Interaction of sperm with endometrium can regulate genes involved in endometrial receptivity pathway in mice: An experimental study


Background: Many researchers consider implantation and endometrial receptivity as pertinent issues in reproductive science. Although, several experiments have been performed and their results evaluated, yet there is no confirmed evidence about the related factors and the role of sperm in endometrial receptivity. Objective: To investigate the effect of the sperm-endometrium interaction in regulating genes involved in the endometrial receptivity pathway.

Materials and Methods: In this experimental study, 10 male and 30 female NMRI mice were included, and half of the male cases were vasectomized. The subjects were divided into two groups as follows; group 1 (case) comprised of 15 females mated with 5 non-vasectomized male mice, while group 2 (control) consisted of 15 females mated with 5 vasectomized males. Cases were sacrificed and assessed after 36 hr and the endometrial tissue was extracted and kept at -80°C until the next use. The expression of the endometrial receptivity pathway genes, including VEGF, HBEGF, FGF2, EGF, LIF, LIFR, HOXA10, MUC1, PGR, and CSF, was examined in both groups. For statistical analysis, an independent samples test (Mean ± SD) was used.

Results: The mRNA levels of LIF (p = 0.045), LIFR (p = 0.040), MUC1 (p = 0.032), VEGF (p = 0.022), EFG (p = 0.035), and FGF2 (p = 0.040) were significantly upregulated in the case group compared with the control group.

Conclusion: Finally, seminal plasma was observed to be effective in expressing the involved genes in the successful implantation pathway, including LIF, LIFR, MUC1, VEGF, EGF, and FGF2.

Key words: Endometrial receptivity, Sperm, Gene expression, Mice.

[1] Özatika O, Mungan T, Dag I, Musmuld A. The effect of sperm activation on pinopod formation in endometrial epithelium. J Anat Soc India 2016; 65: S5–S10.

[2] Rincón A, Bolumar D, Valbuena D, Simón C. Use of Molecular markers of endometrial receptivity. In: Rizk B, Khalaf Y. Controversies in assisted reproduction. USA: CRC Press; 2020.

[3] Neykova K, Tosto V, Giardina I, Tsibizova V, Vakrilov G. Endometrial receptivity and pregnancy outcome. J Matern Fetal Neonatal Med 2020; 2: 1–15.

[4] Ajdary M, Keyhanfar F, Aflatoonian R, Amani A, Amjadi F, Zandieh Z, et al. Design and evaluation of a novel nanodrug delivery system for reducing the side effects of clomiphene citrate on endometrium. DARU J Pharm Sci 2020; 28: 1–10.

[5] Ajdary M, Farzan S, Razavi Y, Arabdolatabadi A, Haghparast A. Effects of morphine on serum reproductive hormone levels and the expression of genes involved in fertility-related pathways in male rats. Iran J Pharm Res 2019; 19: 1–26.

[6] Lee SK, Kim CJ, Kim DJ, Kang JH. Immune cells in the female reproductive tract. Immune Netw 2015; 15: 16–26.

[7] Ibrahim LA, Rizo JA, Fontes PLP, Lamb GC, Bromfield JJ. Seminal plasma modulates expression of endometrial inflammatory meditators in the bovine. Biol Reprod 2019; 100: 660–671.

[8] Berger C, Boggavarapu NR, Menezes J, Lalitkumar PGL, Gemzell-Danielsson K. Effects of ulipristal acetate on human embryo attachment and endometrial cell gene expression in an in vitro co-culture system. Hum Reprod 2015; 30: 800–811.

[9] Robertson SA. Seminal plasma and male factor signalling in the female reproductive tract. Cell Tissue Res 2005; 322: 43–52.

[10] He B, Ni Zl, Kong SB, Lu JH, Wang HB. Homeobox genes for embryo implantation: From mouse to human. Anim Model Exp Med 2018; 1: 14–22.

[11] Namiki T, Ito J, Kashiwazaki N. Molecular mechanisms of embryonic implantation in mammals: Lessons from the gene manipulation of mice. Reprod Med Biol 2018; 17: 331– 342.

