Abstract：
Recently, we reported that, in the naked mole-rat (Heterocephalus glaber) ovary, there is mitotic expansion of the primordial germ cells (PGCs), and the initiation of the meiotic program occurs postnatally. This is opposite to almost all other mammals, including humans and mice, whose reproductive cycle begins very early in development. In both mouse and human, the ovaries become populated with PGCs in utero; these PGCs will later generate the oogonia. After mitotic proliferation, these cells will trigger the meiotic program and initiate meiotic prophase I. Given that all these processes happen in utero, their analysis has been very challenging; so the ability to study them postnatally and to manipulate them with inhibitors or other substances, in the naked mole-rat, opens new possibilities in the field. In this chapter, we present a comprehensive collection of protocols that permit the culture of whole naked mole-rat ovaries, followed by analysis of germ cells, from PGCs to oocytes, in meiotic prophase I, as well the obtention of single-cell suspension or single-nuclei suspension for RNASeq.
摘要：
最近,我们报告说，在裸痣大鼠（异头鼠）卵巢中，原始生殖细胞（PGCs）有丝分裂扩增，减数分裂程序的开始发生在出生后。这与几乎所有其他哺乳动物相反，包括人类和老鼠，其生殖周期在发育早期开始。在老鼠和人类中，卵巢在子宫内充满PGCs;这些PGCs稍后将产生卵原。有丝分裂增殖后，这些细胞将触发减数分裂程序并启动减数分裂前期I。鉴于所有这些过程都发生在子宫内，他们的分析非常具有挑战性；因此，在出生后研究它们并用抑制剂或其他物质操纵它们的能力，在裸露的鼹鼠身上，在该领域开辟了新的可能性。在这一章中,我们提出了一个全面的方案，允许整个裸痣大鼠卵巢的培养，然后分析生殖细胞，从PGCs到卵母细胞，在减数分裂前期I,以及RNASeq的单细胞悬浮液或单核悬浮液的获得。
