Transforming Growth Factor-β2 and Its Receptor Type II Messenger RNA Levels in Mice Endometrium and Their Regulation by Sex Steroids During Estrous Cycle
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Despite increasing evidence pointing to the involvement of transforming growth factor-β superfamily in the regulation of reproduction, a definitive role of TGF-β2 and its receptor type II in the uterus is currently unclear. In this study, using semi-quantitative reverse transcriptase PCR (RT-PCR) and real-time PCR techniques (qPCR), TGF-β2 and its receptor type II (TGF-βRII) mRNA levels were examined in mouse endometrium during estrous cycle and in response to an acute exposure of 17β-estradiol (E2) and progesterone (P4). We found that TGF-βRII mRNA was expressed at a very low level during diestrus, and remained comparatively higher at proestrus, estrus and metestrus stages with no significant differences between them. On the contrary, the average TGF-β2 mRNA levels abruptly increased from estrus to diestrus stages and then briskly decreased at proestrus, the proliferative stage of the cycle. These results suggest that both TGF-β2 and its receptor type II are functional during proliferation as well as apoptosis of mice endometrial epithelial and stromal cells. A single injection of E2 or P4 at diestrus stage mice caused significant attenuation of the expression of TGF-β2 mRNA after 8 h of treatment compared to sesame oil control. Inhibition induced by P4 was comparatively stronger than that induced by E2 and results further showed that combined injection of E2 and P4 had similar inhibitory effects as that of P4 alone on TGF-β2 expression. In contrast, expression of TGF-βRII mRNA was not significantly altered by ovarian steroids administered either alone or in combination. Results further demonstrated that in short-term endometrial tissue incubation, p38 MAPK pathway was involved in the steroid-mediated inhibition of TGF-β2 mRNA with no effect on TGF-βRII mRNA. This result suggests that p38 MAPK pathway may be an important regulator of TGF-β2 and TGF-βRII mRNA expression in vitro in diestrus endometrium, in cooperation with ovarian steroids.
KeywordsTGF-β2 TGF-βRII Estrogen Progesterone Estrous cycle Mouse uterus
This work is supported by grant-in-aid given to the first author Payel Guha as DST-INSPIRE Fellowship (Reg. No. IF- 131055) and by Department of Science and Technology (DST), Government of India (Ref. No. SR/SO/BB-0116/2012) and partially by UGC SAP to Department of Zoology, Kalyani University. We would like to thank Department of Microbiology, Department of Molecular Biology and Biotechnology, University of Kalyani for various infrastructural supports.
Payel Guha- First author- full contribution; Other authors –partial contribution.
Compliance with Ethical Standards
Conflict of interest
The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.
- Ando, N., F. Hirahara, J. Fukushima, S. Kawamoto, K. Okuda, T. Funabashi, I. Gorai, and H. Minaguchi. 1998. Differential gene expression of the TGF-beta isoforms and TGF-beta receptors during the the first trimester of pregnancy at the human maternal-fetal interface. American Journal of Reproductive Immunology 40: 48–56.CrossRefPubMedGoogle Scholar
- Bustin, S.A., V. Benes, J.A. Garson, J. Hellemans, J. Huggett, M. Kubista, R. Mueller, T. Nolan, M.W. Pfaffl, G.L. Shipley, and J. Vandesompele. 2009. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clinical Chemistry 55: 611–622.CrossRefPubMedGoogle Scholar
- Caron, P.L., G. Frechette-Frigon, C. Shooner, V. Leblanc, and E. Asselin. 2009. Transforming growth factor beta isoforms regulation of Akt activity and XIAP levels in rat endometrium during estrous cycle, in a model of pseudopregnancy and in cultured decidual cells. Reproductive Biology and Endocrinology 7: 80–93.CrossRefPubMedGoogle Scholar
- Casslen, B., T. Sandberg, B. Gustavsson, R. Willen, and M. Nilbert. 1998. Transforming growth factor β1 in the human endometrium. Cyclic variation, increased expression by estradiol and progesterone, and regulation of plasminogen activators and plasminogen activator inhibitor-1. Biology of Reproduction 58: 1343–1350.CrossRefPubMedGoogle Scholar
