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A novel relaxin-like protein discovered in boar seminal vesicle : Binding on sperm plasma membrane and physiological function

Research Project

Project/Area Number 11660281
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied animal science
Research InstitutionShizuoka University

Principal Investigator

KOHSAKA Tetsuya  Shizuoka University, Faculty of Agriculture, Professor, 農学部, 教授 (10186611)

Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2000: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1999: ¥2,100,000 (Direct Cost: ¥2,100,000)
KeywordsRelaxin / Seminal vesicle / sperm / TR-FIA / Capacitation / Boar / 蛍光CTC / 精子細胞膜
Research Abstract

This research project was planned to identify the binding sites on boar sperm for relaxin-like protein, to clarify the nature of the binding sites on the sperm, and to examine possible physiological roles of the binding on the sperm. The outcome of this research is as follows.
1. Development of TR-FIA as a novel assay system : This research was successful in developing a new assay system for porcine relaxin by adapting time-resolved fluoroimmunoassay (TR-FIA) introduced recently as a non-RIA format. The assay system was a solid-phase TR-FIA based on competition for a polyclonal anti-relaxin antibody between europium (Eu) -labeled relaxin and test samples. This assay we established had several advantages : the assay had a high sensitivity, the measurement time was completed in only 5 h with one second per sample, no special safety precautions had to be taken, and the precision, reproducibility and accuracy of the assay were satisfactory.
2. Identification and nature of the binding sites on boar sperm : This research has clearly demonstrated that relaxin-like protein binds mainly to the acrosomal region of boar spermatozoa, with weaker binding at the midpiece and tail, and the binding sites are the nature of both protein and lipid, suggesting that the protein activates sperm function through a specific receptor.
3.Possible physiological roles of the binding on boar sperm : This research has shown that relaxin-like protein acts as cholesterol-docking protein and induces sperm capacitation, suggesting that the protein may have value as a potential marker for predicting sperm fertilizing ability of boars.

Report

(3 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] T.Ogine et al.: "Time-resolved fluoroimmunoassay (TR-FIA) of porcine relaxin"Experimental and Clinical Endocrinology & Diabetes 〔国際誌〕. 107. 276-280 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] J.Kobayashi et al.: "Fluorescence in situ hybridization with Y chromosome-specific probes in decondenced bovine spermatozoa"Theriogenology 〔国際誌〕. 52. 1044-1054 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] T.Kohsaka et al.: "Uterine inflammatory response after mating and in vitro detection of granulocyte-macrophage colony stimulating factor inducing potential of pig semen"Theriogenology 〔国際誌〕. 53. 475 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Y.Sasaki et al.: "Immunoreactive relaxin in seminal plasma of fertile boars and its correlation with sperm motility characteristics determined by computer-assisted digital image analysis"International Journal of Andrology 〔国際誌〕. 24. 24-30 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Ogine T, Kohsaka T, Taya K.: "Time-resolved fluoroimmunoassay (TR-FIA) of porcine relaxin.T"Experimental and Clinical Endocrinology & Diabetes. 107. 276-280 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kobayashi J, Kohsaka T, Sasada H, Umezu M, Sato E.: "Fluorescence in situ hybridization with Y chromosome-specific probes in decondenced bovine spermatozoa"Theriogenology. 52. 1044-1054 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kohsaka T, Sasada H, Hashizume K, Akita F, Kobayashi J, Kang M, Kawarasaki T, Bamba K, Takahara H.: "Uterine inflammatory response after mating and in vitro detection of granulocyte-macrophage colony stimulating factor inducing potential of pig semen"Theriogenology. 53. 475 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sasaki Y, Kohsaka T, Kawarasaki T, Sasada H, Ogine T, Bamba K, Takahara H.: "Immunoreactive relaxin in seminal plasma of fertile boars and its correlation with sperm motility characteristics determined by computer-assisted digital image analysis."International Journal of Andrology. 24. 24-30 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] T.Kohsaka et al.: "Relaxin, testosterone and fructose in seminal plasma of fertile boars and their correlation with sperm motilitv characteristics"5th Asian Symposium on Animal Biotechnology. 39 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] T.Kohsaka et al.: "In situ Hybridization of relaxin mRNA with microwave fixation and Digoxigenin-ladeled RNA probes in pregnant sow corpora lutea,"14th Inter.Congr.Animal Reproduction. 139 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] H.Sasada et al.: "Sexual performance, serum testosterone and DNA sequences related to sex determination in intersex goat"14th Inter.Congr.Animal Reproduction. 17 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Y.Sasaki et al.: "Immunoreactive relaxin in seminal plasma of fertile boars and its correlation with sperm motility characteristics determined by computer-assisted digital image analysis"International Journal of Andrology. 24・1. 24-30 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Ogine,T: "Time-resolved fluoroimmunoassay (TR-FIA) of porcine relaxin"Experimental and Clinical Endocrinology & Diabetes(国際誌). 107・4. 276-280 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kobayashi,J: "Fluorescence in situ hybrization with Y chromosome-specific probe in decondensed bovine spermatozoa"Theriogenology(国際誌). 52・6. 1044-1054 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kohsaka,T: "Uterine inflammarory response after mating and in vitro detection of granulocyte-macrophage colony stimulating factor inducing potential of pig semen"Theriogenology(国際誌). 53・1. 475 (2000)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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