Molecular mechanism of gamete fusion in mammals

2013 Fiscal Year Final Research Report

• Project Area: Elucidation of Common Mechanisms for Allogeneic Authentication in Animals and Plants
• Project/Area Number: 21112006
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: National Research Institute for Child Health and Development
• Principal Investigator: MIYADO KENJI 独立行政法人国立成育医療研究センター, 生殖・細胞医療研究部, 室長 (60324844)
• Co-Investigator(Kenkyū-buntansha): INOUE Naokazu 福島県立医科大学, 医学部附属生体情報伝達研究所・細胞科学研究部門, 准教授 (50379096)
• Project Period (FY): 2009-07-23 – 2014-03-31
• Keywords: 膜融合 / 哺乳類 / CD9 / IZUMO1 / エキソソーム

Research Abstract

In mice, two membrane proteins, CD9 and IZUMO1, are required for sperm-egg fusion. Moreover, CD9-containing vesicles, termed egg exosomes, are released from eggs and transfered to the sperm, and then facilitate sperm fusion with the egg. Here we investigated that the egg exosomes play a role in conferring the nature of apoptotic cells on the sperm membrane by transferring inside-out CD9. Our results provide an insight into fertilization, proposing the sperm-egg fusion is controlled by a mechanism shared with phagocytosis of apoptotic cells. Moreover, we dissected IZUMO1 protein to determine the domains that were required for the function of sperm-egg fusion. We found that a fragment of the N terminus (Asp5 to Leu113) interacts with fertilization inhibitory antibodies. It also binds to the egg surface and effectively inhibits fusion in vitro. These findings suggest that the formation of a helical dimer at the N-terminal region of IZUMO1 is required for its function.

Research Products

• [Journal Article] Absence of CD9 reduces endometrial VEGF secretion and impairs uterine repair after parturition (2014)
  • Author(s): Kawano N, Miyado K, Yoshii N, Kanai S, Saito H, Miyado M, Inagaki N, Odawara Y, Hamatani T, Umezawa A
  • Journal Title: Sci Rep Volume: 4 Pages: 4701
  • DOI: 10.1038/srep04701
• [Journal Article] 精子と卵子の膜融合.受精メカニズム新論争 (2014)
  • Author(s): 井上直和、宮戸健二
  • Journal Title: 細胞工学 Volume: Vol.33. No.4
• [Journal Article] Derivation of human decidua-like cells from amnion and menstrual blood (2014)
  • Author(s): Sugawara K, Hamatani T, Yamada M, Ogawa S, Kamijo S, Kuji N, Akutsu H, Miyado K, Yoshimura Y, Umezawa A
  • Journal Title: Sci Rep Volume: 4 Pages: 4599
  • DOI: 10.1038/srep04599
• [Journal Article] Seminal vesicle protein SVS2 is required for sperm survival in the uterus (2014)
  • Author(s): Kawano N, Araki N, Yoshida K, Hibino T, Ohnami N, Makino M, Kanai S, Hasuwa H, Yoshida M, Miyado K, Umezawa A
  • Journal Title: Proc Natl Acad Sci U S A Volume: 111(11) Pages: 4145-4150
  • DOI: 10.1073/pnas.1320715111
• [Journal Article] Age-associated telomere shortening in mouse oocytes (2013)
  • Author(s): Yamada-Fukunaga T, Yamada M, Hamatani T, Chikazawa N, Ogawa S, Akutsu H, Miura T, Miyado K, Tarin JJ, Kuji N, Umezawa A, Yoshimura Y
  • Journal Title: Reprod Biol Endocrinol Volume: 11 Pages: 108
  • DOI: 10.1186/1477-7827-11-108
