Culture of the Intact Postnatal Naked Mole-Rat Ovary: From Meiotic Prophase to Single-Cell RNASeq

Area of Science:

• Reproductive biology
• Developmental biology
• Comparative genomics

Background:

• Mammalian reproductive development typically initiates in utero, with primordial germ cells (PGCs) populating the fetal ovary.
• These PGCs proliferate mitotically to form oogonia, then enter meiosis I before birth.
• The naked mole-rat (Heterocephalus glaber) presents a unique model where germ cell development occurs postnatally.

Purpose of the Study:

• To establish protocols for culturing whole naked mole-rat ovaries.
• To enable detailed analysis of germ cell development from PGCs to oocytes in meiotic prophase I.
• To facilitate single-cell and single-nuclei suspension preparation for downstream analyses like RNASeq.

Main Methods:

• Whole ovary culture of naked mole-rats.
• Germ cell analysis through various microscopy and molecular techniques.
• Preparation of single-cell and single-nuclei suspensions.

Main Results:

• Successful culture of naked mole-rat ovaries allowing for postnatal study of germ cell development.
• Detailed characterization of germ cells, including PGCs and oocytes in meiotic prophase I.
• Generation of single-cell and single-nuclei suspensions suitable for transcriptomic analysis.

Conclusions:

• The naked mole-rat offers a tractable model for studying mammalian germ cell development due to its unique postnatal timing.
• The provided protocols facilitate in-depth investigation of germ cell biology and reproductive processes in this species.
• This research opens new avenues for understanding conserved and divergent mechanisms in mammalian reproduction.