| Project/Area Number |
17590149
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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 | Chiba University |
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Principal Investigator |
MAEKAWA Mamiko Chiba University, Graduate School of Medicine, Assistant, 大学院医学研究院, 助手 (20181571)
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| Co-Investigator(Kenkyū-buntansha) |
ITO Chizuru Chiba University, Graduate School of Medicine, Assistant, 大学院医学研究院, 助手 (80347054)
TOYAMA Yoshiro Chiba University, Graduate School of Medicine, Lecturer, 大学院医学研究院, 講師 (70009637)
SUZUKI Fumie (TOYOTA Fumie) Chiba University, Graduate School of Medicine, Associate Professor, 大学院医学研究院, 助教授 (60009751)
TOSHIMORI Kiyotaka Chiba University, Graduate School of Medicine, Professor, 大学院医学研究院, 教授 (20094097)
KADOMATSU Kenji Nagoya University, Graduate School of Medicine, Professor, 大学院医学系研究科, 教授 (80204519)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2005: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | basigin / spermatogenesis / sperm maturation / fertilization / immunoglobulin superfamily / testis / sperm / glycoprotein / Basigin |
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| Research Abstract |
1. Antibody B5 was produced against the polypetide coded in exon 5, which is considered to be important during sperm maturation. In addition, commercial antibodies G19 and M190, which recognize C-terminal and N-terminal of basigin, respectively, were purchased from Santa Cruz Biotechnology.
2. Using these three antibodies, localization of basigin in the mouse testis and epididymis were studied. Sperm from cauda epididimidis, induced acrosome reaction by A23187, were also examined. Among these three antibodies, G19 showed most intensive reaction compared to M190 and B5. Further, the immunoreactivities revealed considerable differences among antibodies. For example, M190 and B5 recognized the principal segment of acrosome of the sperm head, in contrast, G19 immunostained the equatorial segment of the acrosome. It is possible that modification of the higher order structure of basigin molecule can affect the recognition of the antibodies, as well as the difference of their epitope domain.
3. Under IVF condition, basigin was detected by antibody B5 in the head of the acrosome-reacted sperm, which binds zona pellucida. It is suggested that basigin has an important role in fertilization.
4. Proteins were extracted from mouse testis and sperm from caput, corpus and cauda epididymidis and analyzed by Western blotting. Antibody M190 recognized testis-type basigin (38-40kDa) strongly rather than sperm-type basigin (26-29kDa). After PNGase F treatment and after acrosome reaction, the molecular mass of the baigin became smaller.
5. To observe the behavior of basigin protein in the testis, we made the expression construct that contains a testicular germ cell specific calmegin promoter followed by the Bsg-GFP fusion gene (cmg-Bsg-GFP). Now we generate the transgenic mice using cmg-Bsg-GFP.
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