Department of Anatomy and Cell Biology, Brody School of Medicine, East Carolina University, Greenville, NC 27834, USA.
Flow Cytometry Facility, Brody School of Medicine, East Carolina University, Greenville, NC 27834, USA.
Development. 2022 Nov 15;149(22). doi: 10.1242/dev.200713. Epub 2022 Nov 16.
In mammalian testes, premeiotic spermatogonia respond to retinoic acid by completing an essential lengthy differentiation program before initiating meiosis. The molecular and cellular changes directing these developmental processes remain largely undefined. This wide gap in knowledge is due to two unresolved technical challenges: (1) lack of robust and reliable in vitro models to study differentiation and meiotic initiation; and (2) lack of methods to isolate large and pure populations of male germ cells at each stage of differentiation and at meiotic initiation. Here, we report a facile in vitro differentiation and meiotic initiation system that can be readily manipulated, including the use of chemical agents that cannot be safely administered to live animals. In addition, we present a transgenic mouse model enabling fluorescence-activated cell sorting-based isolation of millions of spermatogonia at specific developmental stages as well as meiotic spermatocytes.
在哺乳动物的睾丸中，精原细胞在启动减数分裂之前，通过完成一个必要的、漫长的分化程序来响应视黄酸。指导这些发育过程的分子和细胞变化在很大程度上仍未得到明确界定。造成这一知识鸿沟的原因有两个尚未解决的技术挑战：（1）缺乏用于研究分化和减数分裂起始的稳健可靠的体外模型；（2）缺乏在分化和减数分裂起始的每个阶段分离大量和纯的雄性生殖细胞群体的方法。在这里，我们报告了一种简单的体外分化和减数分裂起始系统，该系统易于操作，包括使用无法安全施用于活体动物的化学试剂。此外，我们还展示了一种转基因小鼠模型，该模型能够基于荧光激活细胞分选分离特定发育阶段以及减数分裂精母细胞的数百万个精原细胞。
