| Project/Area Number |
13670020
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General anatomy (including Histology/Embryology)
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| Research Institution | Miyazaki Medical College |
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Principal Investigator |
YOSHINAGA Kazuya Miyazaki Medical College, Anatomy, Assoc. Professor, 医学部, 助教授 (50136719)
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| 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)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2002: ¥500,000 (Direct Cost: ¥500,000)
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| Keywords | Fertilization / Microinjection / Sperm / Intra-acrosomal protein / Monoclonal antibody / Egg / Sperm-egg interaction / In vitro fertilization / 先体反応 / 生殖生物学 |
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| 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.
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