Changes in surface morphology, lectin staining, and gene expression of caprine endometrium exposed to estradiol, progesterone, and mifepristone in vitro

Document Type : Full paper (Original article)

Authors

1 Ph.D. Student in Reproductive Physiology, Department of Zoology, Kurukshetra University, Kurukshetra, India

2 Department of Zoology, Kurukshetra University, Kurukshetra, India

Abstract

Background: In mammalian females, the uterine tissue is highly responsive to steroid hormones and their antagonists. Aims: In the present study, topographical, histoarchitectural, and gene expression changes in goat endometrium treated with estradiol, progesterone, and mifepristone for 24 h were investigated, in vitro. Methods: Scanning electron microscopy (SEM) was used for surface topographical analysis; WGA and DBA lectins were used for histochemical analysis; and qRT-PCR was done for the quantification of mRNA levels of MKI67 (marker of proliferation Ki67), ESR1 (estrogen receptor), PGR (progesterone receptor), CASP3 (caspase 3), and PDGFR-β (platelet derived growth factor receptor-β). Results: Few topographical alterations were observed in endometrial glands and the presence of scattered mucoid granules. A significant decline in WGA staining was reported only in the progesterone group. However, DBA binding was highest in the progesterone group and lowest in the mifepristone group. The expression of MKI67 gene declined to 79% in the mifepristone group, while in the estradiol and progesterone groups it elevated to 153% and 41%, respectively, than control; a similar trend was observed for PDGFR-β. The mRNA abundance for ESR1 declined to 59% in the progesterone group and 10% in the mifepristone group. However, a 100% increase occurred in the estradiol group. PGR expression followed the same trend as that of ESR1. CASP3 declined in the estradiol (50%) and progesterone (37%) group, but it showed a 67% increase in the mifepristone group. Conclusion: We concluded that the caprine uterus undergoes dramatic alteration in structure and functions in response to different kinds of steroidal environments.

