Intercellular recognition between sperm and eggs: Studies on the allogeneic and xenogeneic molecular recognition

• Project/Area Number: 17H03672
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Cell biology
• Research Institution: Kinjo Gakuin University (2019-2020); Nagoya University (2017-2018)
• Principal Investigator: Sawada Hitoshi 金城学院大学, 生活環境学部, 教授 (60158946)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2019: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2018: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2017: ¥7,670,000 (Direct Cost: ¥5,900,000、Indirect Cost: ¥1,770,000)
• Keywords: 受精 / 自家不和合性 / 細胞間相互作用 / ホヤ / 精子 / 卵 / ライシン / プロテアーゼ / 昆虫細胞 / 自己非自己認識

Outline of Final Research Achievements

Ascidians are hermaphrodites, which release sperm and eggs nearly simultaneously, but self-fertilization is prohibited in Ciona intestinalis and Halocynthia roretzi. In C. intestinalis, we reported that two gene pairs, s-Themis-A and v-Themis-A in locus A, and s-Themis-B and v-Themis-B in locus B, play a pivotal role in self-sterility. Here, we identified one addition gene pair s/v-Themis-B2 in locus B involved in this system. Therefore, three multi-allelic gene pairs, s/v-Themis-A, -B, and B2, are responsible for self-sterility in C. intestinalis. We previously showed that the increase in intracellular Ca2+ takes place after sperm recognizes the vitelline coat of the egg as self-egg, resulting in the detachment from the vitelline coat. Here, we revealed low Ca2+ seawater enables self-fertilization, probably because of inhibition of Ca2+. However, these phenomena were never observed in H. roretzi, suggesting that the mechanism of self-sterility appears to be different.

Academic Significance and Societal Importance of the Research Achievements

ホヤの自家不稔性機構は動植物共通の生殖原理に基づくと考えられる。また、カタユウレイボヤで発見された精子s-Themisは、哺乳類精子のPKDREJと相同な分子で、多型がみられる。従って、配偶子間相互作用において同系統個体と認識されて受精が阻害され不妊になる可能性も考えられる。ホヤの受精研究から、ヒトの不妊診断・治療の研究に発展する可能性も考えられ、社会的にも臨床医学的にも意義深い研究である。

