LEYLA BAHAR, Semra KAHRAMAN, MURAT AKKUŞ, Tülin BAYKAL

Fertil kadınlar ve implantasyon başarısızlığı olan infertil kadınlarda endometriumun ince yapı ve immünohistokimyasal değerlendirilmesi

Amaç: Endometrial reseptivite; endometrium epitelinin fonksiyonel başarısı olan geçici bir süreçtir. Bu çalışmada, fertil ve Tekrarlayan implantasyon başarısızlığı (TİB) olan bireylerin endometrium dokuları, Transmisyon Elektron Mikroskobuyla (TEM) ve E-cadherinle immünohistokimyasal açıdan değerlendirilmiştir. Amacımız, sadece fertil ve TİB olan kadınların endometrium dokularının karşılaştırılmasını yapmak değil aynı zamanda endometriumda implantasyon sürecinin hücresel çatısı ve mekaniğinin daha iyi anlaşılmasını sağlamaktır. Gereç ve yöntem: Araştırmamız için İn vitro fertilizasyon (İVF)/ Embriyo Transferi (ET) sonrası gebelik oluşmayan onyedi infertil ve on fertil kadın çalışmamıza dahil edilmiştir. Rutin ışık mikroskobu ve TEM teknikleriyle doku takibi sağlanmıştır. Bulgular: Fertil grubun endometrium yüzey epitelinde implantasyon belirteci pinopod oluşumuna yoğun olarak rastlanırken, infertil grupta yetersiz pinopod oluşumu ve belirgin silya ve mikrovillus varlığı dikkat çekmektedir. E-cadherin’le yapılan immünohistokimya çalışmasında skorlanma yapılmıştır. Fertil grup epitelinde immünreaktivite çok belirsizken, infertil grupta daha belirgin bir boyanma olduğu gözlenmiştir. Sonuç: Son bulgular E-cadherin’in reseptif dönemdeki endometrial epitele yapıştırıcı özellikler aktarıyor olması yanısıra, ikili fonksiyona sahip olması üzerinedir. İlk aşamalarda, hücre yüzeyinde yapışkanlık sağladığı, daha sonra epitel hücrelerde ayrılma yaptığı ve blastosistin invazyonunu etkinleştirmek için suprese olduğu düşünülmektedir. Bu çalışmada TİB grubunun aksine, fertil grup epitelinin, çok zayıf boyanması, blastosist implantasyonunu kolaylaştırması açısından önemlidir.

Fine structure and immunohistochemical evaluation of endometrium in fertile and infertile women with implantation failure

Objectives: Being a temporary process, endometrial receptivity is the functional success of endometrial process. In this study, the fertile and recurrent implantation failure (RIF) of individuals with endometrial tissues is analyzed by Transmission Electron Microscopy (TEM) and immunohistochemistry using the E-cadherin. Our goal is not only to make a comparison of endometrial tissues in women with RIF and fertile but also to provide a better understanding of the process and mechanics of the cellular roof during the implantation of endometrium. Materials and methods: Our research consists of seventeen infertile women who are after In-Vitro fertilization (IVF) / embryo transfer (ET) not occur on pregnancy and ten fertile women were included. Tissue follow-up were performed using routine light microscopy and TEM techniques. Results: In fertile group, markers of endometrial implantation in the surface epithelium, which are pinopodes were found in heavily. In infertile group, there was prominent cilia formation, microvillus and the presence of inadequate pinopod. E-cadherin using immunohistochemistry study was scoring. When immune-reactivity was very pale in the epithelium of the fertile group, infertile groups showed a clear staining. Conclusion: Recent findings; E-cadherin transmits adhesive properties to receptive period of endometrial epithelium as well as, having dual functions. Firstly, the cell surface adhesion is provided, then the separation of epithelial cells and is thought to be suppressed to enable the invasion of the blastocyst. Unlike TIB group, weak staining of epithelium of the fertile group, is important to facilitate implantation of the blastocyst.