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Bailey, DW; Dunlap, KA; Frank, JW; Erikson, DW; White, BG; Bazer, FW; Burghardt, RC and Johnson, GA (2010). Effects of long-term progesterone on developmental and functional aspects of porcine uterine epithelia and vasculature: progesterone alone does not support development of uterine glands comparable to that of pregnancy. Reproduction. 140: 583-594.
Bartel, C; Tichy, A; Schoenkypl, S; Aurich, C and Walter, I (2013). Effects of steroid hormones on differentiated glandular epithelial and stromal cells in a three dimensional cell culture model of the canine endometrium. BMC Vet. Res., 9: 1-16.
Benbia, S; Belkhiri, Y and Yahia, M (2020). Apoptotic cell death in ewe endometrium during the oestrous cycle. J. Hell. Vet. Med. Soc., 71: 2323-2330.
Bookout, AL; Jeong, Y; Downes, M; Yu, RT; Evans, RM and Mangelsdorf, DJ (2006). Anatomical profiling of nuclear receptor expression reveals a hierarchical transcriptional network. Cell. 126: 789-799.
Casanas-Roux, F; Nisolle, M; Marbiax, E; Smets, M; Bassil, S and Donnez, J (1996). Morphometric, immuno-histological and threedimensional evaluation of the endometrium of menopausal women treated by oestrogen and Crinone, a new slow-release vaginal progesterone. Hum. Repro., 11: 357-363.
Chegini, NASSER; Rossi, MJ and Masterson, BJ (1992). Platelet-derived growth factor (PDGF), epidermal growth factor (EGF), and EGF and PDGF beta-receptors in human endometrial tissue: localization and in vitro action. Endocrinology. 130: 2373-2385.
Chen, Y; Wang, W; Zhuang, Y; Chen, X and Huang, L (2011). Effects of low-dose mifepristone administration in two different 14-day regimens on the menstrual cycle and endometrial development: a randomized controlled trial. Contraception. 84: 64-70.
Dahmoun, M; Boman, K; Cajander, S; Westin, P and Backstrom, T (1999). Apoptosis, proliferation, and sex hormone receptors in superficial parts of human endometrium at the end of the secretory phase. J. Clin. Endocrino. Meta. 84: 1737-1743.
D’Amelio, M; Sheng, M and Cecconi, F (2012). Caspase-3 in the central nervous system: beyond apoptosis. Trend. Neurosci., 35: 700-709.
Danielsson, KG; Marions, L and Bygdeman, M (2003). Effects of mifepristone on endometrial receptivity. Steroids. 68: 1069-1075.
De Cock, H; Vermeirsch, H; Ducatelle, R and De Schepper, J (1997). Immunohistochemical analysis of estrogen receptors in cysticendometritis-pyometra complex in the bitch. Theriogenology. 48: 1035-1047.
Dlamini, BJ; Li, Y and Anderson, LL (1995). Mifepristone (RU 486) induces parturition in primiparous beef heifers and reduces incidence of dystocia. J. Ani. Sci., 73: 3421-3426.
Elkabes, S and Nicot, AB (2014). Sex steroids and neuroprotection in spinal cord injury: a review of preclinical investigations. Exp. Neurol., 259: 28-37.
Evans, GS; Gibson, DF; Roberts, SA; Hind, TM and Potten, CS (1990). Proliferative changes in the genital tissue of female mice during the oestrous cycle. Cel. Tiss. Kinet., 23: 619-635.
Ferenczy, A; Bertrand, G and Gelfand, MM (1982) Proliferation kinetics of human endometrium during the normal menstrual cycle. Am. J. Obstet. Gynecol., 133: 859 867.
Fiscella, J; Bonfiglio, T; Winters, P; Eisinger, SH and Fiscella, K (2011). Distinguishing features of endometrial pathology after exposure to the progesterone receptor modulator mifepristone. Hum. Patho., 42: 947-953.
Gerstenberg, C; Allen, WR and Stewart, F (1999). Cell proliferation patterns in the equine endometrium throughout the non-pregnant reproductive cycle. J. Repro. Fert., 116: 167-175.
Gogny, A and Fieni, F (2016). Aglepristone: A review on its clinical use in animals. Theriogenology. 85: 555-566.
Henriet, P; Chevronnay, HPG and Marbaix, E (2012). The endocrine and paracrine control of menstruation. Mol. Cell. Endocrinol., 358: 197-207.
Ing, NH; Spencer, TE and Bazer, FW (1996). Estrogen enhances endometrial estrogen receptor gene expression by a posttranscriptional mechanism in the ovariectomized ewe. Bio. Repro., 54: 591-599.
Jabbour, HN; Kelly, RW; Fraser, HM and Critchley, HO (2006). Endocrine regulation of menstruation. Endocr. Rev., 27: 17-46.
Jang, JH; Woo, SM; Um, HJ; Park, EJ; Min, KJ; Lee, TJ; Kim, SH; Choi, YH and Kwon, TK (2013). RU486, a glucocorticoid receptor antagonist, induces apoptosis in U937 human lymphoma cells through reduction in mitochondrial membrane potential and activation of p38 MAPK. Oncol. Rep., 30: 506-512.
Jones, CJ; Fazleabas, AT; McGinlay, PB and Aplin, JD (1998). Cyclic modulation of epithelial glycosylation in human and baboon (Papio anubis) endometrium demonstrated by the binding of the agglutinin from Dolichos biflorus. Bio. Repro., 58: 20-27.
Klentzeris, LD; Bulmer, JN; Li, TC; Morrison, L; Warren, A and Cooke, MID (1991). Lectin binding of endometrium in women with unexplained infertility. Fert. Ster., 56: 660-667.
