Mechanisms for generating positional information and cell movements during early embryogenesis in the hemimetabolous insect (a cricket)

2013 Fiscal Year Final Research Report

• Project/Area Number: 23687033
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: Developmental biology
• Research Institution: The University of Tokushima
• Principal Investigator: MITO TARO 徳島大学, ソシオテクノサイエンス研究部, 助教 (80322254)
• Project Period (FY): 2011-04-01 – 2014-03-31
• Keywords: 胚発生 / 昆虫 / 細胞移動 / 位置情報

Research Abstract

We studied mechanisms of early embryogenesis in a cricket, a model system of hemimetabolous insects. We performed RNAi analyses for components of signaling pathways, such as Wnt and BMP pathways, using GFP-expressing transgenic embryos for visualizing cellular dynamics. We found that Wnt signaling is involved in regulation of direction of movements of presumptive embryonic cells, and that BMP signaling is required for cell migration from the dorsal side towards the ventral side. We also showed that these cell signalings are involved in regulation of expression of pattern formation genes, such as otd. On the other hand, we developed gene knock-out and knock-in techniques using genome editing technologies in the cricket. In addition to progress of whole genome sequencing, these techniques provides us strong tools for further studies based on various strategies.

Research Products

• [Journal Article] Gene knockout by targeted mutagenesis in a hemimetabolous insect, the two-spotted cricket Gryllus bimaculatus, using TALENs (2014)
  • Author(s): Watanabe T, Noji S, Mito T
  • Journal Title: Methods Volume: (in press)
  • Peer Reviewed
• [Journal Article] コオロギにおけるZFN,TALEN,CRISPR/Cas9を用いた遺伝子改変 (2014)
  • Author(s): 渡辺崇人,三戸太郎,大内淑代,野地澄晴
  • Journal Title: 実験医学 Volume: (別冊)Apr Pages: 149-158
• [Journal Article] Targeted gene delivery in the cricket brain, using in vivo electroporation (2013)
  • Author(s): Matsumoto CS, Shidara H, Matsuda K, Nakamura T, Mito T, Matsumoto Y, Oka K, Ogawa H.
  • Journal Title: J Insect Physiol Volume: Oct;59(12) Pages: 1235-1241
  • Peer Reviewed
• [Journal Article] Extavour CG Developmental gene discovery in a hemimetabolous insect : de novo assembly and annotation of a transcriptome for the cricket Gryllus bimaculatus (2013)
  • Author(s): Zeng V, Ewen-Campen B, Horch HW, Roth S, Mito T
  • Journal Title: PLoS ONE Volume: May;8(5) Pages: e61479
  • Peer Reviewed
• [Journal Article] ZFN/TALENを用いたコオロギの遺伝子ノックアウト (2013)
  • Author(s): 渡辺崇人,三戸太郎,野地澄晴
  • Journal Title: 細胞工学 Volume: Apr.;32(5) Pages: 543-549
• [Journal Article] Analysis of RNA-Seq data reveals involvement of JAK/STAT signalling during leg regeneration in the cricket Gryllus bimaculatus (2013)
  • Author(s): Bando T, Ishimaru Y, Kida T, Hamada Y, Matsuoka Y, Nakamura T, Ohuchi H, Noji S, Mito T
  • Journal Title: Development Volume: Mar;140(5) Pages: 959-964
  • Peer Reviewed
• [Journal Article] Non-transgenic genome modifications in a hemimetabolous insect using zinc-finger and TAL effector nucleases (2012)
  • Author(s): Watanabe T, Ochiai H, Sakuma T, Horch HW, Hamaguchi N, Nakamura T, Bando T, Ohuchi H, Yamamoto T, Noji S, Mito T
  • Journal Title: Nature Communications Volume: Aug;3 Pages: 1017-1025
  • Peer Reviewed