Research Products

• [Journal Article] A dynein-associated photoreceptor protein prevents ciliary acclimation to blue light (2021)
  • Author(s): Kutomi O, Yamamoto R, Hirose K, Mizuno K, Nakagiri Y, , , , , , , Shibata D, Shibata M, Shiba K, Kita M, Kigoshi H, Tanaka Y, Yamasaki Y, Asahina Y, Song C, Nomura M, Nomura M, Nakajima A, Nakachi M, Yamada L, Nakazawa S, Sawada H, Murata K, Mitsuoka K, Ishikawa T, Wakabayashi K, Kon T, Inaba K
  • Journal Title: Science Advances Volume: 7 Issue: 9
  • DOI: 10.1126/sciadv.abf3621
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Phylogenetic comparisons reveal mosaic histories of larval and adult shell matrix protein deployment in pteriomorph bivalves (2020)
  • Author(s): Ran Zhao, Takeshi Takeuchi, Ryo Koyanagi, Alejandro Villar-Briones, Lixy Yamada, Hitoshi Sawada, Akito Ishikawa, Shunsuke Iwanaga, Kiyohito Nagai, Yuqi Che, Noriyuki Satoh, Kazuyoshi Endo
  • Journal Title: Scientific Reports Volume: 10(1) Issue: 1 Pages: 22140-22140
  • DOI: 10.1038/s41598-020-79330-x
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Vitelline membrane proteins promote left-sided nodal expression after neurula rotation in the ascidian, Halocynthia roretzi. (2019)
  • Author(s): Tanaka, Y., Yamada, S., Connop, S. L., Hashii, N., Sawada, H., Shih, Y., and Nishida, H.
  • Journal Title: Dev. Biol. Volume: 449 Issue: 1 Pages: 52-61
  • DOI: 10.1016/j.ydbio.2019.01.016
  • Peer Reviewed
• [Journal Article] Peanut agglutinin specifically binds to a sperm region between the nucleus and mitochondria in tunicates and sea urchins. (2018)
  • Author(s): Nakazawa S, Shirae-Kurabayashi M, Sawada H.
  • Journal Title: Molecular Reproduction and Development Volume: 印刷中 Issue: 6 Pages: 464-477
  • DOI: 10.1002/mrd.22982
  • Peer Reviewed
• [Journal Article] Chitin-based barrier immunity and Its loss predated mucus-colonization by indigenous gut microbiota. (2018)
  • Author(s): Nakashima, K., Kimura, S., Ogawa, Y., Watanabe, S., Soma, S., Kaneko, T., Yamada, L., Sawada, H., Tung, C.-H., Lu, T-M., Yu, J-K., Briones, A. V., Kikuchi, S., and Satoh, N.
  • Journal Title: Nature Communications Volume: 9(1) Issue: 1 Pages: 3402-3402
  • DOI: 10.1038/s41467-018-05884-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The male gamete membrane protein DMP9/DAU2 is required for double fertilization in flowering plants. (2018)
  • Author(s): Takahashi T., Mori, T., Ueda, K., Yamada, L., Nagahara, S., Higashiyama, T., Sawada, H., Igawa, T.
  • Journal Title: Development Volume: 145(23) Issue: 23
  • DOI: 10.1242/dev.170076
  • Peer Reviewed
• [Journal Article] Dual gene repertories for larval and adult shells reveal molecules for molluscan shell formation. (2018)
  • Author(s): Zhao, R., Takeuchi, T., Luo, Y-J., Ishikawa, A., Kobayashi, T., Koyanagi, R., Villar-Briones, A., Yamada, L., Sawada, H., Iwanaga, S., Nagai, K., Satoh, N., and Endo, K.
  • Journal Title: Molecular Biology and Evolution, Volume: 35(11) Pages: 2751-2761
  • DOI: 10.1093/molbev/msy172
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The role of metalloproteases in fertilisation in the ascidian Ciona robusta. (2018)
  • Author(s): Nakazawa, S., Shirae-Kurabayashi, M., and Sawada, H.
  • Journal Title: Scientific Reports, Volume: 9(1) Issue: 1 Pages: 1009-1009
  • DOI: 10.1038/s41598-018-37721-1
  • Peer Reviewed / Open Access
• [Journal Article] Ascidian sexual reproductive strategies: Mechanisms of sperm-egg interaction and self-sterility. (2018)
  • Author(s): Sawada H., Nakazawa, S., and Shirai-Kurabayashi, M.
  • Journal Title: Reproductive and Developmental Strategies: Diversity and Commonality in Animal (Springer Japan) Volume: 印刷中
  • Peer Reviewed
• [Journal Article] 受精 (2018)
  • Author(s): 澤田 均
  • Journal Title: 動物の百科辞典 Volume: 印刷中
  • Peer Reviewed
• [Journal Article] Haemorrhagic snake venom metalloproteases and human ADAMs cleave LRP5/6, which disrupts cell–cell adhesions in vitro and induces haemorrhage in vivo (2017)
