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A study on MTs in the cell cortex by HP freezing

Research Project

Project/Area Number 09044225
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section一般
Research Field 生物形態・構造
Research InstitutionHiroshima University

Principal Investigator

MINEYUKI Yoshinobu  HIROSHIMA UNIVERSITY, FACULTY OF SCIENCE, ASSOCIATE PROFESSOR, 理学部, 助教授 (30219703)

Co-Investigator(Kenkyū-buntansha) KARAHARA Ichirou  TOYAMA UNIVERSITY, FACULTY OF SCIENCE, RESEARCH ASSOCIATE, 理学部, 助手 (60283058)
MURATA Takashi  UNIVERSITY OF TOKYO, GRADUATE SCHOOL OF ARTS AND SCIENCE, ASSISTANT PROFESSOR, 大学院・総合文化研究科, 講師 (00242024)
GIDDINGS Jr  コロラド大学, 分子細胞発生生物学科電子顕微鏡ユニット, 主任
STAEHELIY L.  コロラド大学, 分子細胞発生生物学科, 教授
GIDDINGS.JR トーマス  コロラド大学, 分子細胞発生生物学科・電子顕微鏡ユニット, 主任
STAEHELIN L.  コロラド大学, 分子細胞発生生物学科, 教授
Project Period (FY) 1997 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥8,600,000 (Direct Cost: ¥8,600,000)
Fiscal Year 1999: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1998: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1997: ¥2,900,000 (Direct Cost: ¥2,900,000)
KeywordsHigh pressure freezing / Electron microscope / Microtubules / preprophase band / Cytokinesis / Coated vesicle / Root meristems / Tomography / 加圧凍結装置 / 休息凍結法 / 急速凍結法
Research Abstract

Interphase cortical microtubules (MTs) and the preprophase band (PPB) are two cortical arrays of MTs in higher plants, and are known to play ezzentrrial role for the regulation of the direction of cell elongation and for the determination of the division site, respectively. However, little is known about the mechanism that regulates these cortical MT organization and turnover. We have employed a combination of high pressure freezing and freeze-substitution or freeze-etch techniques to preserve and visualize forming MTs in onion and tobacco seedlings for ultrastructural analysis. These studies have led to the detection of several novel structural features that have not been reported previously.
(1) In vitro studies of polymerizing/depolymerizing MTs have shown that their ends display both flared and non-flared configurations. Our micrographs demonstrate that cortical MTs also exhibit flared and non-flared ends, consistent with the idea that they are turning over. The number of coiled end configurations (expected depolymerizing MT end in vivo) increased by the treatment with a MT-depolymerizing drug, oryzalin. This suggests that the the frequency of coiled ends reflects that of shrinklng MTs in living cells. (2) Drugs that affect F-actin stability have been shown to perturb PPB morphology. To learn more about how such drugs may cause these changes, we are analyzing quantitatively their effects on both MT-MT and MT-plasma membrane spacings. This type of analysis is more easily done with cryofixed than with chemically fixed samples due to the smooth, non-wavy conformation of the plasma membrane in high pressure frozen cells. (3) Previous studies have shown that PPB regions contain vesicles that stain similar to cell walls, which has led to the suggestion that they could be Involved in delivering special types of cell wall molecules to PPB regions. Our micrographs demonstrate, however, that increased numbers of coated, endocytic vesicles are formed between the PPB MTs.

Report

(4 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] 峰雪芳宣: "Preprophase bandの訳語について"Plant Morphology. 9. 65-68 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 峰雪芳宣: "加圧凍結法による高等植物実生分裂組織細胞の観察"Plant Morphology. 10. 30-39 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Y.Mineyuki: "The Preprophase band of microtubules:Its function as a cytokinetic apparatus in higher plants"International Review for Cytology. 187. 1-49 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] A.Nogami: "Loosening of a preprophase band of microtubules in onion(Allium cepa L.) root tip"Cell Structure and Function. 24. 419-424 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Mineyuki, Y.: "About the term Preprophase band in Japanese"Plant Morphology. 9. 65-68 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Mineyuki. Y.: "Observation of meristematic cells in seedlings of higher plants using a high pressure freezing methods."Plant Morphology. 10. 30-39 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Mineyuki. Y.: "The preprophase band of microtubules : Its function as a cytokinetic apparatus in higher plants."International Review of Cytology. 187. 1-49 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nogami, A.: "Loosening of a preprophase band of microtubules in onion root tip cells by kinase inhibitors."Cell Structure and Function. 24. 419-424 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Y.Mineyuki: "The preprophase band of microtubules:Its function as a cytokinetic apparatus in higher plants."International Review for Cytology. 187. 1-49 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] A.Nogami: "Loosening of a preprophase band of microtubules in onion(Allium cepa L.)root tip cells by kinase inhibitors."Cell Structure and Function. 24. 419-424 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 峰雪芳宣、村田隆、T.H.Giddings, L.A.Steahelin: "加圧凍結法による高等植物実生分裂組織細胞の観察" Plant Morphology. 10・1. 30-39 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Y.Mineyuki: "The preprophase band of microtubules : Its function as a cytokinetic apparatus in higher plants" International Revies for Cytology. 187. 1-49 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] 峰雪 芳宣: "Preprophase bandの訳語について" Plant Morphology. 9・1. 65-68 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Y.Mineyuki: "The preprophase band of microtubules:Its function as a cytokinetic apparatus in higher plants." International Review for Cytology. (印刷中). (1998)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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