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MOLECULAR CYTOCHEMICAL ANALYSIS OF GERM CELL DEATH IN SPERMATOGENESIS.

Research Project

Project/Area Number 07680771
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Cell biology
Research InstitutionNagasaki University

Principal Investigator

KOJI Takehiko  Nagasaki University, School of Medicine, Associate professor., 医学部, 助教授 (30170179)

Co-Investigator(Kenkyū-buntansha) NAKANE Kazuo  Nagasaki University, School of Medicine, Professor., 医学部, 教授 (60164240)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1996: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1995: ¥1,000,000 (Direct Cost: ¥1,000,000)
Keywordsspermatogenesis / apoptosis / necrosis / Fas / Fas ligand / molecular histochemistry / mouse / アポトーシス / ネクローシス / マウス精巣 / in situ hibridization / 免疫組織化学 / エストロゲン / 潜伏睾丸 / in situ nick tranolation / in situ TdT / lpr / gld
Research Abstract

Germ cell death is very common in mammalian testes during developmental as well as adult period. In mouse testis, it was known that a small, but significant number of germ cells are spontaneously degenerated during adult life and in fact 25-75% of the expected sperm yield are thought to be lost in the mature spermatogenesis. Also, in artificial cryptorchidism and estrogen-treated (i.p.injection of 30 mu g beta-estradiol-3-benzoate/head, every 5 day for 60 days) mice, the degeneration of germ cells are much more prominent. However, we are not sure wether the germ cell death is of apoptotic nature or necrotic nature, and moreover our knowledge on the underlying mechanism to induce the germ cell death is only limited. Therefore, in this study, we attempted to clarify the type of cell death, which occurs in normal as well as the damaged mouse testes, by both electron microscopy and in situ nick translation and/or TUNEL.As a result, we found that most of the degenerating germ cells were of apoptotic nature in normal and damaged testes, and that in the normal testis apoptosis of spermatogonia and spermatocytes was most abundant in the stage XII of seminiferous epithelium cycle. To elucidate the induction mechanism of germ cell apoptosis, we investigated the spatial and temporal relationship of the expression of Fas/Fas ligand with the appearance of apoptosis molecular-histochemically. Then we did not find any positive correlation between them in normal testis, whereas in estrogen-treated mouse testis both expressions of Fas and Fas ligand were markedly increased in germ cells and Sertoli cells, respectively, and seemed to be closely associated with the apoptotic germ cells. Consequently, these results indicate that the Fas/Fas ligand system may be involved in the induction of germ cell apoptosis in damaged testis, but not in normal spermatogenesis.

Report

(4 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • Research Products

    (35 results)

All Other

All Publications (35 results)

