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Expression and regulation of regulator of G-protein signaling protein-2 (RGS2) in equine and bovine follicles prior to ovulation: molecular characterization of RGS2 transactivation in bovine granulosa cells.
Abstract: The luteinizing hormone preovulatory surge stimulates several signal pathways essential for ovulation, and the regulator of G-protein signaling protein-2 (RGS2) is thought to be involved in this process. The objectives of this study were to characterize the regulation of RGS2 transcripts in equine and bovine follicles prior to ovulation and to determine its transcriptional control in bovine granulosa cells. To assess the regulation of equine RGS2 prior to ovulation, RT-PCR was performed using total RNA extracted from equine follicles collected at various times after human chorionic gonadotropin (hCG) injection. Results showed that RGS2 mRNA levels were very low at 0 h but markedly increased 12-39 h post-hCG (P < 0.05). In the bovine species, results revealed that RGS2 mRNA levels were low in small and dominant follicles and in ovulatory follicles obtained at 0 h, but markedly increased in ovulatory follicles 6-24 h post-hCG (P < 0.05). To study the molecular control of RGS2 expression, primary cultures of bovine granulosa cells were used. Stimulation with forskolin induced an up-regulation of RGS2 mRNA in vitro. Studies using 5'-deletion mutants identified a minimal region containing full-length basal and forskolin-inducible RGS2 promoter activities. Site-directed mutagenesis indicated that these activities were dependent on CRE and ETS1 cis-elements. Electrophoretic mobility shift assays confirmed the involvement of these elements and revealed their interactions with CREB1 and ETS1 proteins. Chromatin immunoprecipitation assays confirmed endogenous interactions of these proteins with the RGS2 promoter in granulosa cells. Forskolin-inducible RGS2 promoter activity and mRNA expression were markedly decreased by PKA and ERK1/2 inhibitors, and treatment with an antagonist of PGR (RU486) and inhibitors of PTGS2 (NS398) and EGFR (PD153035) blocked the forskolin-dependent RGS2 transcript expression, suggesting the importance of RGS2 in ovulation. Collectively, this study reports for the first time the gonadotropin-dependent up-regulation of RGS2 in equine and bovine preovulatory follicles and presents some of the regulatory controls involved in RGS2 gene expression in granulosa cells.
© 2014 by the Society for the Study of Reproduction, Inc.
Publication Date: 2014-10-22 PubMed ID: 25339105DOI: 10.1095/biolreprod.114.121186Google 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
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This research explores how the regulator of G-protein signaling protein-2 (RGS2) is involved in ovulation in horses and cows, focusing particularly on how RGS2 is regulated in follicles before ovulation and how it is transcriptionally controlled in bovine granulosa cells.
Regulation of RGS2 in Equine and Bovine Follicles
- The study first looked at how RGS2 is regulated in equine follicles prior to ovulation. They used a method called RT-PCR, which allows them to detect specific RNA molecules in the sample, to determine the levels of RGS2.
- They found that RGS2 levels were very low right after the injection of a hormone called hCG, but increased significantly 12-39 hours post-injection. This suggests that the regulation of RGS2 happens within this time frame and is likely related to the ovulation process.
- They observed a similar pattern in cows, with RGS2 levels being low in small and dominant follicles and in ovulatory follicles right after the hormone injection, but increasing significantly 6-24 hours post-injection. This further indicating that the regulation of RGS2 is important in the lead up to ovulation.
Transcriptional Control of RGS2 in Bovine Granulosa Cells
- The researchers then sought to understand the molecular control of RGS2 expression. They used primary cultures of bovine granulosa cells for this purpose.
- They found that stimulating these cells with forskolin, a compound known to activate an enzyme that increases intracellular levels of a molecule called cAMP, resulted in an up-regulation of RGS2. This suggests that forskolin and possibly cAMP levels influence the expression of RGS2.
- Further testing revealed that specific genetic elements, known as CRE and ETS1, are needed for the activities of the RGS2 promoter, which controls the expression of RGS2. Their involvement was confirmed using two different techniques, site-directed mutagenesis and electrophoretic mobility shift assays.
- Finally, they found that certain inhibitors can decrease the forskolin-induced RGS2 promoter activity and mRNA expression, which further supports the idea that the expression and activity of RGS2 play a significant role in the ovulation process.
In conclusion, this research suggests that RGS2 can be up-regulated by hormonal surges during ovulation, and that its transcriptional control in granulosa cells involves specific genetic elements and pathways. This could help in better understanding the mechanisms of ovulation in livestock, which may have implications for improving fertility and reproductive management.
