| Project/Area Number |
09671714
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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 |
Obstetrics and gynecology
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| Research Institution | Keio University |
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Principal Investigator |
SUEOKA Kou Keio Univ., School of Medicine, Assistant Professor, 医学部, 専任講師 (90162833)
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| Co-Investigator(Kenkyū-buntansha) |
TSUCHIYA Shinichi Keio Univ., School of Medicine, Instructor, 医学部, 助手 (70276327)
SHINOHARA Masami Keio Univ., School of Medicine, Instructor, 医学部, 助手 (10276321)
YOSHIMURA Yasunori Keio Univ., School of Medicine, Professor, 医学部, 教授 (10129736)
小林 紀子 慶應義塾大学, 医学部, 助手 (50276325)
黒島 正子 慶應義塾大学, 医学部, 助手 (20255508)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1998: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1997: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Cell-free decondensing system / disulfide bond / lysolecithin / dithioshreitol / histon H2B / protamine / tapering sperm / nucleoplasmin / cell-free脱凝縮 / Triton X-100 / Xenopus laevis / protamine |
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| Research Abstract |
Sperm nuclear proteins were studied to reveal the process of human sperm decondensation. Using in vitro decondensing system sperm nuclear transformation were clearly observed. This decondensing system was 0.05% lysolecithin - 5mM dithiothreitol - 1.OM NaCl solution. Most of sperm nuclei were decondensed completely in this system. Basic nuclear proteins were extracted from decondensed nuclei with 35% acetic acid or O.4N H_2S0_4, and were separated on SDS-PRGE and transferred onto a nitrocellulose membrane. Following incubation with anti-human histone H2B antibody and HRP-labelled secondary antibody, a blot was observed. This blot exhibited mobilities of 66 kDa. Furthermore another experiment showed the existence of histone H2B.Using anti- histone H2B antibody and FITC-labelled secondary antibody, the morphological and fluorescent changes of gradually decondensing sperm nuclei were observed on a glass slide. These results suggested that human sperm nuclei contain histones considerably after nucleoproteins were replaced by protamines in the process of sperm nuclear maturation.
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