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bioRxiv · 10.1101/2024.02.08.579446

Transcriptomic Signatures of WNT-Driven Pathways and Granulosa Cell-Oocyte Interactions during Primordial Follicle Activation

Takase, H. M.·Mishina, T.·Hayashi, T.·Yoshimura, M.·Kuse, M.·Nikaido, I.·Kitajima, T. S.
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Abstract

Primordial follicle activation (PFA) is a pivotal event in female reproductive biology, coordinating the transition from quiescent to growing follicles. This study employed comprehensive single-cell RNA sequencing to gain insights into the detailed regulatory mechanisms governing the synchronized dormancy and activation between granulosa cells (GCs) and oocytes with the progression of the PFA process. Wntless (Wls) conditional knockout (cKO) mice served as a unique model, suppressing the transition from pre-GCs to GCs, and disrupting somatic cell-derived WNT signaling in the ovary. Our data revealed immediate transcriptomic changes in GCs post-PFA in Wls cKO mice, leading to a divergent trajectory, while oocytes exhibited modest transcriptomic alterations. Subpopulation analysis identified the molecular pathways affected by WNT signaling on GC maturation, along with specific gene signatures linked to dormant and activated oocytes. Despite minimal evidence of continuous up-regulation of dormancy-related genes in oocytes, the loss of WNT signaling in (pre-)GCs impacted gene expression in oocytes even before PFA, subsequently influencing them globally. The infertility observed in Wls cKO mice was attributed to compromised GC-oocyte molecular crosstalk and the microenvironment for oocytes. Our study highlights the pivotal role of the WNT-signaling pathway and its molecular signature, emphasizing the importance of intercellular crosstalk between (pre-)GCs and oocytes in orchestrating folliculogenesis. Author summaryIn mammalian ovaries, primordial follicles protect dormant oocytes from wasteful consumption. We explored the complex molecular dynamics of the critical process of primordial follicle activation (PFA) that underlies female reproductive health, with a focus on the role of WNT signaling. By utilizing a unique mouse model with disrupted WNT signaling, our study reveals novel insights into the regulatory dynamics of granulosa cells (GCs) and oocytes during PFA. Single-cell transcriptome profiling uncovered distinct cell populations and highlighted the impact of WNT signaling on ovarian cell types. This study reveals distinct transcriptomic changes in GCs post-PFA, which affects their differentiation trajectories and highlights the critical role of WNT signaling in establishing GC function. We further examined the consequences of disrupted microenvironments on oocytes and GC-oocyte interactions, which elucidated altered oocyte subclusters and potential links to infertility. These findings provide a foundation for understanding the connections between WNT signaling and ovarian function. Our comprehensive analysis contributes valuable knowledge to reproductive biology, emphasizing the clinical relevance of WNT signaling in female fertility.

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Takase, H. M., Mishina, T., Hayashi, T., Yoshimura, M., Kuse, M., Nikaido, I., Kitajima, T. S.. 2024-02-11. Transcriptomic Signatures of WNT-Driven Pathways and Granulosa Cell-Oocyte Interactions during Primordial Follicle Activation. https://doi.org/10.1101/2024.02.08.579446

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