Identification of FSH-regulated and estrous stage–specific transcriptional networks in mouse ovaries
Edited by Thomas Spencer, University of Missouri, Columbia, MO; received June 14, 2024; accepted January 3, 2025
Significance
Despite decades of work on follicle-stimulating hormone (FSH) action in ovaries, how FSH regulates transcriptional networks in vivo has not been studied. Here, using RNA-Seq analysis on ovaries obtained from mice with intact FSH, no FSH, or FSH rescue, we have identified FSH-responsive genes that are estrous specifically regulated in vivo. Our studies may eventually allow us to identify potential targets for fertility enhancement or blockade in women.
Abstract
Follicle-stimulating hormone (FSH) acts by binding to FSHRs expressed on ovarian granulosa cells and produces estradiol. FSH is essential for female fertility because mice lacking FSH (Fshb KO) are anestrous and infertile. Although several in vitro cell culture and ex vivo approaches combined with pharmacological hormone treatment were used to identify FSH-regulated genes, how FSH orchestrates ovarian gene networks in vivo has not been investigated. Whether FSH-regulated genes display estrous stage–specific expression changes has also not been studied. Here, we functionally rescued Fshb null mice with a gonadotrope-targeted HFSHB transgene and performed RNA-Seq analysis on ovarian RNAs obtained from FSH-intact (WT), FSH-deficient (Fshb KO), and FSH-rescue (HFSHB+ rescue) mice. By comparing WT vs. Fshb KO and Fshb KO vs. HFSHB+ rescue ovarian gene expression datasets, we identified FSH-responsive genes in vivo. Cross interrogation of these datasets further allowed us to identify several transcription factors (TFs) and RNA-binding proteins specific to FSH-regulated genes. In an independent set of experiments, we performed RNA-Seq analysis on ovarian RNAs from mice in diestrous (DE), proestrous (PE), and estrous (E) and identified estrous stage–specific ovarian gene expression patterns. Interestingly, many of the FSH-regulated TFs themselves were estrous-stage specifically expressed. We found that ESR2 and GATA6, two known FSH-responsive TFs, and their target genes are reciprocally regulated with distinct patterns of expression in estrous stages. Together, our in vivo models and RNA-Seq analyses identify FSH-regulated ovarian genes in specific estrous stages that are under transcriptional and posttranscriptional control.
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Data, Materials, and Software Availability
RNA-Seq data have been uploaded to the Gene Expression Omnibus database (Accession No. GSM8288804) (47). All other data are included in the manuscript and/or supporting information.
Acknowledgments
This work was supported in part by the University of Colorado Anschutz Medical Campus RNA Bioscience Initiative (N.M.), Boettcher Foundation Webb-Waring Early Career Investigator Award (N.M.), and NIH Grants GM147025 (N.M.) and HD103384, AG029531 (T.R.K.), The Makowski Family Endowment (T.R.K.), Gonadotropin Research Fund (T.R.K.), and Global Consortium on Reproductive Longevity and Equity pilot Grant-1323 (T.R.K.). K.W. is a recipient of the NSF Graduate Research Fellowship (Grant No. 1000317291).
Author contributions
N.M. and T.R.K. designed research; K.W., A.B., Z.L., M.L., and T.R.K. performed research; K.W., Z.L., N.M., and T.R.K. analyzed data; N.M. and T.R.K. obtained funding; and N.M. and T.R.K. wrote the paper.
Competing interests
The authors declare no competing interest.
Supporting Information
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Copyright © 2025 the Author(s). Published by PNAS. This article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND).
Data, Materials, and Software Availability
RNA-Seq data have been uploaded to the Gene Expression Omnibus database (Accession No. GSM8288804) (47). All other data are included in the manuscript and/or supporting information.
Submission history
Received: June 14, 2024
Accepted: January 3, 2025
Published online: February 10, 2025
Published in issue: February 18, 2025
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Acknowledgments
This work was supported in part by the University of Colorado Anschutz Medical Campus RNA Bioscience Initiative (N.M.), Boettcher Foundation Webb-Waring Early Career Investigator Award (N.M.), and NIH Grants GM147025 (N.M.) and HD103384, AG029531 (T.R.K.), The Makowski Family Endowment (T.R.K.), Gonadotropin Research Fund (T.R.K.), and Global Consortium on Reproductive Longevity and Equity pilot Grant-1323 (T.R.K.). K.W. is a recipient of the NSF Graduate Research Fellowship (Grant No. 1000317291).
Author contributions
N.M. and T.R.K. designed research; K.W., A.B., Z.L., M.L., and T.R.K. performed research; K.W., Z.L., N.M., and T.R.K. analyzed data; N.M. and T.R.K. obtained funding; and N.M. and T.R.K. wrote the paper.
Competing interests
The authors declare no competing interest.
Notes
This article is a PNAS Direct Submission.
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Identification of FSH-regulated and estrous stage–specific transcriptional networks in mouse ovaries, Proc. Natl. Acad. Sci. U.S.A.
122 (7) e2411977122,
https://doi.org/10.1073/pnas.2411977122
(2025).
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