• [Journal Article] Molecular dissection of IZUMO1, a sperm protein essential for sperm-egg fusion (2013)
  • Author(s): Inoue N, Hamada D, Kamikubo H, Hirata K, Kataoka M, Yamamoto M, Ikawa M, Okabe M, Hagihara Y
  • Journal Title: Development Volume: 140(15) Pages: 3221-3229
  • DOI: 10.1242/dev.094854
• [Journal Article] β-catenin functions pleiotropically in differentiation and tumorigenesis in mouse embryo-derived stem cells (2013)
  • Author(s): Okumura N, Akutsu H, Sugawara T, Miura T, Takezawa Y, Hosoda A, Yoshida K, Ichida JK, Yamada M, Hamatani T, Kuji N, Miyado K, Yoshimura Y, Umezawa A
  • Journal Title: PLoS One Volume: 8(5) Pages: e63265
  • DOI: 10.1371/journal.pone.0063265
• [Journal Article] Integration of the mouse sperm fertilization-related protein equatorin into the acrosome during spermatogenesis as revealed by super-resolution and immunoelectron microscopy (2013)
  • Author(s): Ito C, Yamatoya K, Yoshida K, Fujimura L, Hatano M, Miyado K, Toshimori K
  • Journal Title: Cell Tissue Res Volume: 352(3) Pages: 739-750
  • DOI: 10.1007/s00441-013-1605-y
• [Journal Article] Production of mouse pups from germline transmission-failed knockout chimeras (2013)
  • Author(s): Fujihara Y, Kaseda K, Inoue N, Ikawa M, Okabe M
  • Journal Title: Transgenic Res Volume: 22(1) Pages: 195-200
  • DOI: 10.1007/s11248-012-9635-x
• [Journal Article] テトラスパニンによる受精の膜融合制御 (2013)
  • Author(s): 宮戸健二
  • Journal Title: 医学のあゆみ Volume: Vol.244, No.2 Pages: 181
• [Journal Article] SPACA1-deficient male mice are infertile with abnormally shaped sperm heads reminiscent of globozoospermia (2012)
  • Author(s): Fujihara Y, Satouh Y, Inoue N, Isotani A, Ikawa M, Okabe M
  • Journal Title: Development Volume: 139(19) Pages: 3583-3589
  • DOI: 10.1242/dev.081778
• [Journal Article] Visualization of the moment of mouse sperm-egg fusion and dynamic localization of IZUMO1 (2012)
  • Author(s): Satouh Y, Inoue N, Ikawa M, Okabe M
  • Journal Title: J Cell Sci Volume: 125(Pt 21) Pages: 4985-4990
  • DOI: 10.1242/jcs.100867.Epub2012Sep3
• [Journal Article] Tetraspanin-interacting protein IGSF8 is dispensable for mouse fertility (2012)
  • Author(s): Inoue N, Nishikawa T, Ikawa M, Okabe M
  • Journal Title: Fertil Steril Volume: 98(2) Pages: 465-470
  • DOI: 10.1016/j.fertnstert.2012.04.029
• [Journal Article] CD81 and CD9 work independently as extracellular components upon fusion of sperm and oocyt (2012)
  • Author(s): Ohnami N, Nakamura A, Miyado M, Sato M, Kawano N, Yoshida K, Harada Y, Takezawa Y, Kanai S, Ono C, Takahashi Y, Kimura K, Shida T, Miyado K, Umezawa A
  • Journal Title: Biol Open Volume: 1(7) Pages: 640-647
  • DOI: 10.1242/bio.20121420
• [Journal Article] Mamld1 deficiency significantly reduces mRNA expression levels of multiple genes expressed in mouse fetal Leydig cells but permits normal genital and reproductive development (2012)
  • Author(s): Miyado M, Nakamura M, Miyado K, Morohashi K, Sano S, Nagata E, Fukami M, Ogata T
  • Journal Title: Endocrinology Volume: 153(12) Pages: 6033-6040
