Expression and regulation of transcripts encoding two members of the NR5A nuclear receptor subfamily of orphan nuclear receptors, steroidogenic factor-1 and NR5A2, in equine ovarian cells during the ovulatory process.
Abstract: Steroidogenic factor-1 (SF-1, NR5A1a) is a member of the NR5A nuclear receptor subfamily and has been implicated as a key transcriptional regulator of all ovarian steroidogenic genes in vitro. To establish links between the expression of SF-1 and that of the steroidogenic genes in vivo, the objectives of this study were to clone equine SF-1 and examine the regulation of its messenger RNA (mRNA) in follicular cells during human CG (hCG)-induced ovulation. The equine SF-1 primary transcript was cloned by a combination of RT-PCR techniques. Results showed that the transcript was composed of a 5'-untranslated region (UTR) of 161 bp, an open reading frame (ORF) of 1386 bp that encodes a highly-conserved 461-amino acid protein, and a 3'-UTR of 518 bp. The cloning of SF-1 also led to the unexpected and serendipitous isolation of the highly-related orphan nuclear receptor NR5A2, which was shown to include a 5'-UTR of 243 bp, an ORF of 1488 bp, and a 3'-UTR of 1358 bp. The NR5A2 ORF encodes a 495-amino acid protein that is 60% identical to SF-1, including 99%-similar DNA-binding domains. Northern blot analysis revealed that SF-1 and NR5A2 were expressed in all major steroidogenic tissues, with the exception that NR5A2 was not present in the adrenal. Interestingly, NR5A2 was found to be, by far, the major NR5A subfamily member expressed in the preovulatory follicle and the corpus luteum. Using a semiquantitative RT-PCR/Southern blotting approach, the regulation of SF-1 and NR5A2 mRNAs in vivo was studied in equine follicular cells obtained from preovulatory follicles isolated between 0 and 39 h post hCG. Results showed that the theca interna was the predominant site of SF-1 mRNA expression in the follicle, and that hCG caused a significant decrease in SF-1 levels between 12-39 h in theca interna and between 24-39 h post hCG in granulosa cells (P < 0.05). In contrast, the granulosa cell layer was the predominant, if not the sole, site of NR5A2 mRNA expression in the follicle. Importantly, NR5A2 was much more highly expressed in granulosa cells than SF-1. The administration of hCG caused a significant decrease in NR5A2 transcripts in granulosa cells at 30, 36, and 39 h post hCG (P < 0.05). Thus, this study is the first to report the concomitant regulation of SF-1 in theca interna and granulosa cells throughout the ovulation/luteinization process, and to demonstrate the novel expression and hormonal regulation of NR5A2 in ovarian cells. Based on the marked expression of NR5A2 in equine granulosa and luteal cells and on mounting evidence of a functional redundancy between SF-1 and NR5A2 in other species, it is proposed that NR5A2 may play a key role in the regulation of gonadal steroidogenic gene expression.
Publication Date: 2000-12-07 PubMed ID: 11108279DOI: 10.1210/endo.141.12.7808Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This research investigates the role of two members of the NR5A nuclear receptor subfamily, steroidogenic factor-1 (SF-1) and NR5A2, specifically looking at how they affect gene expression in horse’s ovarian cells during ovulation. The study shows how human chorionic gonadotropin hormone (hCG) influences these factors and hints at the possibility of NR5A2 playing a crucial role in the regulation of steroidogenic genes.
Methods and Findings
- The study began with the cloning of the equine SF-1 and observing the regulation of its messenger RNA (mRNA) in follicular cells during human CG (hCG)-induced ovulation. This cloning process unexpectedly also led to the isolation of the highly-related orphan nuclear receptor NR5A2.
- Both SF-1 and NR5A2 were inspected and found to be expressed in all major steroidogenic tissues, except for NR5A2, which wasn’t present in the adrenal. Remarkably, NR5A2 was predominantly present in the preovulatory follicle and the corpus luteum, more so than SF-1.
- The study then moved on to examine the expressions of SF-1 and NR5A2 mRNAs in equine follicular cells obtained from preovulatory follicles. The expression of SF-1 mRNA was mainly found in the theca interna of the follicle and significantly decreased after hCG were given. In contrast, the granulosa cell layer was the primary site of NR5A2 mRNA expression in the follicle. Moreover, NR5A2 dramatically showed a much higher expression than SF-1.