  • Author(s): Seo T, Sakon T, Nakazawa S, Nishioka A, Watanabe K, Matsumoto K, Akasaka M, Shioi N, Sawada H, Araki S
  • Journal Title: FEBS Journal Volume: 284 Issue: 11 Pages: 1657
  • DOI: 10.1111/febs.14066
  • Peer Reviewed / Open Access
• [Journal Article] Spawning of the ascidian Halocynthia roretzi. (2017)
  • Author(s): Sawada, H., Shirae-Kurabayashi M., and Takaharu Numakunai
  • Journal Title: Molecular Reproduction and Development Volume: 84 Issue: 2 Pages: 83-83
  • DOI: 10.1002/mrd.22776
  • Peer Reviewed / Open Access
• [Journal Article] Isolation of GFP-tagged plasma membrane protein from Arabidopsis egg cells (2017)
  • Author(s): Igawa, T., Yamada, L., Sawada, H., Mori, T.
  • Journal Title: Plant Biotechnology Volume: 34 Issue: 2 Pages: 119-123
  • DOI: 10.5511/plantbiotechnology.17.0522a
  • NAID: 130006789291
  • ISSN: 1342-4580, 1347-6114
  • Peer Reviewed / Open Access
• [Presentation] ホヤの自家不和合機構と精子プロテアーゼの役割 (2020)
  • Author(s): 澤田 均
  • Organizer: 第5回ホヤ研究会
• [Presentation] ホヤの受精研究から学んだこと (2020)
  • Author(s): 澤田 均
  • Organizer: 第7回生殖若手の会
  • Invited
• [Presentation] ホヤの受精機構:自家不和合性とライシンについて (2018)
  • Author(s): 澤田均
  • Organizer: 日本動物学会第89回大会
  • Invited
• [Presentation] 昆虫細胞を用いたカタユウレイボヤs-Themis, v-Themisタンパク質の相互作用解析 (2018)
  • Author(s): 成山 奏子、白江-倉林 麻貴、澤田 均
  • Organizer: 日本動物学会第89回大会
• [Presentation] ADAM-LRP信号伝達経路に関わる新たな因子 (2018)
  • Author(s): 酒井健有、澤田均、荒木聡彦
  • Organizer: 日本動物学会中部支部大会
• [Presentation] 昆虫細胞を用いたカタユウレイボヤs-Themis、v-Themisタンパク質の相互作用についての研究 (2018)
  • Author(s): 成山 奏子 、西尾 俊亮 、白江-倉林 麻貴、松田 幹 、澤田 均
  • Organizer: 日本動物学会中部支部大会
• [Presentation] A method for isolation of specific Arabidopsis egg cell membrane protein without gamete isolation procedure. (2017)
  • Author(s): Tomoko Igawa, Lixy Yamada, Hitoshi Sawada, Toshiyuki Mori
  • Organizer: Cold Spring Harbor Asia Conference on Plant Cell & Development Biology
• [Presentation] Study on molecular players regulating male and female gamete interaction during double fertilization. (2017)
  • Author(s): Taro Takahashi, Toshiyuki Mori, Lixy Yamada, Hitoshi Sawada, Kenji Ueda, Shiori Nagahara, Tetsuya Higashiyama, Tomoko Igawa
  • Organizer: 第59回日本植物学会
• [Presentation] TALENを用いたカタユウレイボヤにおける精子タンパク質Urabinの機能解析 (2017)
  • Author(s): 成山奏子、白江-倉林麻貴、中澤志織、澤田均
  • Organizer: 第88回日本動物学会
• [Presentation] Roles of ascidian sperm metalloproteases in fertilization of a marine invertebrate Ciona robusta (2017)
  • Author(s): Shiori Nakazawa, Maki Shirae-Kurabayashi, Hitoshi Sawada
  • Organizer: Gordon Research Conference: Fertilization & Activation of Development
• [Presentation] cDNA cloning and expression of two ubiquitin-activating enzymes UBA1 and UBA6 in gametes of the ascidian Halocynthia rotezi (2017)
  • Author(s): Sawada, H., Inoue, S., Otsuka, K., Shirae-Kurabayashi
  • Organizer: Gordon Research Conference: Fertilization & Activation of Development
• [Presentation] Vascular endothelial cell-cell adhesion disruption and hemorrhage are induced by truncated LRP5/6 which is processed by hemorrhagic snake venom metalloproteases and human ADAMs (2017)
  • Author(s): Seo, T., Kakon, T., Nakazawa, S., Nishioka, A., Watanabe, K., Matsumoto, K., Akasaka, M., Shioi, N., Sawada, H., and Araki, S.
  • Organizer: Gordon Research Conferences
• [Book] Reproductive Biology and Aquaculture Technology in Aquatic Animals (Eds. Yoshida, M., Asturiano J. F.) (Chapter 9; Self- and nonself-recognition of gametes in ascidians.) (2019)
  • Author(s): Sawada, H. and Shirae-Kurabayashi, M.
  • Publisher: Springer
• [Book] Reproductive and Developmental Strategies: The Continuity of Life (Y. Iwao, S. Kobayashi eds.) (Chapter 23 Ascidian sexual reproductive strategies: Mechanisms of sperm-egg interaction and self-sterility.) (2018)
  • Author(s): Sawada H., Nakazawa, S., and Shirai-Kurabayashi, M ほか72人
  • Total Pages: 789
  • Publisher: Springer
  • ISBN: 9784431566076
• [Book] 動物学の百科事典 (2018)
  • Author(s): 公益社団法人日本動物学会
  • Total Pages: 800
  • Publisher: 丸善出版
  • ISBN: 4621303090