MicroRNA regulation of cyclooxygenase-2 during embryo implantation - PubMed (original) (raw)
MicroRNA regulation of cyclooxygenase-2 during embryo implantation
Anindita Chakrabarty et al. Proc Natl Acad Sci U S A. 2007.
Abstract
The implantation process is complex, requiring reciprocal interactions between implantation-competent blastocysts and the receptive uterus. Because microRNAs (miRNAs) have major roles in regulating gene expression, we speculated that they participate in directing the highly regulated spatiotemporally expressed genetic network during implantation. Here, we show that two miRNAs, mmu-miR-101a and mmu-miR-199a*, are spatiotemporally expressed in the mouse uterus during implantation coincident with expression of cyclooxygenase-2, a gene critical for implantation. More interestingly, our in vitro gain- and loss-of-function experiments show that cyclooxygenase-2 expression is posttranscriptionally regulated by these two miRNAs. We report on miRNA-mediated regulation of uterine gene expression in the context of implantation. We believe that many other critical genes related to this process are also regulated by miRNAs. Thus, elucidating the physiological roles of uterine miRNAs will help us better understand the genetic control of implantation, the gateway to a successful pregnancy.
Conflict of interest statement
The authors declare no conflict of interest.
Figures
Fig. 1.
miRNA biogenesis in periimplantation mouse uteri. (A) miRNA profiling with oligonucleotide microarray analysis of day-1 (D1) and day-4 (D4) pregnant uteri. Yellow spots along the diagonal line represent miRNAs with similar expression levels between days 1 and 4. Orange spots indicate up-regulated miRNAs in prereceptive uteri (day 1), and blue spots indicate up-regulated miRNAs in receptive uteri (day 4). (B) RT-PCR of Dicer1 and Ago1–4 in periimplantation uteri. β-Actin serves as a control. (C) Western blotting of Dicer1 protein. Mouse ovary, positive controls; Actin, loading control. IS, implantation site; IIS, interimplantation site.
Fig. 2.
Mature miR-199a* is expressed in periimplantation mouse uteri. (A) Northern hybridization with 32P-labeled probe in total uterine RNA samples on day 1 (D1) and day 4 (D4) of pregnancy. The ethidium bromide-stained gel served as a loading control. (B) In situ hybridization of uterine cross-sections with DIG-labeled LNA probe on days 1, 4, 5, and 6 of pregnancy. Arrows indicate locations of blastocysts. le, luminal epithelium; ge, glandular epithelium; s, stroma; myo, myometrium; M, mesometrial pole; AM, antimesometrial pole; IS, implantation site. (Scale bar: 400 μm.)
Fig. 3.
Mature miR-101a is expressed in periimplantation mouse uteri. (A) Northern hybridization of total uterine RNA samples from day-1 (D1) and day-4 (D4) pregnant uteri. The ethidium bromide-stained gel served as a loading control. (B) In situ hybridization of uterine cross-sections with DIG-labeled LNA probe on days 1, 4, 5, and 6 of pregnancy. Arrows indicate the location of blastocysts. Abbreviations are as in Fig. 2. (Scale bar: 400 μm.)
Fig. 4.
Uterine expression of miR-101a, miR-199a*, and Cox-2. (A) In situ hybridization of Cox-2 with 35S-labeled probe (Left) and protein by immunohistochemistry (Right) in serial cross-sections from day-5 implantation sites (IS). (B) In situ hybridization of miR-199a* with DIG-labeled LNA probe on longitudinal sections from delayed and implanting uteri. Arrows indicate the location of blastocysts. (C) Uterine Cox-2 and miR-101a levels during experimentally induced decidualization. Western blot shows Cox-2 protein levels at time points after intrauterine oil infusion (+); noninfused contralateral uterine horns serve as negative controls (−). Actin is a loading control. The ethidium bromide-stained gels served as loading control for miR-101a Northern blot. (Scale bars: 200 μm, A; 400 μm, B.)
Fig. 5.
Cross-species homology of miRNAs targeting COX-2 mRNA. (A and B) The structure and homology of the predicted interactions between Cox-2 mRNA and miR-101 (A) and miR-199a* (B). (C) The predicted annealing sites of the seed and 3′ terminus of these miRNAs are well conserved in Cox-2 mRNA.
Fig. 6.
miR-199a* and miR-101a regulate Cox-2 translation. (A) Renilla Luc-m_Cox-2_-3′ UTR reporter construct. (B) Concentration-dependent decreases in reporter activity with increasing doses of precursor miR-199a* or miR-101a in HEK293 cells. A scrambled miRNA had no effect on reporter activity (control). (C) Antisense inhibitors of miR-199a* and miR-101a relieve miRNA-mediated repression of _Cox-2_-3′ UTR-Luc reporter activity; scrambled antisense sequence (control) had no effect. Each transfection was performed in triplicate. Data are presented as relative Luc activity (mean ± SEM). (D) miR-199a* and miR-101a repress endogenous Cox-2 translation. Concentration-dependent decreases in endogenous Cox-2 protein levels in HeLa cells 72 h after transfection with miR-199a* or miR-101a precursors. Western blotting of Cox-2 (Upper) and corresponding densitometry analyses (Lower) are shown. Actin, loading control.
