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  1. 020 学位論文
  2. 生命科学研究科
  3. 18 遺伝学専攻

Characterization of type A spermatogonia and development of the culture system in zebrafish

https://ir.soken.ac.jp/records/1043
https://ir.soken.ac.jp/records/1043
3dd0eb6c-d20f-41f2-a483-6272dfeff231
名前 / ファイル ライセンス アクション
甲1161_要旨.pdf 要旨・審査要旨 (234.1 kB)
甲1161_本文.pdf 本文 (5.2 MB)
Item type 学位論文 / Thesis or Dissertation(1)
公開日 2010-02-22
タイトル
タイトル Characterization of type A spermatogonia and development of the culture system in zebrafish
タイトル
タイトル Characterization of type A spermatogonia and development of the culture system in zebrafish
言語 en
言語
言語 eng
資源タイプ
資源タイプ識別子 http://purl.org/coar/resource_type/c_46ec
資源タイプ thesis
著者名 齋藤, 憲二

× 齋藤, 憲二

齋藤, 憲二

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フリガナ サイトウ, ケンジ

× サイトウ, ケンジ

サイトウ, ケンジ

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著者 SAITO, Kenji

× SAITO, Kenji

en SAITO, Kenji

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学位授与機関
学位授与機関名 総合研究大学院大学
学位名
学位名 博士(理学)
学位記番号
内容記述タイプ Other
内容記述 総研大甲第1161号
研究科
値 生命科学研究科
専攻
値 18 遺伝学専攻
学位授与年月日
学位授与年月日 2008-03-19
学位授与年度
値 2007
要旨
内容記述タイプ Other
内容記述 Continuation of spermatogenic process throughout life relies on a proper<br />regulation of self-renewal and differentiation of spermatogonial stem cells which is<br />present in a special cellular organization called a niche. In spite of biological<br />significance of the spermatogonial stem cells, little is known about their behavior and<br />properties because of a lack of model system to approach the stem cell maintenance<br /> and differentiation in several organisms. <br />  In this study, I show the presence of subpopulations in zebrafish type A<br />spermatogonia which is classically considered homogeneous pepulation varying in<br />size, location pattern and cell cycle phase. Morphological analysis showed two types<br />of type A sperrnatogonia that are focalized distant from basement membranes (termed<br />type A<small>dt</small>) and close to basement membranes (A<small>cl</small>). The thymidine analogue <br />5-bromo-2'-deoxyuridine (BrdU) incorporation experiments showed that duration of <br />cell cycle of the type A<small>cl</small> spermatogonia was longer than that of the.type A<small>dt</small> <br />spermatogonia. Tracing label-retaining cells (LRCs), after 4 weeks of chase, I <br />observed occasionally BrdU-positive type A<small>cl</small> spermatogonia. Immunohistochemical<br /> analysis using antibody against zebrafish homologue of Synthetic lethal mutant of <br />dpb11-1 (zSld5) which is able to distinguish between the guiescent cells and the <br />proliferation cells showed that type A<small>cl</small> spermatogonia could be divided into two<br />subpopulations. One population is zSld5-positive type A<small>cl</small> spermatogonia and the other <br />is zSld5-negative type A<small>cl</small> spermatogonia In addition, many LRCs are contained in a<br />latter population. All the results suggest that type zSld5-negative type A<small>cl</small> spermatogonia <br />are most undifferentiated type A spermatogonia. <br />  It is generally known that spermatogonial stem cells have ability to restore <br />spermatogenesis when a testis was damaged by a drug or irradiation. To elucidate the<br />characterization of type A spermatogonia, I examined the effect of cytotoxic reagent <br />busulfan on male germ cells. As a result, only type A<small>cl</small> spermatogonia survived after <br />treatment of busulfan, and then recovered spermatogenesis. The morphological <br />observation suggests that type A<small>cl</small> spermatogonia are the most resistant cell population <br />against busulfan in the zebrafish testis. Detailed analysis revealed that the number of <br />zSld5-positive type type A<small>cl</small> spermatogonia increased transiently after busulfan treatment, while the number of type zSld5-negative type A<small>cl</small> spermatogonia relatively stabilized. These results suggest that both zSld5-negative and zSld5-positive type A<small>cl</small> spermatogonia would posses a feature of spermatogonial stem cells, like a actual stem cell and a potential stem cell, respectively. <br />  Furthermore, I developed the <i>in vitro</i> culture condition for supporting the maintenance and proliferation of type A spermatogonia by the use of Sertoli feeder ZtA6-6 cells for 1 month or so. <br />  This study represents the first step towards further understanding the character of spermatogonial stem cells and molecular mechanisms to maintain the undifferentaition status in teleosts.
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