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1. Susan JK. Molecular interactions at the maternal-embryonic interface during the early phase of implantation. Sem Reprod Med 2000;18(3):237-43.
2. Attar E. Spermatogenezis, Fertilizasyon, Erken embriyo gelişimi ve İmplantasyon. Umur Çolgar. Reproduktif Endokrinoloji ve İnfertilite, 1. baskı. İstanbul: İstanbul Medikal Yayıncılık, 2006:34-45.
3. Paria BC, Lim H, Wang XN, Liehr J, Das SK, Dey SK. Coordination of different effects of primary estrogen and catecholestrogen on two distinct targets mediates embryo implantation in the mouse. Endocrinology 1998;139(12):5235-46.
4. Sunder S, Lenton E. Endocrinology of the Peri- Implantation Period. Clin Obstet Gynaecol 2000;14(5):789-800.
5. Murphy CR, Shaw TJ. Plasma membrane transformation: a common response of uterine epithelial cells during the periimplantation period. Cell Biol Int 1994;18(12):1115-28.
6. Murphy CR. Junctional barrier complexes undergo major alterations during the plasma membrane transformation of uterine epithelial cells. Hum Reprod 2000;15(3):182-8.
7. Salmani MK, Nikzad H, Shiokawa S, Akimoto Y, Iwashita M. Secretory role for human uterodomes (pinopods): secretion of LIF. Mol Hum Reprod 2005;11(8):553–9.
8. Rashid NA, Lalitkumar S, Lalitkumar PG, Gemzell-Danielsson K. Endometrial Receptivity and Human Embryo Implantation. Am J Reprod Immunol 2011;66(1):23–30.
9. Defrere S, Langendonck A, Moulin P, et al. Human endometrial epithelial cells (EEC) contitutively express more intercellular adhezion molecule (ICAM)-1 than endometrial stromal cells (ESC) in culture. Am J Reprod Immunol 2005;54(1):5-12.
10. Nikas G, Develioglu OH, Toner JP, Jones HW. Endometrial pinopodes indicate a shift in the window of receptivity in IVF cycles. Hum Reprod 1999;14(3):787–92.
11. Tan BK, Vandekerckhove P, Kennedy R, Keay SD. Investigation and current management of recurrent IVF treatment failure in the UK. BJOG 2005;112(6):773–80.
12. Dey SK, Das SK, Reese J, Paria BC, Daikoku T, Wang H. Molecular cues to implantation. Endocr Rev 2004;25(3):341-73.
13. Lee CN, Chang SW, Cho NH, Cho SH. Nitrous oxide synthase expression in placenta of preeclampsia. J Korean Med Sci 1997;12(6):532-8.
14. Mutlu C, Koyutürk M, Karpuz V. Evaluation of endothelial nitric oxide synthase (eNOS) immunreactivity of fetal and maternal placenta. Cerrahpaşa J Med 2005;36(1):109-15.
15. İltemur DC, Şatıroğlu H, Berker B, Çetinkaya H, Kahraman K. Yardımla üreme tekniklerinde implantasyon ve gebelik oranlarını etkileyen faktörler. T Klin J Gynel Obst 2003;13(4):466-75.
16. Akar M, Kurşun S, Taşkın Ö. Tekrarlayan İn vitro Fertilizasyon Başarısızlıklarında Yaklaşım. Türk Fertil Der 2004;12(3):217-24.
17. Bahar L, Baykal T. İmplantasyon Sürecinde Endometriyum. Mersin Üniversitesi Sağlık Bilimleri Dergisi 2008;1(2):1-7.
18. Ledee-Bataille N, Lapree-Delage G, Taupin JL. Concentration of leukaemia inhibitory factor (LIF) in uterine flushing fluid is highly predictive of embryo implantation. Hum Reprod 2002;17(1):213-8.
19. Bee K, Tan BK, Vandekerckhove P, Kennedy R, Keay SD. Investigation and current management of recurrent IVF treatment failure in the UK. Int J O & G 2005;112(6):773- 80.