Key Conclusion
- The study confirmed the simultaneous regulation of SF-1 in the theca interna and granulosa cells throughout the ovulation/luteinization process and highlighted the new discovery of the expression and hormonal regulation of NR5A2 in ovarian cells.
- Given the high expression of NR5A2 in equine granulosa and luteal cells and the evidence of functional redundancy between SF-1 and NR5A2 from other species, the study suggested that NR5A2 might play a significant role in regulating gonadal steroidogenic gene expression.
Cite This Article
APA
Boerboom D, Pilon N, Behdjani R, Silversides DW, Sirois J.
(2000).
Expression and regulation of transcripts encoding two members of the NR5A nuclear receptor subfamily of orphan nuclear receptors, steroidogenic factor-1 and NR5A2, in equine ovarian cells during the ovulatory process.
Endocrinology, 141(12), 4647-4656.
https://doi.org/10.1210/endo.141.12.7808 Publication
Researcher Affiliations
- Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada.
MeSH Terms
- Amino Acid Sequence
- Animals
- Base Sequence
- Chorionic Gonadotropin / pharmacology
- Cloning, Molecular
- DNA, Complementary / chemistry
- DNA, Complementary / genetics
- DNA-Binding Proteins / chemistry
- DNA-Binding Proteins / genetics
- Female
- Fushi Tarazu Transcription Factors
- Gene Expression Regulation
- Granulosa Cells / metabolism
- Homeodomain Proteins
- Horses / metabolism
- Molecular Sequence Data
- Ovary / metabolism
- Ovulation
- Proteins / analysis
- Proteins / chemistry
- Proteins / genetics
- RNA, Messenger / analysis
- Receptors, Cytoplasmic and Nuclear / genetics
- Steroidogenic Factor 1
- Theca Cells / metabolism
- Transcription Factors / chemistry
- Transcription Factors / genetics
Citations
This article has been cited 25 times.- Sun W, Shi Q, Li J, Li J, Yu L. LRH1 Promotes Tumor Cell Proliferation and Migration and Is Correlated With Poor Prognosis in Ovarian Cancer.. Front Oncol 2020;10:583566.
- Kim Y, Ghil S. Regulators of G-protein signaling, RGS2 and RGS4, inhibit protease-activated receptor 4-mediated signaling by forming a complex with the receptor and Gα in live cells.. Cell Commun Signal 2020 Jun 9;18(1):86.
- Zhu G, Fang C, Li J, Mo C, Wang Y, Li J. Transcriptomic Diversification of Granulosa Cells during Follicular Development in Chicken.. Sci Rep 2019 Apr 2;9(1):5462.
- Wu C, Feng J, Li L, Wu Y, Xie H, Yin Y, Ye J, Li Z. Liver receptor homologue 1, a novel prognostic marker in colon cancer patients.. Oncol Lett 2018 Sep;16(3):2833-2838.
- Aaronson Y, Livyatan I, Gokhman D, Meshorer E. Systematic identification of gene family regulators in mouse and human embryonic stem cells.. Nucleic Acids Res 2016 May 19;44(9):4080-9.
- Kramer HB, Lai CF, Patel H, Periyasamy M, Lin ML, Feller SM, Fuller-Pace FV, Meek DW, Ali S, Buluwela L. LRH-1 drives colon cancer cell growth by repressing the expression of the CDKN1A gene in a p53-dependent manner.. Nucleic Acids Res 2016 Jan 29;44(2):582-94.
- Montanaro D, Santoro M, Carpino A, Perrotta I, De Amicis F, Sirianni R, Rago V, Gervasi S, Aquila S. Human sperm liver receptor homolog-1 (LRH-1) acts as a downstream target of the estrogen signaling pathway.. J Anat 2015 Oct;227(4):541-9.
- Nadolny C, Dong X. Liver receptor homolog-1 (LRH-1): a potential therapeutic target for cancer.. Cancer Biol Ther 2015;16(7):997-1004.
- Zhao Y, Liu W, Hua M, Shi R, Wang H, Yang W. Relationship between CATSPERB, NR5A2 gene polymorphisms and Peak Bone Mineral Density in College Students in China.. Iran J Public Health 2014 Aug;43(8):1060-9.
- Udhane S, Kempna P, Hofer G, Mullis PE, Flück CE. Differential regulation of human 3β-hydroxysteroid dehydrogenase type 2 for steroid hormone biosynthesis by starvation and cyclic AMP stimulation: studies in the human adrenal NCI-H295R cell model.. PLoS One 2013;8(7):e68691.
