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Improved clearing method contributes to deep imaging of plant organs

Sakamoto, Yuki Ishimoto, Anna Sakai, Yuuki Sato, Moeko Nishihama, Ryuichi Abe, Konami Sano, Yoshitake Furuichi, Teiichi Tsuji, Hiroyuki Kohchi, Takayuki Matsunaga, Sachihiro 神戸大学

2022.01.10

概要

Tissue clearing methods are increasingly essential for the microscopic observation of internal tissues of thick biological organs. We previously developed TOMEI, a clearing method for plant tissues; however, it could not entirely remove chlorophylls nor reduce the fluorescent signal of fluorescent proteins. Here, we developed an improved TOMEI method (iTOMEI) to overcome these limitations. First, a caprylyl sulfobetaine was determined to efficiently remove chlorophylls from Arabidopsis thaliana seedlings without GFP quenching. Next, a weak alkaline solution restored GFP fluorescence, which was mainly lost during fixation, and an iohexol solution with a high refractive index increased sample transparency. These procedures were integrated to form iTOMEI. iTOMEI enables the detection of much brighter fluorescence than previous methods in tissues of A. thaliana, Oryza sativa, and Marchantia polymorpha. Moreover, a mouse brain was also efficiently cleared by the iTOMEI-Brain method within 48 h, and strong fluorescent signals were detected in the cleared brain.

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参考文献

1.

2.

3.

4.

5.

6.

7.

8.

9.

10.

11.

12.

13.

14.

15.

16.

17.

18.

19.

20.

21.

Statistics and reproducibility. The statistical significance between two groups was

evaluated using a two-sided Welch’s t-test, whereas comparisons of multiple groups

Okabe, K., Sakaguchi, R., Shi, B. & Kiyonaka, S. Intracellular thermometry

with fluorescent sensors for thermal biology. Pflug. Arch. Eur. J. Physiol. 470,

717–731 (2018).

Lin, M. Z. & Schnitzer, M. J. Genetically encoded indicators of neuronal

activity. Nat. Neurosci. 19, 1142–1153 (2016).

Miyanari, Y., Ziegler-Birling, C. & Torres-Padilla, M.-E. Live visualization of

chromatin dynamics with fluorescent TALEs. Nat. Struct. Mol. Biol. 20,

1321–1324 (2013).

Hama, H. et al. ScaleS: an optical clearing palette for biological imaging. Nat.

Neurosci. 18, 1518–1529 (2015).

Susaki, E. A. et al. Whole-brain imaging with single-cell resolution using

chemical cocktails and computational analysis. Cell 157, 726–739 (2014).

Yang, B. et al. Single-cell phenotyping within transparent intact tissue through

whole-body clearing. Cell 158, 945–958 (2014).

Ke, M.-T. et al. Super-resolution mapping of neuronal circuitry with an indexoptimized clearing agent. Cell Rep. 14, 2718–2732 (2016).

Tainaka, K. et al. Chemical landscape for tissue clearing based on hydrophilic

reagents. Cell Rep. 24, 2196–2210.e9 (2018).

Warner, C. A. et al. An optical clearing technique for plant tissues allowing

deep imaging and compatible with fluorescence microscopy. Plant Physiol.

166, 1684–1687 (2014).

Kurihara, D., Mizuta, Y., Sato, Y. & Higashiyama, T. ClearSee: a rapid optical

clearing reagent for whole-plant fluorescence imaging. Development 142,

4168–4179 (2015).

Kurihara, D., Mizuta, Y., Nagahara, S. & Higashiyama, T. ClearSeeAlpha:

advanced optical clearing for whole-plant imaging. Plant Cell Physiol. 62,

1302–1310 (2021).

Palmer, W. M. et al. PEA-CLARITY: 3D molecular imaging of whole plant

organs. Sci. Rep. 5, 13492 (2015).

Hasegawa, J. et al. Three-dimensional imaging of plant organs using a simple

and rapid transparency technique. Plant Cell Physiol. 57, 462–472 (2016).

