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CCAAT/enhancer-binding proteins alpha and beta directly regulate ovulation and selectively regulate gene expression in granulosa cells by preovulatory stage- and dose-dependent mechanisms

ABSTRACT: The preovulatory luteinizing hormone surge orchestrates complex cellular and molecular events leading to ovulation. CCAAT/enhancer-binding proteins alpha and beta (C/EBPα/β) are transcription factors acutely induced by the luteinizing hormone surge and crucial for ovulation and granulosa cell luteinization. However, biological processes and their regulatory mechanisms downstream of C/EBPα/β in the preovulatory ovary are not completely understood. To address this knowledge gap, we generated Cebpa/bfl/fl;Pgr-Cre mutants and compared them with Cebpa/bfl/fl;Cyp19a1-Cre mutant female mice. The Cebpa/bfl/fl;Cyp19a1-Cre mutants have undetectable levels of C/EBPα/β throughout preovulatory stages and no ovulation, aligning with previous reports, while Cebpa/bfl/fl;Pgr-Cre mutants presented gradual deletion of C/EBPα/β during late preovulatory stage and reduced ovulation rate. Comparison of these two models indicates that sustained expression of C/EBPα/β throughout preovulatory stages is necessary for successful ovulation. This provides a unique opportunity to interrogate gene regulatory mechanisms by C/EBPα/β during different preovulatory time windows and the impact of dysregulating C/EBPα/β on ovulation-associated biological processes. Our study revealed that C/EBPα/β regulate gene expression and distinct biological functions via preovulatory stage-, dose-, and/or potentially isoform ratio- dependent mechanisms. These findings shed new light on the intricate gene regulation mechanisms by C/EBPα/β downstream of the LH surge.

ORGANISM(S): Mus musculus

PROVIDER: GSE279171 | GEO | 2025/05/05

REPOSITORIES: GEO

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Project description:Gonadotropin surge acts on the preovulatory follicle of the ovary to induce luteinization of follicular cells, oocyte meiotic maturation, cumulus expansion and follicular rupture leading to ovulation. These processes are brought about by spatial and temporal changes in transcriptional regulation of genes in the follicular cells in response to the gonadotropin surge. Analysis of gene expression changes in the periovulatory follicular cells will help in delineating the signal transduction pathways involved in the above mentioned processes. In monoovulatory species like bovines, the time interval of 24-28 hours between gonadotropin surge and ovulation provides distinct advantage for studying the temporal changes in the gene expression pattern. Thus, in the present study, we attempt to identify the temporal changes in the global gene expression profile in the periovulatory follicle of buffalo cows in response to gonadotropin surge and the results suggest the involvement of Insulin-like Growth Factor 1 and cytokine signaling pathways in the periovulatory events. Experiment Overall Design: To study the periovulatory gene expression changes in buffalo cows, an induced-ovulation model system involving sequential treatment with PGF2alpha and GnRH was standardized. The follicular wave containing at least one large follicle of ~7mm size was determined by ultrasonography on day 7 of the estrous cycle before administering exogenous PGF2alpha to induce luteolysis and follicular growth. Exogenous GnRH (100Âµg i.m) was administered 36h post PGF2alpha to induce LH surge. The time course of increase in LH levels post GnRH injection was monitored. Since peak LH levels are attained 2 h post GnRH administration, the time intervals of 3 h post GnRH (corresponding to1 h post LH surge) and 24 h post GnRH (corresponding to 22 h post LH surge) were chosen to identify the gene expression profile associated with immediate early and delayed changes in periovulatory follicle respectively. Thus ovaries were collected before, 1 h and 22 h post LH surge and follicle wall and granulosa cells were isolated from the ovaries and snap frozen for the purpose of RNA isolation.

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CCAAT/enhancer-binding proteins alpha and beta directly regulate ovulation and selectively regulate gene expression in granulosa cells by preovulatory stage- and dose-dependent mechanisms

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