- Wang Q, Leader A, Tsang BK. Follicular stage-dependent regulation of apoptosis and steroidogenesis by prohibitin in rat granulosa cells.. J Ovarian Res 2013 Apr 8;6(1):23.
- Mellado-Gil JM, Cobo-Vuilleumier N, Gauthier BR. Islet β-Cell Mass Preservation and Regeneration in Diabetes Mellitus: Four Factors with Potential Therapeutic Interest.. J Transplant 2012;2012:230870.
- Matulis CK, Mayo KE. The LIM domain protein FHL2 interacts with the NR5A family of nuclear receptors and CREB to activate the inhibin-α subunit gene in ovarian granulosa cells.. Mol Endocrinol 2012 Aug;26(8):1278-90.
- Yumoto F, Nguyen P, Sablin EP, Baxter JD, Webb P, Fletterick RJ. Structural basis of coactivation of liver receptor homolog-1 by β-catenin.. Proc Natl Acad Sci U S A 2012 Jan 3;109(1):143-8.
- Fernandez-Marcos PJ, Auwerx J, Schoonjans K. Emerging actions of the nuclear receptor LRH-1 in the gut.. Biochim Biophys Acta 2011 Aug;1812(8):947-55.
- Yang FM, Lin YC, Hu MC. Identification of two functional nuclear localization signals mediating nuclear import of liver receptor homologue-1.. Cell Mol Life Sci 2011 Apr;68(7):1241-53.
- Zhang YP, Deng FY, Chen Y, Pei YF, Fang Y, Guo YF, Guo X, Liu XG, Zhou Q, Liu YJ, Deng HW. Replication study of candidate genes/loci associated with osteoporosis based on genome-wide screening.. Osteoporos Int 2010 May;21(5):785-95.
- Andreu-Vieyra C, Chen R, Matzuk MM. Conditional deletion of the retinoblastoma (Rb) gene in ovarian granulosa cells leads to premature ovarian failure.. Mol Endocrinol 2008 Sep;22(9):2141-61.
- Mataki C, Magnier BC, Houten SM, Annicotte JS, Argmann C, Thomas C, Overmars H, Kulik W, Metzger D, Auwerx J, Schoonjans K. Compromised intestinal lipid absorption in mice with a liver-specific deficiency of liver receptor homolog 1.. Mol Cell Biol 2007 Dec;27(23):8330-9.
- Mendelson CR, Kamat A. Mechanisms in the regulation of aromatase in developing ovary and placenta.. J Steroid Biochem Mol Biol 2007 Aug-Sep;106(1-5):62-70.
- Li Y, Choi M, Suino K, Kovach A, Daugherty J, Kliewer SA, Xu HE. Structural and biochemical basis for selective repression of the orphan nuclear receptor liver receptor homolog 1 by small heterodimer partner.. Proc Natl Acad Sci U S A 2005 Jul 5;102(27):9505-10.
- Wang W, Zhang C, Marimuthu A, Krupka HI, Tabrizizad M, Shelloe R, Mehra U, Eng K, Nguyen H, Settachatgul C, Powell B, Milburn MV, West BL. The crystal structures of human steroidogenic factor-1 and liver receptor homologue-1.. Proc Natl Acad Sci U S A 2005 May 24;102(21):7505-10.
- Schoonjans K, Dubuquoy L, Mebis J, Fayard E, Wendling O, Haby C, Geboes K, Auwerx J. Liver receptor homolog 1 contributes to intestinal tumor formation through effects on cell cycle and inflammation.. Proc Natl Acad Sci U S A 2005 Feb 8;102(6):2058-62.
- Dell'Aquila ME, Caillaud M, Maritato F, Martoriati A, Gérard N, Aiudi G, Minoia P, Goudet G. Cumulus expansion, nuclear maturation and connexin 43, cyclooxygenase-2 and FSH receptor mRNA expression in equine cumulus-oocyte complexes cultured in vitro in the presence of FSH and precursors for hyaluronic acid synthesis.. Reprod Biol Endocrinol 2004 Jun 22;2:44.
- Suzuki T, Kasahara M, Yoshioka H, Morohashi K, Umesono K. LXXLL-related motifs in Dax-1 have target specificity for the orphan nuclear receptors Ad4BP/SF-1 and LRH-1.. Mol Cell Biol 2003 Jan;23(1):238-49.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
