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Analysis of function of sperm acrosomal proteins on fertilization by intra-oviductal injection

Research Project

Project/Area Number 13670020
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General anatomy (including Histology/Embryology)
Research InstitutionMiyazaki Medical College

Principal Investigator

YOSHINAGA Kazuya  Miyazaki Medical College, Anatomy, Assoc. Professor, 医学部, 助教授 (50136719)

Co-Investigator(Kenkyū-buntansha) TOSHIMORI Kiyotaka  Miyazaki Medical College, Anatomy, Professor, 医学部, 教授 (20094097)
TANII Ichiro  Miyazaki Medical College, Anatomy, Lecture, 医学部, 助手 (40207171)
Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2002: ¥500,000 (Direct Cost: ¥500,000)
KeywordsFertilization / Microinjection / Sperm / Intra-acrosomal protein / Monoclonal antibody / Egg / Sperm-egg interaction / In vitro fertilization / 先体反応 / 生殖生物学
Research Abstract

The anti-MN9 monoclonal antibody (mMN9)-containing and control solutions were injected directly into the right and left oviductal ampullae, respectively, of anesthetized female mice to assess the effect of mMN9 on fertilization in vivo. After hCG treatment, the females were mated, and their oviductal eggs and implanted embryos were examined. The rates of fertilization and of concomitant pregnancy were significantly lower than in the control side (p<0.05). In addition, histological studies showed no evidence of pathological changes in the female reproductive tract after the injections. These results indicate that mMN9 inhibits mouse fertilization significantly under in vivo conditions and that this injection method should be useful for studying the effects of antibodies and agents on fertilization in vivo. Additionally, anti-MN7, -MC41, and ?MC101 antibodies were also retained in the oviductal lumen at 20 h after injection.
We further examined interactions of MC41 with acrosomal proteases and zona proteins. A serine protease of 75 kDa was associated with MC41 under low-salt conditions, presumably forming a complex. Far Western blotting technique indicated that MC41 bound to both ZP2, and ZP2_f in the presence of high-salt- soluble sperm proteins. In acrosomal-reacting sperm, MC41 was present on the hybrid vesicles formed by the fusion of the plasma and outer acrosomal membranes. These results indicate that MC41 has a significant role in secondary sperm-zona binding during the acrosomal reaction.

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] Yoshinaga K., et al.: "Inhibition of mouse fertilization in vivo by intra-oviductal injection of an anti-equatorin monoclonal antibody"Reproduction. 122. 649-655 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tanii I., et al.: "A mouse acrosomal cortical matrix protein, MC41, has ZP2-binding activity and forms a complex with a 75-kDa serine protease"Developmental Biology. 238. 332-341 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Saxena D.K., et al.: "Are germ cell factors essential in the testicular enlargement after neonatal hypothyroidism recovery? A study using W/W^v mutant mice"International Journal of Andrology. 25. 11-18 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tanii I., Yoshinaga K.: "Change in expression of a 90 kDa-glycoprotein GP90-MC301 during prenatal and postnatal testicular development : a differentiation marker for rat germ cells"Histochemistry and Cell Biology. 118. 409-414 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] On-oka T., et al.: "The change of the activity of sperm nitric oxide synthase in the oviductal reservoir during ovulation"Reproductive Medicine and Biology. 2. 75-81 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshinaga K., Toshimori K.: "Organization and modifications of sperm acrosomal molecules during sper-matogenesis and epididymal maturation"Microscopy Research and Technique. 61. 39-45 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshinaga K., Saxena D. K., Oh-oka T., Tanii I. and Toshimori K.: "Inhibition of mouse fertilization in viva by intra-oviductal injection of an anti-equatorin monoclonal antibody."Reproduction. 122. 649-655 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tanii I., Oh-oka T., Yoshinaga K. and Toshimori K.: "A mouse acrosomal cortical matrix protein, MC41, has ZP2-binding activity and forms a complex with a 75kDa-serine protease."Developmental Biology. 238. 332-341 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Saxena D. K., Yoshinaga K., Tanii I. and Toshimori K.: "Are germ cell factors essential in the testicular enlargement after neonatal hypothyroidism recovery? A study using W/W^v mutant mice model."International Journal of Andrology. 25. 11-18 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tanii I. and Yoshinaga K.: "Change in expression of a 90kDa-glycoprotein GP90-MC301 during prenatal and postnatal testicular development : a differentiation marker for rat germ cells."Histochemistry and Cell Biology. 118. 409-414 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Oh-oka T., Saxena D. K., Tanii I., Yoshinaga K. and Toshimori K.: "The change of the activity of sperm nitric oxide synthase in the oviductal reservoir during ovulation."Reproductive Medicine and Biology. 2. 75-81 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshinaga K. and Toshimori K.: "Organization and modifications of sperm acrosomal molecules during spermatogenesis and epididymal maturation."Microscopy Research and Technique. 61. 39-45 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshinaga K., et al.: "Effects of monoclonal antibodies against sperm acrosomal proteins on mouse fertilization in vivo by intra-oviductal injection"Acta Anatomica Nipponica. 77(Suppl.). 43 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Saxena D.K., et al.: "Are germ cell factors essential in the testicular enlargement after neonatal hypothyroidism recovery? A study using W/W^V mutant mice model"International Journal of Andrology. 25. 11-18 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Tanii I., Yoshinaga K.: "Change in expression of a 9OkDa-glycoprotein GP9O-MC301 during prenatal and postnatal testicular development : a differentiation marker for rat germ cells"Histochemistry and Cell Biology. 118. 409-414 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Yoshinaga K., et al.: "Immunolocalization of acrosomal matrix protein MC41 in hamster sperm during fertilization"Asian Journal of Andrology. 43(Suppl.). 109 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Oh-oka T., et al.: "The change of the activity of sperm nitric oxide synthase in the oviductal reservoir during ovulation"Reproductive Medicine and Biology. 2. 75-81 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Yoshinaga K., Toshimori K.: "Organization and modifications of sperm acrosomal molecules during spermatogenesis and epididymal maturation"Microscopy Research and Technique. (In press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Yoshinaga K., et al.: "Inhibition of mouse fertilization in vivo by intra-oviductal injection of an anti-equatorin monoclonal antibody"Reproduction. 122. 649-655 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Yoshinaga K., et al.: "Effect of the enti-equatorin antibody mMN9 on mouse fertilization in vivo by intra-oviductal injection"Development Growth & Differentiation. 43(Suppl.). s120 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Yoshinaga K., Toshimori K.: "Organization and modifications of sperm acrosomal molecules during spermatogenesis and epididymal maturation"Microscopy Research & Technique. (in press). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Tanii I., et al.: "A mouse acrosomal cortical matrix protein, MC41, has ZP2-binding activity and forms a complex with a 75-kDa serine protease"Developmental Biology. 238. 332-341 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Tanii I., et al.: "Complex of a mouse acrosomal cortical matrix protein, MC41, and a 75kDa-serine protease shows ZP2-binding activity"Development Growth & Differentiation. 43(Suppl.). S121 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Toshimori K.: "Sperm-egg fusion in mammals. In : Andrology in the 21^<st> Century. (eds. by Robaire B, Chemes H. Morales C.R.)"MEDIMOND Inc.. 547(247-257) (2001)

    • Related Report
      2001 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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