The biology of spermatogonial development in sheep

绵羊精原细胞发育的生物学

• 批准号: 138054-2006
• 负责人: Hahnel, Ann
• 金额: $ 1.82万
• 依托单位: University of Guelph
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2008
• 资助国家: 加拿大
• 起止时间: 2008-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=391417
• 关键词: biology; spermatogonial; development; sheep

Spermatogonial stem cells (stem Sg) are the adult precursors of sperm; their health is critical to propagation of species.  Spermatogonial transplantation is a technique that isolates stem Sg from donor testes and injects them into the seminiferous tubules of a recipient for production of donor sperm by the recipient animal.  Spermatogonial transplantation in mice has proven to be a valuable new tool for studying spermatogenesis, and has opened the possibility of using stem Sg for preservation and dissemination of genetics.  In particular the sheep industry would benefit from additional ways to transport genetics without shipping animals, and from less costly ways to introduce genes directly into the germ line. Rams are a good alternative to rodents for studying post-natal development of spermatogonia in mammals.  They are comparatively small and cheap, and ram lambs are not castrated, so there is a ready supply of testes of all ages from the abattoir for study.  A major problem that must be overcome in studying stem Sg is that they are a minor population in the testis.  Therefore in order to look for molecules important to their health, we will collect individual stem Sg from frozen sections of ram lamb testes with a laser and then extract RNA from the cells to quantify important gene transcripts by PCR.  The RNA also will be used for a broad screening of genes expressed by these cells using microarrays.  The information will be used to increase the efficiency of spermatogonial transplantation in rams.  We also will develop a way to permanently incorporate genes into donor stem Sg using a retrovirus.  Finally we will use spermatogonial transplantation and transduced donor stem Sg to test the function of genes identified in the screens in development of ram spermatogonia.  In the process, we will demonstrate efficiency of spermatogonial transplantation for the farm animal industries and for researchers seeking large animal models of disease.

精原干细胞（stem Sg）是精子的成体前体细胞；它们的健康对于物种的繁殖至关重要。  精原细胞移植是一种从供体睾丸中分离出干细胞并将其注射到受体的生精小管中，以便受体动物产生供体精子的技术。  小鼠精原细胞移植已被证明是研究精子发生的一种有价值的新工具，并开启了利用干细胞保存和传播遗传学的可能性。  特别是，绵羊产业将受益于无需运输动物即可运输基因的其他方法，以及将基因直接引入种系的成本较低的方法。公羊是研究哺乳动物产后精原细胞发育的啮齿类动物的良好替代品。  它们相对较小且便宜，而且公羊羔羊没有被阉割，因此屠宰场有现成的各个年龄段的睾丸可供研究。  研究茎 Sg 必须克服的一个主要问题是它们是睾丸中的少数群体。  因此，为了寻找对其健康重要的分子，我们将用激光从公羊睾丸冷冻切片中收集单个茎Sg，然后从细胞中提取RNA，通过PCR定量重要基因转录本。  RNA 还将用于使用微阵列广泛筛选这些细胞表达的基因。  该信息将用于提高公羊精原细胞移植的效率。  我们还将开发一种使用逆转录病毒将基因永久整合到供体茎 Sg 中的方法。  最后，我们将使用精原细胞移植和转导的供体干细胞 Sg 来测试筛选中鉴定的基因在公羊精原细胞发育中的功能。  在此过程中，我们将展示精原细胞移植对于农场动物产业和寻求大型动物疾病模型的研究人员的效率。