  • [Publications] Koji, T: "Recent advances in molecular histochemistry:In situ hybridization and south western histochemistry" G.Electron Microsc. 45・2. 119-127 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Koji, T: "Non-radioactive in situ nick trano lation:A useful molecular histochemical tool to detect single-stranded DNA breaks." Acta Histochem.Cytochem. 29・1. 71-79 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hahuno, N: "Tias/APO-1/CD95 system as a mediator of granulosa cell apoptosis in ovarian follicle atresia." Endo crinology. 137・5. 1938-1948 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kawakami, A: "Fas and Fas legand enteraction is necessary for human osteohlast apoptosis." G.Bond and Mineral Res.12・10. 1637-1646 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Higami, Y: "Aging acceterates bid life-long dietary nestriction suppresses apoptosis-related Fas expression on hepatocytes." Am.G.Pathol.151・3. 659-663 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Wang, R.-A.: "Autonomous cell death of male germ cells during fetal and postnatal pericd" Biol.Reprod. (印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Koji, T: "Apoptosis:Its role and Mechanisim" Business Center for Academic Societies Japan (印刷中), (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Koji, T.: "Recent advances in molecular histochemistry : In situ hybridization and southwestern histochemistry." J.Electron Microsc.45(2). 119-127 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Koji, T.: "Non-radioactive in situ nick translation : A useful molecular histochemical tool to detect single-stranded DNA breaks." Acta Histochem.Cytochem.29(1). 71-79 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hakuno, N.: "Fas/APO-1/CD95 system as a mediator of granulosa cell apoptosis in ovarian follicle atresia." Endocrinology. 137(5). 1938-1948 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kawakami, A.: "Fas and Fas ligand interaction is necessary for human osteoblast apoptosis." J.Bone and Mineral Res.12(10). 1637-1646 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Higami, Y.: "Aging accelerates but life-long dietary restriction suppre sses apoptosis-related Fas expression on hepatocytes." Am.J.Pathol.151(3). 659-663 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Wang, R.-A: "Autonomous cell death of mouse male germ cells during fet al and postnatal period." Biol.Reprod.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Koji, T.: In : Apoptpsis : Its role and Mechanism.Business Center for Academic Societies Japan (in press), (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Higami, Y: "Oging accelesates but life-long dietary nestnicturn suppresses apoptones slated Fas expression on-hypatocytes." Am J. Patrol. 151・3. 659-663 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kawakami, A.: "Fas cvnd Fas ligancl interaction is necessary for human ostesblast apoptosis" J. Bone Mineral Res.12・10. 1637-1646 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Hashimoto, S.: "Freguency of apoptosis relates inversely to inwasiveness and metastatic activity in human colonetal cancer." Vischows Aoch. 431. 241-248 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Nogeae, S.: "Induction of apoptosis in ischemia reperfusion model of mouse hidgey porsilte envolvement of Fas." G. Am. Soc. Nephool.9(in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Wang, R-A.: "Autonomous cell death of mouse male germ cells during fetal and postnatal period." Biol. Reprod.58(in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Matsunami, H.: "Expression and Localization of enduceble nitruc oxide synthase in the rat ovary a possible envolvement of nitric oxide on the follicula development" Biochem. Biophys. Res. Commun. (in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Koji, T.: "Apoptosis : Its role and mechanism" Business Center for Academic Societies Japan(in press), (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Koji,T.: "Non-radicative in situ nick translation : Auseful molecular histochemical toool to detect single-stranded DNA breaks" Acta Histochem.Cytochem.29・1. 71-79 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hakuno,N: "Fas/Apo-1/CD95 system as a mediator of gramulosa cell apoptosis in ovarian follicle atresea" Endocrinology. 137.5. 1938-1948 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Iwamoto,M.: "Apoptosis of crypt epithalial cells in alcerative colitis." G.Pathol. 180. 152-159 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 小路武彦: "DNA鎖切断の分子組織細胞化学的検出法。" 電子顕微鏡. 31・2/3. 145-148 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 岩本美智子: "潰瘍性大腸炎における陰窩上皮細胞のアポトーシス。" 日本臨床. 54・7. 1970-1974 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 橋本 聡: "大腸癌組織におけるアポトーシスの検出とFasの発現。" 日本臨床. 54・7. 1912-1915 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hashimoto,S.: "Differential staining of DNA strand breaks occurring in dying cells by non-radioactive in situ nick translation" Arch.Histol.Cytol.58. 161-170 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Koji,T.: "Recent advances in molecular histochemistry : in situ hybridization and southwestern histochemistry" g.Electron.Microsc.45. in press (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Hakuno,N.: "Fas/APO-1/CD95 system as a mediator of granulosa cell apoptosis in ovarianfollicle atresia." Endocrinology. 137. in press (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Koji,T.: "Non-radioactive in situ nick translation : a useful molecular histochemical tool to detect single-stranded DNA breaks." Acta Histochem.Cytochem.29. in press (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] 小路武彦: "アポトーシスの組織細胞化学。" 日本網内系学会誌. 35. 3-9 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 小路武彦: "特集アポトーシスの細胞生物学によせて。" 細胞. 28. 2-3 (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] 小路武彦: "アポトーシス実験法(辻本賀英・刀称重信・山田武編)" 羊土社, 283 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 小路武彦: "現代病理学大系-補遺I" 中山書店, 261 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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