Cite This Article
APA
Sayasith K, Sirois J, Lussier JG.
(2014).
Expression and regulation of regulator of G-protein signaling protein-2 (RGS2) in equine and bovine follicles prior to ovulation: molecular characterization of RGS2 transactivation in bovine granulosa cells.
Biol Reprod, 91(6), 139.
https://doi.org/10.1095/biolreprod.114.121186 Publication
Researcher Affiliations
- Centre de recherche en reproduction animale and the Département de biomédecine vétérinaire, Faculté de médecine vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada k.sayasith@umontreal.ca.
- Centre de recherche en reproduction animale and the Département de biomédecine vétérinaire, Faculté de médecine vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada.
- Centre de recherche en reproduction animale and the Département de biomédecine vétérinaire, Faculté de médecine vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada.
MeSH Terms
- Animals
- Cattle / genetics
- Cattle / physiology
- Cells, Cultured
- Female
- Follicular Phase / drug effects
- Follicular Phase / genetics
- Follicular Phase / metabolism
- Gene Expression Regulation / drug effects
- Gonadotropins / pharmacology
- Granulosa Cells / drug effects
- Granulosa Cells / metabolism
- Horses / genetics
- Horses / physiology
- Ovarian Follicle / drug effects
- Ovarian Follicle / metabolism
- Ovulation / drug effects
- Ovulation / genetics
- Ovulation / metabolism
- RGS Proteins / genetics
- RGS Proteins / metabolism
- Transcriptional Activation / drug effects
Citations
This article has been cited 12 times.- Xu Q, Yao M, Tang C. RGS2 and female common diseases: a guard of women's health. J Transl Med 2023 Aug 30;21(1):583.
- Brady K, Liu HC, Hicks J, Long JA, Porter TE. Global gene expression analysis of the turkey hen hypothalamo-pituitary-gonadal axis during the preovulatory hormonal surge. Poult Sci 2023 Apr;102(4):102547.
- Choi Y, Jeon H, Akin JW, Curry TE, Jo M. The FOS/AP-1 Regulates Metabolic Changes and Cholesterol Synthesis in Human Periovulatory Granulosa Cells. Endocrinology 2021 Sep 1;162(9).
- Selvaraj S, Mondragon-Gonzalez R, Xu B, Magli A, Kim H, Lainé J, Kiley J, Mckee H, Rinaldi F, Aho J, Tabti N, Shen W, Perlingeiro RC. Screening identifies small molecules that enhance the maturation of human pluripotent stem cell-derived myotubes. Elife 2019 Nov 11;8.
- Baufeld A, Koczan D, Vanselow J. L-lactate induces specific genome wide alterations of gene expression in cultured bovine granulosa cells. BMC Genomics 2019 Apr 5;20(1):273.
- Schuermann Y, Rovani MT, Gasperin B, Ferreira R, Ferst J, Madogwe E, Gonçalves PB, Bordignon V, Duggavathi R. ERK1/2-dependent gene expression in the bovine ovulating follicle. Sci Rep 2018 Nov 1;8(1):16170.
- Perschbacher KJ, Deng G, Fisher RA, Gibson-Corley KN, Santillan MK, Grobe JL. Regulators of G protein signaling in cardiovascular function during pregnancy. Physiol Genomics 2018 Aug 1;50(8):590-604.
- Xu SS, Gao L, Xie XL, Ren YL, Shen ZQ, Wang F, Shen M, Eyϸórsdóttir E, Hallsson JH, Kiseleva T, Kantanen J, Li MH. Genome-Wide Association Analyses Highlight the Potential for Different Genetic Mechanisms for Litter Size Among Sheep Breeds. Front Genet 2018;9:118.
- Lussier JG, Diouf MN, Lévesque V, Sirois J, Ndiaye K. Gene expression profiling of upregulated mRNAs in granulosa cells of bovine ovulatory follicles following stimulation with hCG. Reprod Biol Endocrinol 2017 Nov 3;15(1):88.
- Liu DT, Brewer MS, Chen S, Hong W, Zhu Y. Transcriptomic signatures for ovulation in vertebrates. Gen Comp Endocrinol 2017 Jun 1;247:74-86.
- Donadeu FX, Mohammed BT, Ioannidis J. A miRNA target network putatively involved in follicular atresia. Domest Anim Endocrinol 2017 Jan;58:76-83.
- Zaniker EJ, Babayev E, Duncan FE. Common mechanisms of physiological and pathological rupture events in biology: novel insights into mammalian ovulation and beyond. Biol Rev Camb Philos Soc 2023 Oct;98(5):1648-1667.
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