Rebourcet Diane, Darbey Annalucia, Curley Michael, O'Shaughnessy Peter, Smith Lee B
MRC Centre for Reproductive Health, The Queen's Medical Research Institute, University of Edinburgh, Edinburgh, UK.
Institute of Biodiversity, Animal Health and Comparative Medicine, University of Glasgow, Glasgow, UK.
Methods Mol Biol. 2018;1748:203-228. doi: 10.1007/978-1-4939-7698-0_15.
Testis development and function is regulated by intricate cell-cell cross talk. Characterization of the mechanisms underpinning this has been derived through a wide variety of approaches including pharmacological manipulation, transgenics, and cell-specific ablation of populations. The removal of all or a proportion of a specific cell type has been achieved through a variety of approaches. In this paper, we detail a combined transgenic and pharmacological approach to ablate the Sertoli or germ cell populations using diphtheria toxin in mice. We describe the key steps in generation, validation, and use of the models and also describe the caveats and cautions necessary. We also provide a detailed description of the methodology applied to characterize testis development and function in models of postnatal Sertoli or germ cell ablation.
睾丸的发育和功能受复杂的细胞间相互作用调控。通过多种方法，包括药理学操作、转基因技术以及特定细胞群体的特异性消融，对其潜在机制进行了表征。通过多种方法已实现去除全部或部分特定细胞类型。在本文中，我们详细介绍了一种联合转基因和药理学方法，利用白喉毒素在小鼠中消融支持细胞或生殖细胞群体。我们描述了模型构建、验证和使用的关键步骤，还阐述了必要的注意事项。我们还详细描述了用于表征出生后支持细胞或生殖细胞消融模型中睾丸发育和功能的方法。