[12] Yuan J, Deng W, Cha J, Sun X, Borg JP, Dey SK. Tridimensional visualization reveals direct communication between the embryo and glands critical for implantation. Nat Commun 2018; 9: 603–615.

[13] Ribatti D, Tamma R. The chick embryo chorioallantoic membrane as an in vivo experimental model to study human neuroblastoma. J Cell Physiol 2018; 234: 152–157.

[14] Fermin LM, Pain SJ, Morel PCH, Gedye KR, Kenyon PR, Blair HT. Effect of exogenous progesterone on embryo size and ewe uterine gene expression in an ovine ‘dam size’model of maternal constraint. Reprod Fertil Dev 2018; 30: 766–778.

[15] Camargo-Díaz F, García V, Ocampo-Bárcenas A, González-Marquez H, López-Bayghen E. Colony stimulating factor-1 and leukemia inhibitor factor expression from current-cycle cannula isolated endometrial cells are associated with increased endometrial receptivity and pregnancy. BMC Women’s Health 2017; 17: 63–69.

[16] Miah AG, Salma U, Hamano K, Schellander K. Physiological roles of relaxin in prefertilizing activities of spermatozoa. Anim Reprod Sci 2015; 161: 1–15.

[17] Artimani T, Karimi J, Mehdizadeh M, Yavangi M, Khanlarzadeh E, Ghorbani M, et al. Evaluation of prooxidant- antioxidant balance (PAB) and its association with inflammatory cytokines in polycystic ovary syndrome (PCOS). Gynecol Endocrinol 2018; 34: 148–152.

[18] Chelongar R, Hajihosseinlo A, Ajdary M. The effect of Igf-1 and pit-1 genes polymorphisms on fat-tail measurements (fat-tail dimensions) in Makooei sheep. Adv Environ Biol 2014; 8: 862–868.

[19] Robertson SA, Ingman WV, O’Leary S, Sharkey DJ, Tremellen KP. Transforming growth factor β-a mediator of immune deviation in seminal plasma. J Reprod Immunol 2002; 57: 109–128.

[20] Cheng J, Rosario G, Cohen TV, Hu J, Stewart CL. Tissuespecific ablation of the LIF receptor in the murine uterine epithelium results in implantation failure. Endocrinology 2017; 158: 1916–1928.

[21] Chen J, Zhao X, Ao L, Yin T, Yang J. Transcriptomic changes and potential regulatory mechanism of intrauterine human chorionic gonadotropin co-cultured with peripheral blood mononuclear cells infusion in mice with embryonic implantation dysfunction. Ann Transl Med 2020; 8: 99–113.

[22] Shim SH, Kim JO, Jeon YJ, An HJ, Lee HA, Kim JH, et al. Association between vascular endothelial growth factor promoter polymorphisms and the risk of recurrent implantation failure. Exp Ther Med 2018; 15: 2109–2119.

[23] Yue L, Yu HF, Yang ZQ, Tian XC, Zheng LW, Guo B. Egr2 mediates the differentiation of mouse uterine stromal cells responsiveness to HB−EGF during decidualization. J Exp Zoo B Mol Dev Evol 2018; 330: 215–224.

[24] Hu M, Zhang Y, Feng J, Xu X, Zhang J, Zhao W, et al. Uterine progesterone signaling is a target for metformin therapy in PCOS-like rats. J Endocrinol 2018; 237: 123–137.

[25] Altmäe S, Martinez-Conejero JA, Salumets A, Simon C, Horcajadas JA, Stavreus-Evers A. Endometrial gene expression analysis at the time of embryo implantation in women with unexplained infertility. Mol Hum Reprod 2010; 16: 178–187.

[26] Hamlett WC, Musick JA, Hysell CK, Sever DM. Uterine epithelial−sperm interaction, endometrial cycle and sperm storage in the terminal zone of the oviducal gland in the placental smoothhound, Mustelus canis. J Exp Zool 2002; 292: 129–144.