- Chegini, N.A.S.S.E.R., Y.O.N.G. Zhao, R.S. Williams, and K.C. Flanders. 1994. Human uterine tissue throughout the menstrual cycle expresses transforming growth factor-beta 1 (TGF beta 1), TGF beta 2, TGF beta 3, and TGF beta type II receptor messenger ribonucleic acid and protein and contains [125I] TGF beta 1-binding sites. Endocrinology 135: 439–449.CrossRefPubMedGoogle Scholar
- Gold, L.I., B. Saxena, K.R. Mittal, M. Marmor, S. Goswami, L. Nactigal, M. Korc, and R.I. Demopoulos. 1994. Increased expression of transforming growth factor β isoforms and basic fibroblast growth factor in complex hyperplasia and adenocarcinoma of the endometrium: evidence for paracrine and autocrine action. Cancer Research 54: 2347–2358.PubMedGoogle Scholar
- Gupta, A., C.M. Dekaney, F.W. Bazer, M.M. Madrigal, and L.A. Jaeger. 1998. Beta transforming growth factors (TGFβ) at the porcine conceptus–maternal interface. Part II: uterine TGFβ bioactivity and expression of immunoreactive TGFβs (TGFβ1, TGFβ2, and TGFβ3) and their receptors (type I and type II). Biology of Reproduction 59: 911–917.CrossRefPubMedGoogle Scholar
- Lea, R.G., K.C. Flanders, C.B. Harley, J. Manuel, D. Banwatt, and D.A. Clark. 1992. Release of a tranforming growth factor (TGF)-β2-related suppressor factor from postimplantation murine decidual tissue can be correlated with the detection of a subpopulation of cells containing RNA for TGF-β2. Journal of Immunology 148: 778–787.Google Scholar
- Marshburn, P.B., A.M. Arici, and M.L. Casey. 1994. Expression of transforming growth factor-β1 messenger ribonucleic acid and the modulation of deoxyribonucleic acid synthesis by transforming growth factor-β1 in human endometrial cells. American Journal of Obstetrics and Gynecology 170: 1152–1158.CrossRefPubMedGoogle Scholar
- Massague, J., and Y.G. Chen. 2000. Controlling TGF-β signaling. Genes & Development 14: 627–644.Google Scholar
- Paul, S., K. Pramanick, S. Kundu, D. Kumar, and D. Mukherjee. 2010. Regulation of ovarian steroidogenesis in vitro by IGF-I and insulin in common carp, Cyprinus carpio: stimulation of aromatase activity and P450arom gene expression. Molecular and Cellular Endocrinology 315: 95–103.CrossRefPubMedGoogle Scholar
- Sachdeva, G., V. Patil, R.R. Katkam, D.D. Manjramkar, S.D. Kholkute, and C.P. Puri. 2001. Expression profiles of endometrial leukemia inhibitory factor, transforming growth factor β2 (TGFβ2), and TGFβ2 receptor in infertile bonnet monkeys. Biology of Reproduction 65: 1–8.CrossRefPubMedGoogle Scholar
- Sellheyer, K., J.R. Bickenbazh, J.A. Rothnagel, D. Bundman, M.A. Longley, T. Krieg, N.S. Roche, A.B. Roberts, and D.R. Roop. 1993. Inhibition of skin development by overexpression of transforming growth factor βl in epidermis of transgenic mice. Proceedings of National Academy of Sciences, USA 90: 5237–5241.CrossRefGoogle Scholar
- Shull, M.M., I. Ormsby, A.B. Kier, S. Pawlowski, R.J. Diebold, M. Yin, R. Allen, C. Sidman, G. Proetzel, D. Calvin, N. Annunziata, and T. Doetschman. 1992. Targeted disruption of the mouse transforming growth factor-βl gene results in multifocal inflammatory disease. Nature 359: 693–699.CrossRefPubMedPubMedCentralGoogle Scholar
- Wada, K., S. Nomura, E. Morii, Y. Kitamura, Y. Nishizawa, A. Miyake, and N. Terada. 1996. Changes in levels of mRNAs of transforming growth factor (TGF)-β1,-β2,-β3, TGF-β type II receptor and sulfated glycoprotein-2 during apoptosis of mouse uterine epithelium. The Journal of Steroid Biochemistry and Molecular Biology 59: 367–375.CrossRefPubMedGoogle Scholar
- Xia, W., M.T. Longaker, and G.P. Yang. 2006. P38 MAP kinase mediates transforming growth factor-β2 transcription in human keloid fibroblasts. American Journal of Physiology: Regulatory, Integrative and Comparative Physiology 290: 501–508.Google Scholar
- Zenclussen, M.L., N. Linzke, A. Schumacher, S. Fest, N. Meyer, P.A. Casalis, and A.C. Zenclussen. 2015. Heme oxygenase-1 is critically involved in placentation, spiral artery remodeling, and blood pressure regulation during murine pregnancy. Frontiers in Pharmacology 5: 291.CrossRefPubMedPubMedCentralGoogle Scholar