20. El-Toukhy T. and Taranissi M. Towards better quality research in recurrent implantation failure: standardizing its definition is the first step. Reprod Biomed Online 2006;12(3):383-5.
21. Elder K, Dale B. Preimplantation Genetic Diagnosis. In Vitro Fertilization, 2nd edn. Cambridge: Cambridge University Press, 2000:1-33.
22. Bahar L, Kahraman S, Baykal T. Comparatively Evaluation of Infertile (RIF) and Fertile Women Endometrial Biopsies at the Ultrastructural Level by TEM. In: Dubuisson JB, Gomel V. Reproductive Medicine and Surgery from the Proceedings of the 15th World Congress on In Vitro Fertilization and 4th World Congress on In Vitro Maturation, 1st edn. Geneva, Monduzzi International Proceedings, 2009:171-3.
23. Nikas G. Pinopodes as markers of endometrial receptivity in clinical practice. Hum Reprod 1999;14(2):99-106.
24. Tinel H, Denker HW, Thie M. Calcium influx in human uterine epithelial RL95-2 cells triggers adhesiveness for trophoblast-like cells. Model studies on signalling events during embryo implantation. Mol Hum Reprod 2000;6(12):1119–30.
25. Thie M, Harrach-Ruprecht B, Sauer H, Fuchs P, Albers A, Denker HW. Cell adhesion to the apical pole of epithelium: a function of cell polarity. Eur J Cell Biol 1995;66(2):180–91.
26. Bloorl DJ, Metcalfe AD, Rutherford A. Brison DR. Kimber SJ. Expression of cell adhesion molecules during human preimplantation embryo development. Mol Hum Reprod 2002;8(3): 237-45.
27. Fujimoto J, Ichigo S, Hori M, Tamaya T. Alteration of Ecadherin, alpha- and beta-catenin mRNA expression in human uterine endometrium during the menstrual cycle. Gynecol Endocrinol 1996;10(1):187–91.
28. Dawood MY, Lau M, Khan-Dawood FS. E-cadherin and its Messenger ribonucleic acid in periimplantation phase human endometrium in normal and clomiphene-treated cycles. Am J Obstet Gynecol 1998;178(5):996–1001.
29. Poncelet C, Leblanc M, Walker-Combrouze F, et al. Expression of cadherins and CD44 isoforms in human endometrium and peritoneal endometriosis. Acta Obstet Gynecol Scand 2002;81(3):195–203.
30. Gumbiner B, Stevenson B, Grimaldi A. The role of the cell adhesion molecule uvomorulin in the formation and maintenance of the epithelial junctional complex. J Cell Biol 1988;107(4):1575–87.
31. Li Q, Wang J, Armant DR, Bagchi MK, Bagchi IC. Calcitonin down-regulates E-cadherin expression in rodent uterine epithelium during implantation. J Biol Chem 2002;277(4):447–55.
32. Kumar S, Zhu LJ, Polihronis M, et al. Progesterone induces calcitonin gene expression in human endometrium within the putative window of implantation. J Clin Endocrinol Metab 1998;83(12):4443–50.
33. Zhu LJ, Bove KC, Polihronis M. Calcitonin is a progesterone regulated marker which forecasts the receptive state of endometrium during implantation. Endocr J 1998;139(9):3923–34.
34. Rahnama F, Thompson B, Steiner M, et al. Epigenetic regulation of e-cadherin controls endometrial receptivity. Endocrinology 2009;150(3)1466–72.
35. Shih HC, Shiozawa T, Miyamoto T, et al. Immunohistochemical expression of e-cadherin and catenin in the normal and malignant human endometrium: An inverse correlation between e-cadherin and nuclear -catenin expression. Anticancer Res 2004;24(8):3843-50.