Katagiri, Y. et al. The coordination of ploidy and cell size differs between cell

layers in leaves. Development 143, 1120–1125 (2016).

Sakamoto, Y. & Matsunaga, S. Deep imaging of plant roots by a rapid

transparency technique TOMEI. CYTOLOGIA 82, 221–222 (2017).

Aoyagi, Y., Kawakami, R., Osanai, H., Hibi, T. & Nemoto, T. A rapid optical

clearing protocol using 2,2′-thiodiethanol for microscopic observation of fixed

mouse brain. PLoS ONE 10, e0116280 (2015).

Costantini, I. et al. A versatile clearing agent for multi-modal brain imaging.

Sci. Rep. 5, 9808 (2015).

Nurani, A. M. et al. Deep imaging analysis in VISUAL reveals the role of

YABBY genes in vascular stem cell fate determination. Plant Cell Physiol. 61,

255–264 (2020).

Musielak, T. J., Slane, D., Liebig, C. & Bayer, M. A versatile optical clearing

protocol for deep tissue imaging of fluorescent proteins in Arabidopsis

thaliana. PLoS ONE 11, e0161107 (2016).

Guo, W. et al. Chemical reactivation of resin-embedded pHuji adds red for

simultaneous two-color imaging with EGFP. Biomed. Opt. Express 8, 3281 (2017).

Xiong, H. et al. Chemical reactivation of quenched fluorescent protein

molecules enables resin-embedded fluorescence microimaging. Nat. Commun.

5, 3992 (2014).

COMMUNICATIONS BIOLOGY | (2022)5:12 | https://doi.org/10.1038/s42003-021-02955-9 | www.nature.com/commsbio

11

ARTICLE

COMMUNICATIONS BIOLOGY | https://doi.org/10.1038/s42003-021-02955-9

22. Schnell, U., Dijk, F., Sjollema, K. A. & Giepmans, B. N. G.

Immunolabeling artifacts and the need for live-cell imaging. Nat. Methods

9, 152–158 (2012).

23. DesMarais, V., Eddy, R. J., Sharma, V. P., Stone, O. & Condeelis, J. S.

Optimizing leading edge F-actin labeling using multiple actin probes, fixation

methods and imaging modalities. BioTechniques 66, 113–119 (2019).

24. Moldovan, G.-L., Pfander, B. & Jentsch, S. PCNA, the maestro of the

replication fork. Cell 129, 665–679 (2007).

25. Yokoyama, R., Hirakawa, T., Hayashi, S., Sakamoto, T. & Matsunaga, S.

Dynamics of plant DNA replication based on PCNA visualization. Sci. Rep. 6,

29657 (2016).

26. Tamaki, S. et al. FT-like proteins induce transposon silencing in the shoot

apex during floral induction in rice. Proc. Natl Acad. Sci. USA 112, E901–E910

(2015).

27. Shimamura, M. Marchantia polymorpha: taxonomy, phylogeny and

morphology of a model system. Plant Cell Physiol. 57, 230–256 (2016).

28. Kim, C. M., Han, C. & Dolan, L. RSL class I genes positively regulate root hair

development in Oryza sativa. N. Phytol. 213, 314–323 (2017).

29. Menand, B. et al. An ancient mechanism controls the development

of cells with a rooting function in land plants. Science 316, 1477–1480

(2007).

30. Proust, H. et al. RSL class I genes controlled the development of epidermal

structures in the common ancestor of land plants. Curr. Biol. 26, 93–99

(2016).

31. Jang, G., Yi, K., Pires, N. D., Menand, B. & Dolan, L. RSL genes are sufficient

for rhizoid system development in early diverging land plants. Development

138, 2273–2281 (2011).

32. Wu, F. et al. The ABCs of flower development: mutational analysis of AP1 /

FUL -like genes in rice provides evidence for a homeotic (A)-function in

grasses. Plant J. 89, 310–324 (2017).