• [Journal Article] Functional analysis of the role of eyes absent and sine oculis in the developing eye of the cricket Gryllus bimaculatus (2012)
  • Author(s): Takagi A, Kurita K, Terasawa T, Nakamura T, Bando T, Moriyama Y, Mito T, Noji S, Ohuchi H
  • Journal Title: Dev Growth Differ Volume: Feb;54 Pages: 227-240
  • Peer Reviewed
• [Journal Article] Ancestral functions of Delta/Notch signaling in the formation of body and leg segments in the cricket Gryllus bimaculatus (2011)
  • Author(s): Mito T, Shinmyo Y, Kurita K, Nakamura T, Ohuchi H, Noji S
  • Journal Title: Development Volume: Sep;138 Pages: 3823-3833
  • Peer Reviewed
• [Journal Article] Cricket body size is altered by systemic RNAi against insulin signaling components and epidermal growth factor receptor (2011)
  • Author(s): Dabour N, Bando T, Nakamura T, Miyawaki K, Mito T, Ohuchi H, Noji S
  • Journal Title: Dev Growth Differ Volume: Sep;53 Pages: 857-869
  • Peer Reviewed
• [Presentation] Genome modificati on technology in the cricket Gryllu s bimaculatus using a CRIPR/Cas sys tem (2014)
  • Author(s): Taro Mito et al
  • Organizer: 1st Asian Invertebrate Immunit y Symposium
  • Place of Presentation: Pusan Na tional University (Busan, Korea)
  • Year and Date: 2014-02-15
• [Presentation] Gryllus bimaculat us -a hemimetabolous insect model f or functional genomics (2013)
  • Author(s): Taro Mito et al
  • Organizer: iBeetle symposium "New horizons in molecular Z oology"
  • Place of Presentation: Georg Augu st University (Göttingen, Germany)
  • Year and Date: 20130325-26
• [Presentation] Genome modificati on in a hemimetabolous insect Gryll us bimaculatus (2013)
  • Author(s): Taro Mito et al
  • Organizer: Janelia Workshop o n Genomic Modification in Model and Non-Model Insects
  • Place of Presentation: HHMI Janelia Farm Research Campu s (Ashburn,USA)
  • Year and Date: 20130321-22
• [Presentation] ゲノム編集技術によるノックアウトコオロギの作製 (2013)
  • Author(s): 三戸太郎他
  • Organizer: 第36回日本分子生物学会年会(ワークショップ;分子生物・生命情報)
  • Place of Presentation: 神戸ポートアイランド(兵庫県神戸市)
  • Year and Date: 2013-12-04
• [Presentation] Exploring mechani sms of embryonic patterning in Gryl lus bimaculatus, a hemimetablous in sect model system (2012)
  • Author(s): Taro Mito et al
  • Organizer: 24th Internation al Congress of Entomology
  • Place of Presentation: Hotel Inre-Burgo Exco (Daegu, Korea)
  • Year and Date: 2012-08-21
• [Presentation] Exploring molecular mech anisms of early embryogenesis in th e cricket Gryllus bimaculatus (2011)
  • Author(s): Taro Mito
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(神奈川県横浜市)
  • Year and Date: 2011-12-16
• [Presentation] Ancestral developmental mechanisms in insects revealed by R NAi analysis of cricket genes [Symp osium : RNA interference- comparativ e studies of gene functions in inve rtebrates (2011)
  • Author(s): Taro Mito
  • Organizer: 8th International Congre ss on Comparative Physiology and Bi ochemistry
  • Place of Presentation: 名古屋国際会議場(愛知県名古屋市)
  • Year and Date: 2011-06-01
• [Remarks] Natureダイジェスト,2014年3月25日,18-21ページ「ゲノム編集技術CRISPR法が世界を変える」.
• [Remarks] 読売新聞,2012年9月3日,17ページ「白いコオロギ,体色黒くする遺伝子操作」.
• [Remarks] 日経バイオテクONLINE 20128月22日,「ゲノム編集技術でコオロギの標的遺伝子破壊、徳島大と広島大がNature姉妹誌に発表」.
• [Remarks] 朝日新聞,2012年8月22日,3ページ「遺伝子操作残らぬ痕跡」.
• [Remarks] 中国新聞Online,2012年8月22日,「人工酵素でコオロギ突然変異」.