RESUMEN
Immunohistochemistry is a suitable method for the detection of proteins from the Claudin family and several antibodies are commercially available for the detection of Claudin congeners. Immunodetection of Caludin-4 in the paraffin-embedded specimens might be a useful tool for studying the role of these proteins in the cyclic transformation of the endometrium and its role in the endometrial receptivity; furthermore, other components of the junctional zone involved in the transformational process of the endometrium can be detected by means of immunohistochemistry/immunofluorescence with several polyclonal or monoclonal antibodies. The aim of this chapter is to comprehensively overview the materials and methods to perform the endometrial biopsy and to detect Claudin-4 in paraffin-embedded samples of endometrium. Additionally, the interpretation of the results is addressed.
Asunto(s)
Endometrio/metabolismo , Inmunohistoquímica/métodos , Proteínas de la Membrana/metabolismo , Biotina , Claudina-4 , Eosina Amarillenta-(YS) , Femenino , Técnica del Anticuerpo Fluorescente/métodos , Hematoxilina , Humanos , Adhesión en Parafina/métodosRESUMEN
To investigate the luteal phase endometrial expression of leukemia inhibitor factor (LIF), insulin-like growth factor 1 (IGF-1), progesterone receptor (PR), claudin 4 (CLDN4), vascular-endothelial growth factor receptor 3 (VEGFR-3), bone morphogenetic protein 4 (BMP-4) and citokeratin 7 (CK-7), we obtained luteal phase endometrial samples from 52 women. Samples were dated and integrated using a tissue microarray (TMA). Samples were immunostained for LIF, IGF-1, PR, CLDN4, VEGFR-3, BMP-4 and CK-7. Frequencies of positive expressions at the early, mid and late luteal phases were compared by two proportions test. Concomitant expression of these proteins was assessed with Chi-square or Fischer's test. The frequency of LIF was positively correlated to the frequency of IGF-1 (r = 0.99; p < 0.05) and PR (r = 0.99; p < 0.05), and the correlation between IGF-1 and PR tended to be significant (r = 0.98; p < 0.1). The expression of PR was associated with the absence of CLDN4 (p < 0.001). Thus, expression of LIF, IGF-1 and PR are correlated during the luteal phase, and immunohistochemistry for these proteins might be used to assist in the assessment of endometrial maturation. In addition, the expression of CLDN4 and PR was not concomitant, warranting further investigation on the relationship of their endometrial expression.
Asunto(s)
Endometrio/metabolismo , Fase Luteínica/metabolismo , Adulto , Proteína Morfogenética Ósea 4/metabolismo , Distribución de Chi-Cuadrado , Claudina-4 , Femenino , Humanos , Inmunohistoquímica , Factor I del Crecimiento Similar a la Insulina/metabolismo , Queratina-7/metabolismo , Factor Inhibidor de Leucemia/metabolismo , Proteínas de la Membrana/metabolismo , Selección de Paciente , Análisis por Matrices de Proteínas , Receptores de Progesterona/metabolismo , Receptor 3 de Factores de Crecimiento Endotelial Vascular/metabolismoRESUMEN
OBJECTIVE: To determine whether gonadotropin releasing hormone (GnRH)-agonist or -antagonist induces higher percentages of cumulus cell apoptosis and if the use of either is detrimental to ART outcomes. PATIENTS: Women in a private facility under treatment for IVF had their cumulus cells isolated and analyzed by flow cytometry. Viable, apoptotic, and dead cumulus cell rates related to ovarian stimulation by GnRH-agonist or -antagonist were measured and compared with fertilization and implantation rates. RESULTS: Treatment with GnRH-agonist produced a greater number of follicles than treatment with GnRH-antagonist. No differences in implantation and pregnancy rates were found. While cumulus cell (CC) apoptosis was positively correlated with estradiol on the day of hCG administration, no significant difference in the percentage of apoptotic cells between treatments was detectable. Additionally, implantation rate and the average follicular estradiol production on the day of hCG administration were no different between treatments. CONCLUSIONS: GnRH-agonist or -antagonist treatment protocols induce similar levels of apoptosis in CCs and are not detrimental to ART outcomes.