33. Ikeda, K., Sunohara, H. & Nagato, Y. Developmental course of inflorescence

and spikelet in rice. Breed. Sci. 54, 147–156 (2004).

34. Taoka, K. et al. 14-3-3 proteins act as intracellular receptors for rice Hd3a

florigen. Nature 476, 332–335 (2011).

35. Hirakawa, T. et al. LSD1-LIKE1-mediated H3K4me2 demethylation is

required for homologous recombination repair. Plant Physiol. 181, 499–509

(2019).

36. Ishihara, H. et al. Primed histone demethylation regulates shoot regenerative

competency. Nat. Commun. 10, 1786 (2019).

37. Higo, A. et al. DNA methylation is reconfigured at the onset of reproduction

in rice shoot apical meristem. Nat. Commun. 11, 4079 (2020).

38. Lucob‐Agustin, N. et al. WEG1, which encodes a cell wall hydroxyproline‐rich

glycoprotein, is essential for parental root elongation controlling lateral root

formation in rice. Physiol. Plant. 169, 214–227 (2020).

39. Ishizaki, K. et al. Development of gateway binary vector series with four

different selection markers for the liverwort Marchantia polymorpha. PLoS

ONE 10, e0138876 (2015).

40. Adachi, S. et al. Programmed induction of endoreduplication by DNA doublestrand breaks in Arabidopsis. Proc. Natl Acad. Sci. USA 108, 10004–10009

(2011).

41. Ishizaki, K., Chiyoda, S., Yamato, K. T. & Kohchi, T. Agrobacteriummediated transformation of the haploid liverwort Marchantia polymorpha

L. an emerging model for plant biology. Plant Cell Physiol. 49, 1084–1091

(2008).

42. Paxinos, G., & Franklin, K. B. J. The Mouse Brain in Stereotaxic Coordinates

2nd edn (Academic Press, 2001).

12

Acknowledgements

We thank Dr. Kimitsune Ishizaki for providing the histone H2B-tdTomato entry vector

and Dr. Yuichiro Watanabe, Dr. Takahiro Hamada, Mr. Kazutaka Futagami, and Ms.

Sakiko Ishida for technical support with transformation of M. polymorpha. This research

was supported by MXT/JSPS KAKENHI 18K14743 and 21K06247 to Y. Sakamoto,

20H04884 to R.N., and 19H03259, 20H03297, and 20H05911 to S.M. Y. Sakamoto is also

supported by the Osaka University Program for the Support of Networking among

Present and Future Researchers, and S.M. is also supported by the Mitsubishi Foundation. We thank Edanz (https://jp.edanz.com/ac) for editing a draft of this manuscript.

Author contributions

Y. Sakamoto and S.M. designed the research; Y. Sakamoto, A.I., Y. Sakai, M.S., R.N., K.A.,

and Y. Sano performed the research; Y. Sakamoto and A.I. analyzed the data; Y. Sakamoto, Y. Sano, T.F., H.T., T.K., and S.M. wrote the paper.

Competing interests

The authors declare the following competing interests: the patent application in Japan.

Patent applicant: Tokyo University of Science, Inventors: Yuki Sakamoto and Sachihiro

Matsunaga, Application No.: 2020-026975, Status: pending. Yuki Sakamoto and Sachihiro Matsunaga are unpaid advisory positions of plant tissue-clearing reagents in Tokyo

Chemical Industry Cooperation (Tokyo, Japan). The remaining authors declare no

competing interests.

Additional information

Supplementary information The online version contains supplementary material

available at https://doi.org/10.1038/s42003-021-02955-9.

Correspondence and requests for materials should be addressed to Sachihiro Matsunaga.

Peer review information Communications Biology thanks Sunghoe Chang, Irene

Costantini, and Noriko Inada for their contribution to the peer review of this work.

Primary Handling Editors: Chao Zhou and Caitlin Karniski. Peer reviewer reports are

available.

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