocols