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Multi-dimensional bioimage analyses with machine learning to reveal the dynamics of membrane vesicles and microtubules in stomatal guard cells

Research Project

Project/Area Number 20H03289
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 44040:Morphology and anatomical structure-related
Research InstitutionKumamoto University

Principal Investigator

Higaki Takumi  熊本大学, 大学院先端科学研究部(理), 教授 (90578486)

Project Period (FY) 2020-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥16,120,000 (Direct Cost: ¥12,400,000、Indirect Cost: ¥3,720,000)
Fiscal Year 2022: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2021: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2020: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Keywords気孔開閉運動 / 微小管 / 膜小胞 / イメージング / 画像解析
Outline of Research at the Start

本研究では、多次元ライブセルイメージングと機械学習(いわゆる人工知能(AI))を活用した画像解析技術に基づいて、植物の気孔開閉運動を司るH+-ATPaseの細胞膜への局在化機構の全貌解明を目指す。気孔開口を駆動するH+-ATPase AHA1の細胞膜輸送に必須の因子であるPATROL1およびその相互作用因子群に注目し、気孔開閉運動におけるPATROL1関連因子群の三次元分布および共局在性を描出する。取得画像群から膜小胞と微小管を高精度に検出・評価する機械学習技術を活用した次世代型の定量的細胞生物学研究を展開し、気孔応答を制御するPATROL1依存的なAHA1の細胞膜輸送機構を明らかにする。

Outline of Final Research Achievements

In this study, we aimed to elucidate the mechanism of H+-ATPase plasma membrane delivery through next-generation quantitative cell biology research by leveraging AI technology to dramatically reduce the human cost associated with image analysis of multidimensional live cell imaging, a bottleneck until now. Specifically, we (1) explored the application of image transformations using deep learning for segmentation, an indispensable task for quantitatively evaluating the cytoskeletal structure; (2) investigated the improvement of phototoxicity and time resolution through the enhancement of microscopic image quality by deep learning; (3) automated the classification and analysis of cytoskeletal patterns through machine learning; and (4) using these technologies, we conducted dynamic analysis related to drug resistance of membrane vesicles and superficial microtubules that control stomatal movement.

Academic Significance and Societal Importance of the Research Achievements

本研究では、AI技術を利用して植物細胞生物学研究を推進する方法を実践的に検討した。具体的には、細胞骨格構造の定量評価を改善するセグメンテーション手法、顕微鏡画像の画質改善を目指した深層学習モデルの開発、細胞骨格パターンの自動分類と分析など、研究者の分析作業を省力化するだけでなく、従来よりもより健全な状況の細胞をより詳しく調べることのできる技術の開発に取り組んだ。さらに、これらの手法を用いて植物の孔辺細胞に焦点を当て、気孔の環境応答性を担うH+-ATPase細胞膜輸送機構の解明を目指して実践的な研究を行った。このAIを活用した手法は、生物学の幅広い分野での利用が期待される。

Report

(4 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Annual Research Report
  • 2020 Annual Research Report
  • Research Products

    (37 results)

All 2023 2022 2021 2020 Other

All Int'l Joint Research (1 results) Journal Article (25 results) (of which Int'l Joint Research: 9 results,  Peer Reviewed: 25 results,  Open Access: 23 results) Presentation (7 results) (of which Int'l Joint Research: 2 results,  Invited: 5 results) Remarks (4 results)

  • [Int'l Joint Research] UC Davis/Michigan State University(米国)

    • Related Report
      2020 Annual Research Report
  • [Journal Article] Ab-GALFA, A bioassay for insect gall formation using the model plant Arabidopsis thaliana2023

    • Author(s)
      Tomoko Hirano , Ayaka Okamoto , Yoshihisa Oda , Tomoaki Sakamoto , Seiji Takeda , Takakazu Matsuura , Yoko Ikeda , Takumi Higaki , Seisuke Kimura , Masa H Sato
    • Journal Title

      Scientific Reports

      Volume: 13 Issue: 1 Pages: 2554-2554

    • DOI

      10.1038/s41598-023-29302-8

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Migration of prospindle before the first asymmetric division in germinating spore of <i>Marchantia polymorpha</i>2022

    • Author(s)
      Sakai Yuuki、Higaki Takumi、Ishizaki Kimitsune、Nishihama Ryuichi、Kohchi Takayuki、Hasezawa Seiichiro
    • Journal Title

      Plant Biotechnology

      Volume: 39 Issue: 1 Pages: 5-12

    • DOI

      10.5511/plantbiotechnology.21.1217b

    • ISSN
      1342-4580, 1347-6114
    • Year and Date
      2022-03-25
    • Related Report
      2022 Annual Research Report 2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Tracking metabolites at single-cell resolution reveals metabolic dynamics during plant mitosis.2022

    • Author(s)
      Okubo-Kurihara E, Ali A, Hiramoto M, Kurihara Y, Abouleila Y, Abdelazem EM, Kawai T, Makita Y, Kawashima M, Esaki T, Shimada H, Mori T, Hirai MY, Higaki T, Hasezawa S, Shimizu Y, Masujima T, Matsui M
    • Journal Title

      Plant Physiol

      Volume: 189 Issue: 2 Pages: 459-464

    • DOI

      10.1093/plphys/kiac093

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Two Kinesin-14A motors oligomerize to drive poleward microtubule convergence for acentrosomal spindle morphogenesis in Arabidopsis thaliana.2022

    • Author(s)
      Hotta T, Lee YR, Higaki T, Hashimoto T, Liu B
    • Journal Title

      Front Cell Dev Biol.

      Volume: 10 Pages: 949345-949345

    • DOI

      10.3389/fcell.2022.949345

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Two-step regulation of centromere distribution by condensin II and the nuclear envelope proteins2022

    • Author(s)
      Sakamoto Takuya、Sakamoto Yuki、Grob Stefan、Slane Daniel、Yamashita Tomoe、Ito Nanami、Oko Yuka、Sugiyama Tomoya、Higaki Takumi、Hasezawa Seiichiro、Tanaka Maho、Matsui Akihiro、Seki Motoaki、Suzuki Takamasa、Grossniklaus Ueli、Matsunaga Sachihiro
    • Journal Title

      Nature Plants

      Volume: 8 Issue: 8 Pages: 940-953

    • DOI

      10.1038/s41477-022-01200-3

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Bone marrow imaging reveals the migration dynamics of neonatal hematopoietic stem cells2022

    • Author(s)
      Takihara Yuji、Higaki Takumi、Yokomizo Tomomasa、Umemoto Terumasa、Ariyoshi Kazunori、Hashimoto Michihiro、Sezaki Maiko、Takizawa Hitoshi、Inoue Toshihiro、Suda Toshio、Mizuno Hidenobu
    • Journal Title

      Communications Biology

      Volume: 5 Issue: 1 Pages: 776-776

    • DOI

      10.1038/s42003-022-03733-x

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] EML2-S constitutes a new class of proteins that recognizes and regulates the dynamics of tyrosinated microtubules.2022

    • Author(s)
      Hotta T, McAlear TS, Yue Y, Higaki T, Haynes SE, Nesvizhskii AI, Sept D, Verhey KJ, Bechstedt S, Ohi R
    • Journal Title

      Curr Biol

      Volume: 32 Issue: 18 Pages: 3898-3910

    • DOI

      10.1016/j.cub.2022.07.027

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Machine learning and feature analysis of the cortical microtubule organization of Arabidopsis cotyledon pavement cells.2022

    • Author(s)
      Yoshida D, Akita K, Higaki T
    • Journal Title

      Protoplasma

      Volume: - Issue: 3 Pages: 987-998

    • DOI

      10.1007/s00709-022-01813-7

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Disease-associated H58Y mutation affects the nuclear dynamics of human DNA topoisomerase IIβ.2022

    • Author(s)
      Morotomi-Yano K, Hiromoto Y, Higaki T, Yano KI
    • Journal Title

      Sci Rep

      Volume: 12 Issue: 1 Pages: 20627-20627

    • DOI

      10.1038/s41598-022-24883-2

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Calcium signaling contributes to xylem vessel cell differentiation via post-transcriptional regulation of VND7 downstream events2021

    • Author(s)
      Kamon E, Noda C, Higaki T, Demura T, Ohtani M
    • Journal Title

      Plant Biotechnology

      Volume: 38 Issue: 3 Pages: 331-337

    • DOI

      10.5511/plantbiotechnology.21.0519a

    • NAID

      130008091606

    • ISSN
      1342-4580, 1347-6114
    • Year and Date
      2021-09-25
    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] The boundary-expressed <i>EPIDERMAL PATTERNING FACTOR-LIKE2</i> gene encoding a signaling peptide promotes cotyledon growth during <i>Arabidopsis thaliana</i> embryogenesis2021

    • Author(s)
      Fujihara Rina、Uchida Naoyuki、Tameshige Toshiaki、Kawamoto Nozomi、Hotokezaka Yugo、Higaki Takumi、Simon Rudiger、Torii Keiko U、Tasaka Masao、Aida Mitsuhiro
    • Journal Title

      Plant Biotechnology

      Volume: 38 Issue: 3 Pages: 317-322

    • DOI

      10.5511/plantbiotechnology.21.0508a

    • NAID

      130008091603

    • ISSN
      1342-4580, 1347-6114
    • Year and Date
      2021-09-25
    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Involvement of the Membrane Trafficking Factor PATROL1 in the Salinity Stress Tolerance of <i>Arabidopsis thaliana</i>2021

    • Author(s)
      Sato F, Iba K, Higaki T
    • Journal Title

      CYTOLOGIA

      Volume: 86 Issue: 2 Pages: 119-126

    • DOI

      10.1508/cytologia.86.119

    • NAID

      130008056667

    • ISSN
      0011-4545, 1348-7019
    • Year and Date
      2021-06-25
    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] PUCHI Regulates Giant Cell Morphology During Root-Knot Nematode Infection in Arabidopsis thaliana2021

    • Author(s)
      Suzuki Reira、Yamada Mizuki、Higaki Takumi、Aida Mitsuhiro、Kubo Minoru、Tsai Allen Yi-Lun、Sawa Shinichiro
    • Journal Title

      Frontiers in Plant Science

      Volume: 12 Pages: 755610-755610

    • DOI

      10.3389/fpls.2021.755610

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Metal-nano-ink coating for monitoring and quantification of cotyledon epidermal cell morphogenesis.2021

    • Author(s)
      Kikukawa K, Yoshimura K, Watanabe A, Higaki T
    • Journal Title

      Front Plant Sci

      Volume: 12 Pages: 745980-745980

    • DOI

      10.3389/fpls.2021.745980

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] The phosphorylation status of NPH3 affects photosensory adaptation during the phototropic response.2021

    • Author(s)
      Kimura T, Haga K, Nomura Y, Higaki T, Nakagami H, Sakai T
    • Journal Title

      Plant Physiol

      Volume: 187 Issue: 2 Pages: 981-995

    • DOI

      10.1093/plphys/kiab281

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Dynamic Rearrangement and Directional Migration of Tubular Vacuoles are Required for the Asymmetric Division of the <i>Arabidopsis</i> Zygote2021

    • Author(s)
      Matsumoto Hikari、Kimata Yusuke、Higaki Takumi、Higashiyama Tetsuya、Ueda Minako
    • Journal Title

      Plant and Cell Physiology

      Volume: 62 Issue: 8 Pages: 1280-1289

    • DOI

      10.1093/pcp/pcab075

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] A three-dimensional scanning system for digital archiving and quantitative evaluation of Arabidopsis plant architectures.2021

    • Author(s)
      Kunita I, Morita MT, Toda M, Higaki T
    • Journal Title

      Plant Cell Physiol

      Volume: 62 Issue: 12 Pages: 1975-1982

    • DOI

      10.1093/pcp/pcab068

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Disruption of actin filaments delays accumulation of cell plate membranes after chromosome separation.2021

    • Author(s)
      Maeda K, Higaki T
    • Journal Title

      Plant Sig Behav

      Volume: 16 Issue: 4 Pages: 1873586-1873586

    • DOI

      10.1080/15592324.2021.1873586

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Discovery, characterization and functional improvement of kumamonamide as a novel plant growth inhibitor that disturbs plant microtubules.2021

    • Author(s)
      Ishida T, Yoshimura H, Takekawa M, Higaki T, Ideue T, Hatano M, Igarashi M, Tani T, Sawa S, Ishikawa H.
    • Journal Title

      Sci Rep

      Volume: 23 Issue: 1 Pages: 6077-6077

    • DOI

      10.1038/s41598-021-85501-1

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Four-dimensional imaging with virtual reality to quantitatively explore jigsaw puzzle-like morphogenesis of <i>Arabidopsis</i> cotyledon pavement cells2020

    • Author(s)
      Higaki T, Mizuno H
    • Journal Title

      Plant Biotechnology

      Volume: 37 Issue: 4 Pages: 429-435

    • DOI

      10.5511/plantbiotechnology.20.0605a

    • NAID

      130007960223

    • ISSN
      1342-4580, 1347-6114
    • Year and Date
      2020-12-25
    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Elevated CO2 promotes satellite stomata production in young cotyledons of Arabidopsis thaliana.2020

    • Author(s)
      Higaki T, Akita K, Hasezawa S
    • Journal Title

      Genes Cells

      Volume: 25 Issue: 7 Pages: 475-482

    • DOI

      10.1111/gtc.12773

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Expresion of a fungal exo-β-1,3-galactanase in Arabidopsis reveals a role of type II arabinogalactans in the regulation of cell shape.2020

    • Author(s)
      Yoshimi Y, Hara K, Yoshimura M, Tanaka N, Higaki T, Tsumuraya Y, Kotake T
    • Journal Title

      J Exp Bot

      Volume: 71 Issue: 18 Pages: 5414-5424

    • DOI

      10.1093/jxb/eraa236

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Mitochondrial dynamics and segregation during the asymmetric division of Arabidopsis zygotes2020

    • Author(s)
      Kimata Yusuke、Higaki Takumi、Kurihara Daisuke、Ando Naoe、Matsumoto Hikari、Higashiyama Tetsuya、Ueda Minako
    • Journal Title

      Quantitative Plant Biology

      Volume: 1

    • DOI

      10.1017/qpb.2020.4

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Arabidopsis calcium-dependent protein kinase 3 regulates actin cytoskeleton organization and immunity.2020

    • Author(s)
      Lu YJ, Li P, Shimono M, Corrion A, Higaki T, He SY, Day B
    • Journal Title

      Nat Commun

      Volume: 11 Issue: 1 Pages: 6234-6234

    • DOI

      10.1038/s41467-020-20007-4

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Coefficient of variation as an image-intensity metric for cytoskeleton bundling.2020

    • Author(s)
      Higaki T, Akita K, Katoh K
    • Journal Title

      Sci Rep

      Volume: 10 Issue: 1 Pages: 22187-22187

    • DOI

      10.1038/s41598-020-79136-x

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] 画像解析に基づく細胞の定量評価と認識2022

    • Author(s)
      檜垣 匠
    • Organizer
      日本農薬学会第48回大会
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] Four-dimensional digital archiving and quantitative evaluation of Arabidopsis architectures.2022

    • Author(s)
      Takumi Higaki
    • Organizer
      International symposium on “Plant-Structure-Optimization”
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 植物細胞生物学を起点とした画像の定量評価と機械学習2021

    • Author(s)
      檜垣 匠
    • Organizer
      第30回日本バイオイメージング学会
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] Three-dimensional reconstruction of Arabidopsis plant architecture.2021

    • Author(s)
      Takumi Higaki
    • Organizer
      Special Satellite Event “Plant Structure Optimization” in the 7th International Conference of Plant Cell Wall Biology
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 画像解析技術の援用による植物細胞骨格研究2021

    • Author(s)
      檜垣匠
    • Organizer
      第62回日本植物生理学会年会
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] プロトンポンプAHA1の細胞膜輸送を担うPATROL1と表層微小管2020

    • Author(s)
      佐藤史弥、橋本(杉本)美海、佐藤雅彦、射場厚、馳澤盛一郎、檜垣匠
    • Organizer
      日本植物学会第84回大会
    • Related Report
      2020 Annual Research Report
  • [Presentation] プロトンポンプAHA1の細胞膜輸送を担うPATROL1と相互作用因子2020

    • Author(s)
      佐藤史弥、橋本(杉本)美海、佐藤雅彦、射場厚、門田慧奈、馳澤盛一郎、檜垣匠
    • Organizer
      第29回 日本バイオイメージング学会学術集会
    • Related Report
      2020 Annual Research Report
  • [Remarks] 熊本大学ニュース「細胞の骨組みの特徴に基づいたAIによる細胞の自動分類と分析」

    • URL

      https://www.kumamoto-u.ac.jp/whatsnew/sizen/20221020

    • Related Report
      2022 Annual Research Report
  • [Remarks] 熊本大学『フィロソフィアの扉』第31回「国際先端科学技術研究機構」

    • URL

      https://www.youtube.com/watch?v=4Nr4uK-WAH4

    • Related Report
      2020 Annual Research Report
  • [Remarks] 熊本大学ニュース: 細胞の骨組みの状態を高感度に測る技術を開発

    • URL

      https://www.kumamoto-u.ac.jp/whatsnew/sizen/20201221

    • Related Report
      2020 Annual Research Report
  • [Remarks] EurekAlert! Science News Releases

    • URL

      https://www.eurekalert.org/pub_releases/2021-01/ku-ahs011421.php

    • Related Report
      2020 Annual Research Report

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Published: 2020-04-28   Modified: 2024-01-30  

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