Similar articles
- MicroRNA-199a mediates mucin 1 expression in mouse uterus during implantation.
Inyawilert W, Fu TY, Lin CT, Tang PC. Inyawilert W, et al. Reprod Fertil Dev. 2014 Jun;26(5):653-64. doi: 10.1071/RD12097. Reprod Fertil Dev. 2014. PMID: 23759257 - Mmu-microRNA-200a overexpression leads to implantation defect by targeting phosphatase and tensin homolog in mouse uterus.
Shen LJ, He JL, Yang DH, Ding YB, Chen XM, Geng YQ, Liu SJ, Liu XQ, Wang YX. Shen LJ, et al. Reprod Sci. 2013 Dec;20(12):1518-28. doi: 10.1177/1933719113488453. Epub 2013 May 20. Reprod Sci. 2013. PMID: 23690337 Free PMC article. - Mmu-miR-193 is involved in embryo implantation in mouse uterus by regulating GRB7 gene expression.
Li R, He J, Chen X, Ding Y, Wang Y, Long C, Shen L, Liu X. Li R, et al. Reprod Sci. 2014 Jun;21(6):733-42. doi: 10.1177/1933719113512535. Epub 2013 Dec 13. Reprod Sci. 2014. PMID: 24336674 Free PMC article. - The role of microRNAs in human embryo implantation: a review.
Paul ABM, Sadek ST, Mahesan AM. Paul ABM, et al. J Assist Reprod Genet. 2019 Feb;36(2):179-187. doi: 10.1007/s10815-018-1326-y. Epub 2018 Oct 12. J Assist Reprod Genet. 2019. PMID: 30315515 Free PMC article. Review. - MicroRNA and implantation.
Galliano D, Pellicer A. Galliano D, et al. Fertil Steril. 2014 Jun;101(6):1531-44. doi: 10.1016/j.fertnstert.2014.04.023. Fertil Steril. 2014. PMID: 24882617 Review.
Cited by
- The histone methyltransferase KMT2D is essential for embryo implantation via regulating precise differentiation of endometrial cells.
Kobayashi R, Tajika Y, Kohmaru J, Morita S, Horii T, Mizukami Y, Aikawa S, Hirota Y, Hatada I. Kobayashi R, et al. Cell Death Discov. 2024 Aug 8;10(1):357. doi: 10.1038/s41420-024-02134-9. Cell Death Discov. 2024. PMID: 39117610 Free PMC article. - Expression of circulating oar-miR-485-5p and oar-miR-493-5p during the estrous cycle and early pregnancy in ovine plasma.
Ucar EH, Hitit M, Kose M, Atli MO. Ucar EH, et al. Anim Reprod. 2024 Mar 4;21(1):e20230115. doi: 10.1590/1984-3143-AR2023-0115. eCollection 2024. Anim Reprod. 2024. PMID: 38510567 Free PMC article. - TNFα-Induced Altered miRNA Expression Links to NF-κB Signaling Pathway in Endometriosis.
Banerjee S, Xu W, Doctor A, Driss A, Nezhat C, Sidell N, Taylor RN, Thompson WE, Chowdhury I. Banerjee S, et al. Inflammation. 2023 Dec;46(6):2055-2070. doi: 10.1007/s10753-023-01862-x. Epub 2023 Jun 30. Inflammation. 2023. PMID: 37389684 Free PMC article. - TNFα-induced altered miRNA expression links to NF-κB signaling pathway in endometriosis.
Banerjee S, Xu W, Doctor A, Driss A, Nezhat C, Sidell N, Taylor RN, Thompson WE, Chowdhury I. Banerjee S, et al. Res Sq [Preprint]. 2023 May 4:rs.3.rs-2870585. doi: 10.21203/rs.3.rs-2870585/v1. Res Sq. 2023. PMID: 37205467 Free PMC article. Updated. Preprint. - Nonsteroidal anti-inflammatory drugs and implications for the cyclooxygenase pathway in embryonic development.
Leathers TA, Rogers CD. Leathers TA, et al. Am J Physiol Cell Physiol. 2023 Feb 1;324(2):C532-C539. doi: 10.1152/ajpcell.00430.2022. Epub 2023 Jan 9. Am J Physiol Cell Physiol. 2023. PMID: 36622071 Free PMC article. Review.
References
- Nilsen TW. Trends Genet. 2007;23:243–249. - PubMed
- Du T, Zamore PD. Development (Cambridge, UK) 2005;132:4645–4652. - PubMed
- Bartel DP. Cell. 2004;116:281–297. - PubMed
- Song L, Tuan RS. Birth Defects Res. 2006;78:140–149. - PubMed
Publication types
MeSH terms
Substances
Grants and funding
- DA06668/DA/NIDA NIH HHS/United States
- R37 HD012304/HD/NICHD NIH HHS/United States
- F31 DA021062/DA/NIDA NIH HHS/United States
- R37 HD12304/HD/NICHD NIH HHS/United States
- R03 DA022226/DA/NIDA NIH HHS/United States
- DA022226/DA/NIDA NIH HHS/United States
- R01 DA006668/DA/NIDA NIH HHS/United States
- R37 DA006668/DA/NIDA NIH HHS/United States
LinkOut - more resources
Full Text Sources
Other Literature Sources
Molecular Biology Databases
Research Materials