  • DOI: 10.1210/en.2012-1324
• [Journal Article] Impaired fertilizing ability of superoxide dismutase 1-deficient mouse sperm during in vitro fertilization (2012)
  • Author(s): Tsunoda S, Kawano N, Miyado K, Kimura N, Fujii J
  • Journal Title: Biol Reprod Volume: 87(5) Pages: 121
  • DOI: 10.1095/biolreprod.112.102129
• [Journal Article] 卵由来エキソソームによる細胞融合の促進機構 (2012)
  • Author(s): 宮戸健二
  • Journal Title: 医学のあゆみ Volume: Vol.241,No.11 Pages: 861-862
• [Journal Article] Acrosome-reacted mouse spermatozoa recovered from the perivitelline space can fertilize other eggs (2011)
  • Author(s): Inoue N, Satouh Y, Ikawa M, Okabe M, Yanagimachi R
  • Journal Title: Proc Natl Acad Sci USA Volume: 108(50) Pages: 20008-20011
  • DOI: 10.1073/pnas.1116965108
• [Journal Article] β-catenin is a molecular switch that regulates transition of cell-cell adhesion to fusion (2011)
  • Author(s): Takezawa Y, Yoshida K, Miyado K, Sato M, Nakamura A, Kawano N, Sakakibara K, Kondo T, Harada Y, Ohnami N, Kanai S, Miyado M, Saito H, Takahashi Y, Akutsu H, Umezawa A
  • Journal Title: Sci Rep Volume: 1 Pages: 68
  • DOI: 10.1038/srep00068
• [Journal Article] GSTT1 is upregulated by oxidative stress through p38-MK2 signaling pathway in human granulosa cells : possible association with mitochondrial activity (2011)
  • Author(s): Ito M, Imai M, Muraki M, Miyado K, Qin J, Kyuwa S, Yoshikawa Y, Hosoi Y, Saito H, Takahashi Y
  • Journal Title: Aging (Albany NY) Volume: 3(12) Pages: 1213-1223
• [Journal Article] Lipid rafts : keys to sperm maturation, fertilization, and early embryogenesis (2011)
  • Author(s): Kawano N, Yoshida K, Miyado K, Yoshida M
  • Journal Title: J Lipids Volume: 2011 Pages: 264706
  • DOI: 10.1155/2011/264706
• [Journal Article] Lipid rafts enriched in monosialylGb5Cer carrying the stage-specific embryonic antigen-4 epitope are involved in development of mouse preimplantation embryos at cleavage stage (2011)
  • Author(s): Sato B, Katagiri YU, Miyado K, Okino N, Ito M, Akutsu H, Okita H, Umezawa A, Fujimoto J, Toshimori K, Kiyokawa N
  • Journal Title: BMC Dev Biol Volume: 11 Pages: 22
  • DOI: 10.1186/1471-213X-11-22
• [Journal Article] Calsperin is a testis-specific chaperone required for sperm fertility (2011)
  • Author(s): Ikawa M, Tokuhiro K, Yamaguchi R, Benham AM, Tamura T, Wada I, Satouh Y, Inoue N, Okabe M
  • Journal Title: J Biol Chem Volume: 286(7) Pages: 5639-56465639-5646
  • DOI: 10.1074/jbc.M110.140152
• [Journal Article] Identification and disruption of sperm-specific angiotensin converting enzyme-3 (ACE3) in mouse (2010)
  • Author(s): Inoue N, Kasahara T, Ikawa M, Okabe M
  • Journal Title: PLoS One Volume: 5(4) Pages: e10301
  • DOI: 10.1371/journal.pone.0010301
• [Journal Article] Sperm equatorial segment protein 1, SPESP1, is required for fully fertile sperm in mouse (2010)
  • Author(s): Fujihara Y, Murakami M, Inoue N, Satouh Y, Kaseda K, Ikawa M, Okabe M
  • Journal Title: J Cell Sci Volume: 123(Pt 9) Pages: 1531-1536
