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Journal of Plant Research

, Volume 126, Issue 1, pp 113–119 | Cite as

Microtubule stability affects the unique motility of F-actin in Marchantia polymorpha

  • Atsuko Era
  • Natsumaro Kutsuna
  • Takumi Higaki
  • Seiichiro Hasezawa
  • Akihiko Nakano
  • Takashi UedaEmail author
Regular Paper

Abstract

Actin microfilaments play crucial roles in diverse plant functions. Some specific cellular processes require interaction between F-actin and microtubules, and it is believed that there are direct or indirect connections between F-actin and microtubules. We previously reported that actin microfilaments exhibit unique dynamic motility in cells of the liverwort, Marchantia polymorpha; the relevance of this activity to microtubules has not been explored. To examine whether the dynamics of F-actin in M. polymorpha were somehow regulated by microtubules, we investigated the effects of stabilization or destabilization of microtubules on dynamics of actin bundles, which were visualized by Lifeact-Venus. To our surprise, both stabilization and destabilization of microtubules exerted similar effects on F-actin motility; apparent sliding movement of F-actin in M. polymorpha cells was accelerated by both oryzalin and paclitaxel, with the effect of paclitaxel more evident than that of oryzalin. Immunofluorescence staining revealed that some F-actin bundles were arrayed along with microtubules in M. polymorpha thallus cells. These results suggest that microtubules play regulatory roles in the unique F-actin dynamics in M. polymorpha.

Keywords

Actin Image analysis Lifeact Marchantia polymorpha Microtubule 

Notes

Acknowledgments

We would like to thank T. Kohchi and K. Ishizaki (Kyoto University), and K.T. Yamato (Kinki University) for their generous support for M. polymorpha studies. This work was supported by Grants-in-Aid for Scientific Research and the Targeted Proteins Research Program (TPRP) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, and Grant-in-Aid for JSPS Fellows (A.E.). This research was also supported by JST, PRESTO.

Supplementary material

10265_2012_496_MOESM1_ESM.tif (1.5 mb)
Supplementary material 1 (TIFF 1538 kb) Workflow of image processing for calculation of the movement index. (a) An original time-sequential image O at time point t. (b) A subtracted image D obtained from the original image (a). (c) A shift-diffuse filtered image of the subtracted image (b). (d) A thresholded image, M obtained from the shift-diffuse filtered image (c). (e) An AF-segmented image, T obtained from the original images (a). (f) An image, U for calculation of the movement index of AFs obtained from the skeletonized image (e) and the thresholded image (d). (g) An image, U at time point t + 1. (h) A merged image of (f) (red) and (g) (green). Note that apparent sliding movement was much more obvious than lateral movement. See Materials and methods for algorithms for determination of the measurements
10265_2012_496_MOESM2_ESM.tif (1.7 mb)
Supplementary material 2 (TIFF 1699 kb) Immunofluorescent staining of MTs in the oryzalin treated M. polymorpha cell. Maximum intensity projection of z-stack images. MTs were completely disrupted. Scale bar = 20 μm
10265_2012_496_MOESM3_ESM.mov (475 kb)
Supplementary material 3 (MOV 475 kb) Dynamics of actin bundles in a M. polymorpha cell treated with DMSO (control). Pictures were taken every second and are shown at 10-fold increased speed
10265_2012_496_MOESM4_ESM.mov (222 kb)
Supplementary material 4 (MOV 222 kb) Dynamics of actin bundles in a M. polymorpha cell treated with BDM. Pictures were taken every second and are shown at 10-fold increased speed
10265_2012_496_MOESM5_ESM.mov (195 kb)
Supplementary material 5 (MOV 195 kb) Dynamics of actin bundles in a M. polymorpha cell treated with oryzalin treated. Pictures were taken every second and are shown at 10-fold increased speed
10265_2012_496_MOESM6_ESM.mov (217 kb)
Supplementary material 6 (MOV 216 kb) Dynamics of actin bundles in a M. polymorpha cell treated with paclitaxel. Pictures were taken every second and are shown at 10-fold increased speed

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Copyright information

© The Botanical Society of Japan and Springer 2012

Authors and Affiliations

  • Atsuko Era
    • 1
  • Natsumaro Kutsuna
    • 4
  • Takumi Higaki
    • 4
    • 5
  • Seiichiro Hasezawa
    • 4
    • 5
  • Akihiko Nakano
    • 1
    • 3
  • Takashi Ueda
    • 1
    • 2
    Email author
  1. 1.Department of Biological Sciences, Graduate School of ScienceThe University of TokyoTokyoJapan
  2. 2.Japan Science and Technology Agency (JST), PRESTOKawaguchiJapan
  3. 3.Molecular Membrane Biology LaboratoryRIKEN Advanced Science InstituteWakoJapan
  4. 4.Department of Integrated Biosciences, Graduate School of Frontier SciencesThe University of TokyoKashiwaJapan
  5. 5.Advanced Measurement and AnalysisJapan Science and Technology Agency (JST)TokyoJapan

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