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: 2007
• 资助国家: 加拿大
• 起止时间: 2007-01-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=364326
• 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（spermatogonialstemcells，Sg）是精子的成体前体细胞;精原细胞移植是一种从供体睾丸中分离出Sg干细胞并将其注射到受体的生精小管中以由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有价值的新工具，并且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而运输遗传学的额外方法，以及将基因直接引入生殖系的成本较低的方法。公羊是研究哺乳动物精原细胞出生后发育的一种很好的替代啮齿动物的动物。公羊相对较小，价格便宜，公羊羔羊不需要阉割，因此，屠宰场有现成的各种年龄的睾丸供研究之用。在研究干细胞Sg时必须克服的一个主要问题是，它们在睾丸中的数量很少。因此，为了寻找重要的分子，他们的健康，我们将用激光从公羊和羔羊睾丸的冰冻切片中收集单个Sg干细胞，然后从细胞中提取RNA，通过PCR对重要基因转录本进行定量。RNA也将用于使用微阵列对这些细胞表达的基因进行广泛的筛选。这些信息将用于提高公羊精原细胞移植的效率。我们还将开发一种方法，最后，我们将利用精原细胞移植和转导的供体干细胞Sg来检测筛选出的基因在公羊精原细胞发育中的功能，在这个过程中，我们将证明精原细胞移植的效率，为农场动物工业和研究人员寻求大型动物疾病模型。