Retrospective analysis of day-5 euploid and aneuploid blastocoel fluid apoptotic gene expression.MATERIALS AND METHODS: Blastocoel fluid-conditioned media (25mL) was collected following trophectoderm (TE) biopsy of ICSI-generated day-5 blastocysts. Biopsied TE cells were sent for preimplantation genetic testing for aneuploidies using NGS. The blastocoel-fluid conditioned media from 12 embryos were each subjected to DNase I treatment prior to cDNA synthesis before assessing gene expression via RT-PCR using TaqMan Fast Array-Human Apoptosis plates (assessing 92 apoptosis associated genes).RESULTS: Of the 92 apoptotic genes analyzed, NOD1, NOD2 and TRADD expression were detected only in aneuploid embryos compatible with life. CASP5 gene expression was detected only in autosomal aneuploidies (n¼4 Down syndrome and n¼1 Edwards syndrome), while LTB was detected only in sex chromosome aneuploidies (n¼1 Klinefelter syndrome and n¼2 Turner syndrome). Aneuploid embryos (n¼3) incompatible with life did not reveal any unique gene expression profiles. The euploid embryo (resulting in a term birth) analyzed did not show expression of any of the aforementioned genes.CONCLUSIONS: This is the first report of gene expression differences detected in blastocoel fluid between survivable and non-survivable aneuploidies and an euploid embryo resulting in a term birth. While differences in apoptotic gene expression were observed among the embryos analyzed, we saw no similarities between an euploid embryo and survivable aneuploidies. Interestingly, we detected CASP5 and LTB expression unique to autosomal aneuploidies and sex chromosomal aneuploidies, respectively. Further studies will elucidate the processes by which certain aneuploidies can result in a live birth.
OBJECTIVE: We performed this study to investigate the expression of class 1, 2, 3, 4 and 5 histone deacetylase (HDAC-1, 2, 3, 4, and 5) mRNA and proteins in eutopic and ectopic endometrial tissues of patients with endometriosis and in eutopic endometrial tissues of women without endometriosis, and to evaluate the relationship between HDAC and the development of endometriosis. DESIGN: Prospective cohort study. MATERIALS AND METHODS: For this prospective cohort study, twenty patients who underwent surgery for stage III or IV endometriosis for the study group and 20 patients who underwent surgery for other benign gynecologic disease for controls between Jul 2010 to Mar 2013. All subjects were not pregnant and had normal regular menstruation. No one received hormonal therapy for at least 6 months before surgical treatment. During surgical treatment, eutopic endometrial tissues were collected from study and control groups and ectopic endometrial tissues were collected from study group. To quantify the expression of HDAC mRNA, real time reverse transcriptase polymerase chain reaction (RT-PCR) was employed. To measure the expression of HDAC protein, Western blotting was employed. Mean values were expressed as meanAEstandard deviation (SD). Kruskal-Wallis test was used to compare the mean value. Statistical significance was defined as P<0.05. All analyses were performed by using SPSS statistical package for Windows, version 11.0 (SPSS Inc, Chicago, IL). RESULTS: The amounts of HDAC-1, 2, 3, 4, and 5 mRNAs were significantly higher in eutopic and ectopic endometrial tissue of patients with endometriosis compared to eutopic endometrial tissue of normal women (p < 0.05). The expression of HDAC 1 protein was significantly higher in eutopic and ectopic endometrial tissue of patients with endometriosis. CONCLUSIONS: Our results suggest the possible relationship between the development ofendometriosis and HDAC. Especially high level of HDAC 1 in endometrial tissue may be associated with the development of endometriosis. References: 1. Eleftherios P. Samartzis, MD1, Aurelia Noske, et al. The Expression of histone deacetylase 1, but not other class I histone deacetylases, is significantly increased in endometriosis.
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