Linde-Forsberg, C; Kindahl, H and Madej, A (1992). Termination of mid-term pregnancy in the dog with oral RU 486. J. Sma. Ani. Pract., 33: 331-336.
Maentausta, O; Svalander, P; Danielsson, KG; Bygdeman, M and Vihko, R (1993). The effects of an antiprogestin, mifepristone, and an antiestrogen, tamoxifen, on endometrial 17 beta-hydroxysteroid dehydrogenase and progestin and estrogen receptors during the luteal phase of the menstrual cycle: an immunohistochemical study. J. Clinic. Endocrino. Metabol., 77: 913-918.
Monroe, DG; Berger, RR and Sanders, MM (2002). Tissue-protective effects of estrogen involve regulation of caspase gene expression. Mol. Endocrino., 16: 1322-1331.
Moorthy, K; Yadav, UC; Siddiqui, MR; Sharma, D; Basir, SF and Baquer, NZ (2004). Effect of estradiol and progesterone treatment on carbohydrate metabolizing enzymes in tissues of aging female rats. Biogerontology. 5: 249-259.
Motta, PM and Andrews, PM (1976). Scanning electron microscopy of the endometrium during the secretory phase. J. Anat., 122: 315-322.
Munson, L; Kao, JJ and Schlafer, DH (1989). Characterization of glycoconjugates in the bovine endometrium and chorion by lectin histochemistry. Reproduction. 87: 509-517.
Narvekar, N; Cameron, S; Critchley, HO; Lin, S; Cheng, L and Baird, DT (2004). Low-dose mifepristone inhibits endometrial proliferation and up-regulates androgen receptor. J. Clin. Endocrino. Metab., 89: 2491-2497.
Newton, GR; Lewis, SK; Avendano, J; Williams, EA; Ribeiro, FRB; Nuti, LC; Foxworth, WB and Ing, NH (2019). Fucosyltransferase gene expression in goat endometrium during the estrous cycle and early pregnancy. Theriogenology. 132: 118-127.
Peralta, LE; Olarte, MR; Arganaraz, M; Ciocca, D and Miceli, DC (2005). Progesterone receptors: their localization, binding activity and expression in the pig oviduct during follicular and luteal phases. Domest. Anim. Endocrinol., 28: 74-84.
Rodriguez-Martinez, H; Persson, E; Hurst, M and Stanchev, P (1992). Immunohistochemical localization of platelet-derived growth factor receptors in the porcine uterus during the oestrous cycle and pregnancy. J. Vet. Med. Ser. A., 39: 1-10.
Sant’Ana, FJF; Nascimento, EF; Andres Laube, PF; Gimeno, EJ and Barbeito, CG (2009). Lectin-binding sites on the normal and pathologic uterus of sows. Repro. Domes. Ani., 44: 889-893.
Sharma, AK and Sharma, RK (2020). Effect of prostaglandins E2 and F2α on granulosa cell apoptosis in goat ovarian follicles. Iran. J. Vet. Res., 21: 97-102.
Sharma, RK and Singh, P (2021). Histomorphometric analysis of goat uterine tissue on in vitro exposure with ovarian hormones and mifepristone. Bulg. J. Vet. Med., 24: 12-21.
Spornitz, UM; Socin, CD and Dravid, AA (1999). Estrous stage determination in rats by means of scanning electron microscopic images of uterine surface epithelium. Anat. Rec., 254: 116-126.
Szymanska, M and Blitek, A (2016). Endometrial and conceptus response to exogenous progesterone treatment in early pregnant gilts following hormonally-induced estrus. Ani. Repro. Sci., 174: 56-64.
Tian, YZ; Liu, YP; Tian, SC; Ge, SY; Wu, YJ and Zhang, BL (2020). Antitumor activity of ginsenoside Rd in gastric cancer via up-regulation of Caspase-3 and Caspase-9. Die Pharmaz., 75: 147-150.
Tokashiki, S; Kawashima, Y; Sugimura, M and Kudo, N (1976). Histochemical changes in the endometrium of the goats during the course of estrous cycle and pregnancy (author’s transl). Nihon juigaku zasshi. Japan. J. Vet. Sci., 38: 639-645.
Vermeirsch, H; Van den Broeck, W; Coryn, M and Simoens, P (2002). Immunohistochemical detection of androgen receptors in the canine uterus throughout the
estrus cycle. Theriogenology. 57: 2203-2216.
Verstegen-Onclin, K and Verstegen, J (2008). Endocrinology of pregnancy in the dog: a review. Theriogenology. 70: 291-299.
Vierbuchen, M (1991). Lectin receptors. In: Berry, CL and Grundmann, E (Eds.), Current topics in pathology. London, UK, Springer. 83: 271-361.
Wagenfeld, A; Saunders, PT; Whitaker, L and Critchley, HO (2016). Selective progesterone receptor modulators (SPRMs): progesterone receptor action, mode of action on the endometrium and treatment options in gynecological therapies. Exp. Opin. Therap. Targ., 20: 1045-1054.
Walter, I and Bavdek, S (1997). Lectin binding patterns of porcine oviduct mucosa and endometrium during the oestrous cycle. J. Anat., 190: 299-307.
Wang, Y; Jiang, X and Wang, S (2014). The influence of mifepristone to caspase 3 expression in adenomyosis. Clinic. Experi. Obstet. Gyneco., 41: 154-157.
Yanaihara, A; Otsuka, Y; Iwasaki, S; Aida, T; Tachikawa, T; Irie, T and Okai, T (2005). Differences in gene expression in the proliferative human endometrium. Fert. Ster., 83: 1206-1215.