[27] Zandieh Z, Ashrafi M, Aflatoonian K, Aflatoonian R. Human sperm DNA damage has an effect on immunological interaction between spermatozoa and fallopian tube. Andrology 2019; 7: 228–234.

[28] Robertson SA, O’Leary S, Armstrong DT. Influence of semen on inflammatory modulators of embryo implantation. Soc Reprod Fertil Suppl 2006; 62: 231–245.

[29] Elweza AE, Ezz MA, Acosta TJ, Talukder AK, Shimizu T, Hayakawa H, et al. A proinflammatory response of bovine endometrial epithelial cells to active sperm in vitro. Mol Reprod Dev 2018; 85: 215–226.

[30] Reeve L, Ledger WL, Pacey AA. Does the Arg−Gly−Asp (RGD) adhesion sequence play a role in mediating sperm interaction with the human endosalpinx? Hum Reprod 2003; 18: 1461–1468.

[31] Gomes GM, Crespilho AM, Leão KM, Jacob JCF, Gomes LPM, Segabinazzi LG, et al. Can sperm selection, inseminating dose, and artificial insemination technique influence endometrial inflammatory response in mares? J Equine Vet Sci 2019; 73: 43–47.

[32] Bahar L, Kahraman S, Akkuş M, Baykal T. Fine structure and immunohistochemical evaluation of endometrium in fertile and infertile women with implantation failure. Dicle Tıp Dergisi 2012; 39: 269–275.

[33] Burnstock G. Purinergic signalling in the reproductive system in health and disease. Purinergic Signal 2014; 10: 157–187.

[34] Cakmak H, Taylor HS. Implantation failure: molecular mechanisms and clinical treatment. Hum Reprod Update 2011; 17: 242–253.

[35] Kaczmarek MM, Krawczynski K, Blitek A, Kiewisz J, Schams D, Ziecik AJ. Seminal plasma affects prostaglandin synthesis in the porcine oviduct. Theriogenology 2010; 74: 1207–1220.

[36] Gutsche S, Von Wolff M, Strowitzki T, Thaler CJ. Seminal plasma induces mRNA expression of IL-1beta, IL-6 and LIF in endometrial epithelial cells in vitro. Mol Hum Reprod 2003; 9: 785–791.

[37] Alvarez-Rodriguez M, Atikuzzaman M, Venhoranta H, Wright D, Rodriguez-Martinez H. Expression of immune regulatory genes in the porcine internal genital tract is differentially triggered by spermatozoa and seminal plasma. Int J Mol Sci 2019; 20: 513–532.

[38] Carp HJ, Serr DM, Mashiach S, Nebel L. Influence of insemination on the implantation of transferred rat blastocysts. Gynecol Obstet Invest 1984; 18: 194–198.

[39] Maxwell WM, Evans G, Mortimer ST, Gillan L, Gellatly ES, McPhie CA. Normal fertility in ewes after cervical insemination with frozen-thawed spermatozoa supplemented with seminal plasma. Reprod Fertil Dev 1999; 11: 123–126.

[40] Bellinge BS, Copeland CM, Thomas TD, Mazzucchelli RE, O’Neil G, Cohen MJ. The influence of patient insemination on the implantation rate in an in vitro fertilization and embryo transfer program. Fertil Steril 1986; 46: 252–256.

[41] Cicinelli E, De Ziegler D. Transvaginal progesterone: evidence for a new functional’portal system’flowing from the vagina to the uterus. Hum Reprod Update 1999; 5: 365–372.

[42] Pelzer ES, Huygens F, Beagley KW. Steroid hormone dependent inflammation and regulation in the endometrium in women with dysfunctional menstrual cycles: Is there a role for toll-like receptor activation via PAMPs and DAMPs? J Microb Biochem Technol 2016; 8: 344–357.

[43] Wira CR, Fahey JV, Sentman CL, Pioli PA, Shen L. Innate and adaptive immunity in female genital tract: cellular responses and interactions. Immunol Rev 2005; 206: 306–335.