Development of live cell imaging experimental system in babesia parasite

• Project/Area Number: 24658242
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Basic veterinary science/Basic zootechnical science
• Research Institution: Obihiro University of Agriculture and Veterinary Medicine
• Principal Investigator: KAWAZU Shin-ichiro 帯広畜産大学, 原虫病研究センター, 教授 (60312295)
• Co-Investigator(Renkei-kenkyūsha): TSUJI Naotoshi 動物衛生研究所, 細菌・寄生虫研究領域, 主任研究員 (70355171)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2012: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 獣医学 / 感染症 / バベシア / ライブイメージング / 遺伝子改変原虫

Research Abstract

The system for stable expression of green fluorescent protein (GFP) in babesia parasite was developed and the GFP-expressing parasite was observed through time-lapse video microscopy. (1) Stable expression of GFP in B. bovis using the WR99210/hDHFR selection system was achieved. The expression cassette was applied for the development of gene targeting (knockout) technique and the knockout parasite was established for the first time. (2) The gliding motility of B. bovis merozoites was revealed for the first time by in vitro time-lapse video microscopy of the GFP-expressing parasite. In vivo infection model of the parasite with SCID mice, with which a part of the circulating erythrocyte had been replaced with bovine cells, was also developed to investigate the gliding motility of the parasite in detail through time-lapse video microscopy.

Research Products

• [Journal Article] Cloning and Characterization of a 2-Cys Peroxiredoxin from <i>Babesia gibsoni</i> (2014)
  • Author(s): Masatani T, Asada M, Ichikawa-Seki M, Usui M, Terkawi MA, Hayashi K, Kawazu S, Xuan X.
  • Journal Title: Journal of Veterinary Medical Science Volume: 76 Issue: 1 Pages: 139-143
  • DOI: 10.1292/jvms.13-0274
  • NAID: 130003362301
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed
• [Journal Article] Gliding Motility of Babesia bovis Merozoites Visualized by Time-Lapse Video Microscopy (2012)
  • Author(s): Asada, M., Goto, Y., Yahata, K., Yokoyama, N., Kawai, S., Inoue, N., Kaneko, O., and Kawazu, S.-I
  • Journal Title: PLoS ONE Volume: 7 Issue: 4 Pages: e35227-e35227
  • DOI: 10.1371/journal.pone.0035227
  • Peer Reviewed
• [Presentation] バベシア原虫におけるダブルトランスフェクション法の確立 (2014)
  • Author(s): 麻田正仁, 他
  • Organizer: 第83回日本寄生虫学会
  • Place of Presentation: 愛媛大学城北キャンパス
  • Year and Date: 2014-03-27
• [Presentation] Study on gliding motility of Babesia bovis merozo ites using bioimaging analysis (2014)
  • Author(s): Asada, M., and Kawazu, S
  • Organizer: 6^<th> ASEA N Congress of Tropical Medicine and Parasitology (第6回ASEAN熱帯医学寄生虫学会議)
  • Place of Presentation: Intercontinental Hotel Kuala Lumpur(クアラルンプール、マレーシア)
  • Year and Date: 2014-03-07
• [Presentation] バベシア原虫におけるダブルトランスフェクション法の確立 (2014)
  • Author(s): 麻田正仁、Carlos Suarez、横山直明、五十嵐郁男、矢幡一英、金子修、河津信一郎
  • Organizer: 第83回日本寄生虫学会
  • Place of Presentation: 愛媛大学 城北キャンパス
• [Presentation] Study on gliding motility of Babesia bovis merozoites using bioimaging analysis. (2014)
  • Author(s): Asada, M., and Kawazu, S.
  • Organizer: 6th ASEAN Congress of Tropical Medicine and Parasitology 第6回ASEAN熱帯医学寄生虫学会議
  • Place of Presentation: Intercontinental Hotel Kuala Lumpur（クアラルンプール、マレーシア）
  • Invited
• [Presentation] Babesia bovisチオレドキシンペルオキシターゼBbTPx-1ノックアウト原虫は活性窒素種負荷に対する感受性が上昇する (2013)
  • Author(s): 麻田正仁, 他
  • Organizer: 第156回日本獣医学会学術集会
  • Place of Presentation: 岐阜大学
  • Year and Date: 2013-09-21
• [Presentation] Babesia bovisチオレドキシンペルオキシターゼBbTPx-1ノックアウト原虫の活性窒素種負荷に対する感受性上昇 (2013)
  • Author(s): 麻田正仁、Carlos Suarez、薄井美帆、後藤康之、横山直明、井上 昇、矢幡一英、金子修、河津信一郎
  • Organizer: 第11回分子寄生虫学マラリア研究フォーラム
  • Place of Presentation: 長崎大学、熱帯医学研究所
• [Presentation] バイオイメージングによるBabesia bovis メロゾイト滑走運動の研究 (2013)
  • Author(s): 麻田正仁 他
  • Organizer: 第82回日本寄生虫学会大会
  • Place of Presentation: 東京医科歯科大学 湯島キャンパス
  • Invited
• [Presentation] Gliding motility of Babesia bovis merozoites visualized by time-lapse video microscopy (2012)
  • Author(s): Asada, M., et al.
  • Organizer: American Society of Tropical Medicine and Hygie ne 61^<th> Annual Meeting (第61回米国熱帯医学会年会)
  • Place of Presentation: Atlanta Marriot Marquis(アトランタ、米国)
  • Year and Date: 2012-10-12
• [Presentation] Babesia bovis メロゾイト滑走運動のバイオイメージング解析 (2012)
  • Author(s): 麻田正仁, 他
  • Organizer: 第10回分子寄生虫学マラリア研究フォーラム
  • Place of Presentation: 群馬大学医学部
  • Year and Date: 2012-10-12
• [Presentation] Gliding motility of Babesia bovis merozoites visualized by time-lapse video microscopy. (2012)
  • Author(s): Asada, M. et al.
  • Organizer: American Society of Tropical Medicine and Hygiene 61th Annual Meeting 第61回米国熱帯医学会年会
  • Place of Presentation: Atlanta Marriot Marquis（アトランタ、米国）
• [Presentation] Babesia bovis メロゾイト滑走運動のバイオイメージング解析 (2012)
  • Author(s): 麻田正仁 他
  • Organizer: 第10回分子寄生虫学マラリア研究フォーラム
  • Place of Presentation: 群馬大学 医学部
• [Presentation] H2O2蛍光プローブを用いたバベシア原虫細胞内酸化ストレスの可視化 (2012)
  • Author(s): 麻田正仁 他
  • Organizer: 第154回日本獣医学会学術集会
  • Place of Presentation: 岩手大学
• [Remarks]
  • URL: http://www.obihiro.ac.jp/~tryp/index.html
• [Remarks] Kawazu Laboratory, NRCPD
  • URL: http://www.obihiro.ac.jp/~tryp/index.html
• [Remarks] 河津研究室ホームページ
  • URL: http://www.obihiro.ac.jp/~tryp/index.html