  • DOI: 10.1242/jcs.067363
• [Journal Article] Fertilization : a sperm's journey to and interaction with the oocyte (2010)
  • Author(s): Ikawa M, Inoue N, Benham AM, Okabe M
  • Journal Title: J Clin Invest Volume: 120(4) Pages: 984-994
  • DOI: 10.1172/JCI41585
• [Journal Article] Age-associated changes in the subcellular localization of phosphorylated p38 MAPK in human granulosa cells (2010)
  • Author(s): Ito M, Miyado K, Nakagawa K, Muraki M, Imai M, Yamakawa N, Qin J, Hosoi Y, Saito H, Takahashi Y
  • Journal Title: Mol Hum Reprod Volume: 16(12) Pages: 928-937
  • DOI: 10.1093/molehr/gaq076
• [Journal Article] Mice lacking two sperm serine proteases, ACR and PRSS21, are subfertile, but the mutant sperm are infertile in vitro (2010)
  • Author(s): Kawano N, Kang W, Yamashita M, Koga Y, Yamazaki T, Hata T, Miyado K, Baba T
  • Journal Title: Biol Reprod Volume: 83(3) Pages: 359-369
  • DOI: 10.1095/biolreprod.109.083089
• [Journal Article] Tetraspanin family protein CD9 in the mouse sperm : unique localization, appearance, behavior and fate during fertilization (2010)
  • Author(s): Ito C, Yamatoya K, Yoshida K, Maekawa M, Miyado K, Toshimori K
  • Journal Title: Cell Tissue Res Volume: 340(3) Pages: 583-594
  • DOI: 10.1007/s00441-010-0967-7
• [Journal Article] Equatorin : identification and characterization of the epitope of the MN9 antibody in the mouse (2009)
  • Author(s): Yamatoya K, Yoshida K, Ito C, Maekawa M, Yanagida M, Takamori K, Ogawa H, Araki Y, Miyado K, Toyama Y, Toshimori K
  • Journal Title: Biol Reprod Volume: 81(5) Pages: 889-897
  • DOI: 10.1095/biolreprod.109.077438
• [Presentation] テトラスパニンを介した細胞融合の分子メカニズム (2013)
  • Author(s): 宮戸健二
  • Organizer: ワークショップ「4回膜貫通蛋白質の構造と機能解析の進展」、第36回日本分子生物学会年会
  • Year and Date: 20130000
• [Presentation] 受精の膜融合におけるCD9およびエキソソームの役割 (2012)
  • Author(s): 宮戸健二
  • Organizer: シンポジウム「バイオイメージングにより明らかにされた動・植物の有性生殖メカニズム」,日本顕微鏡学会第68回学術講演会
  • Year and Date: 20120000
• [Presentation] The mechanism of sperm-egg fusion in mouse and the involvement of IZUMO1 and a novel factor (2011)
  • Author(s): Inoue, N
  • Organizer: Gordon Research Conferences "Cell-Cell Fusion In Sex, Life, Development and Disease"
  • Place of Presentation: University of New England (Biddeford, U. S. A.)
  • Year and Date: 2011-08-09
• [Presentation] CD9 action in the egg and on the sperm of mice (2009)
  • Author(s): Miyado, K
  • Organizer: Cell-Cell Fusion, Gordon Research Conferences
  • Place of Presentation: Colby-Sawyer College (New London, U.S.A.)
  • Year and Date: 2009-07-20
• [Book] Sexual Reproduction in Animals and Plants (edited by Sawada H, Inoue N, Iwano M) (2014)
  • Publisher: Springer
• [Book] 動植物の受精学 - 共通性と多様性 - (澤田均 編) (2014)
  • Publisher: 化学同人
• [Book] Role of CD9 in sperm-egg fusion and its general role in fusion phenomena. Cell Fusions (edited by Lars-Inge Larsson) (2011)
  • Author(s): Kawano N, Harada Y, Yoshida K, Miyado K
  • Publisher: Springer
• [Remarks]
  • URL: http://www.ncchd.go.jp/research/nch/reproduction/staff_miyado.html