Luteinizing hormone-induced connexin 43 down-regulation: inhibition of translation.

The coordinated function of the different compartments of the follicle, the oocyte and the somatic cumulus/granulosa cells, is enabled by the presence of a network of cell-to-cell communication generated by gap junctions. Connexin 43 (Cx43) is the most abundant gap junction protein expressed by the ovarian follicle. The expression of Cx43 is subjected to the control of gonadotropins as follows: FSH up-regulates, whereas LH down-regulates its levels. The aim of this study was to explore the mechanism by which LH reduces the levels of Cx43 and to identify the signal transduction pathway involved in this process. The effect of LH was studied in vitro using isolated intact ovarian follicles. The possible mediators of LH-induced Cx43 down-regulation were examined by incubating the follicles with LH in the presence or absence of inhibitors of protein kinase A (PKA) and of MAPK signaling pathways. Our experiments revealed a 3-h half-life of Cx43 in both control and LH-treated follicles, suggesting that LH did not affect the rate of Cx43 degradation. We further demonstrated that the level of Cx43 mRNA was not significantly influenced by this gonadotropin. However, upon LH administration, [(35)S]methionine incorporation into Cx43 protein was remarkably reduced. The LH-induced arrest of Cx43 synthesis was counteracted by inhibitors of both the PKA and the MAPK cascades. We show herein that LH inhibits Cx43 expression by reducing its rate of translation and that this effect is mediated by both PKA and MAPK.

Local injury to the endometrium doubles the incidence of successful pregnancies in patients undergoing in vitro fertilization.

OBJECTIVE: Exploration of the possibility that local injury of the endometrium increases the incidence of implantation. DESIGN: Prospective study. SETTING: Clinical IVF unit. PATIENT(S): A group of 134 patients, defined as good responders to hormonal stimulation, who failed to conceive during one or more cycles of IVF and embryo transfer (ET). INTERVENTION(S): The IVF treatment and ET were preceded by repeated endometrial biopsies, in a randomly selected 45 of a total of 134 patients. MAIN OUTCOME MEASURES: Outcome of IVF-ET treatments. RESULT(S): Transfer of a similar number of embryos (3.4 +/- 1.0 and 3.1 +/- 0.9 in the experimental and control patients, respectively) resulted in rates of implantation (27.7% vs. 14.2%, P =.00011), clinical pregnancy (66.7% vs. 30.3%, P =.00009), and live births per ET (48.9% vs. 22.5%, P =.016) that were more than twofold higher in the experimental group as compared to controls. CONCLUSION(S): These results suggest that IVF treatment that is preceded by endometrial biopsy doubles the chance for a take-home baby.

The Israeli National Committee for sex selection by pre-implantation genetic diagnosis: a novel approach (2005-2011).

BACKGROUND: Pre-implantation genetic diagnosis (PGD) for fetal sex selection raises complex dilemmas. In Israel, PGD is regulated by the Ministry of Health. It is basically prohibited, but exceptions can be made upon approval by the National Committee for Sex Selection by PGD for Non-Medical Reasons (the "Committee"). This report describes the Committee's work since its inception in May, 2005 through December, 2011. METHODS: Files were abstracted onto a structured form. Discrete variables were analyzed by chi-square analysis, and continuous variables by T-Test. RESULTS: During the study period 411 applications were received. Two-thirds of the applicants (n = 276; 67.2%) were Jewish and 26.8% were Moslem Arab. Over two-thirds (n = 285; 69.3%) had no children of the requested sex and ≥4 children of the opposite sex. Three-quarters of the requests were for a male (n = 308; 74.9%): 100% of Arab and 63% of Jewish applicants. Many noted more than one reason for their request. The most frequent category (n = 201; 48.9%) was a strong emotional desire, followed by medically-related reasons (n = 83; 20.2%). For 216 applications a decision was arrived at, with 46 (21.3%) approved. Of the remaining 195 for 192 over a year had passed since last contact with the Committee. The likelihood of approval was higher if applicants met the criterion of ≥4 same-sex children than if they didn't (33.7% vs. 11.6%, P = 0.001). The largest number of approvals were those requested for 'emotional' reasons, while the highest approval rate was for religious reasons. CONCLUSIONS: This study reviewed the first seven years of Committee activity. Most requested males, and the primary reason was the parents' intense emotional desire. Only one-fifth of the decisions were approvals, possibly reflecting reluctance to encourage non-medically-indicated PGD, a viewpoint not unique to Israel. Limitations include the relatively small number of cases and lack of access to Committee deliberation protocols. It is recommended that longitudinal studies be conducted to gain insight into the consequences to individuals, couples and families--both those whose requests were approved and those denied-- of this major step in reproductive technologies and in society's effort to respond to them.

Local injury of the endometrium induces an inflammatory response that promotes successful implantation.

OBJECTIVE: To study whether an injury-induced inflammation might be the mechanism underlying the favorable effect of endometrial biopsy on the implantation rate in in vitro fertilization (IVF) patients. DESIGN: Controlled clinical study. SETTING: A medical center IVF unit and a research institute. PATIENT(S): Women undergoing IVF who had previous failed treatment cycles. INTERVENTION(S): Endometrial samples were collected from two groups of patients on day 21 of their spontaneous menstrual cycle. The experimental, but not the control group underwent prior biopsy treatment on days 8 or/and 11 to 13 of that same cycle. MAIN OUTCOME MEASURE(S): Abundance of immune cells, cytokines/chemokines level, correlation between these parameters and pregnancy outcome. RESULT(S): A statistically significantly higher amount of macrophages/dendritic cells (HLA-DR+ CD11c+ cells) and elevated proinflammatory cytokines, tumor necrosis factor-α (TNF-α), growth-regulated oncogene-α (GRO-α), interleukin-15 (IL-15), and macrophage inflammatory protein 1B (MIP-1B), were detected in day-21 endometrial samples of the experimental group. A direct stimulatory effect of TNF-α on MIP-1B, GRO-α, and IL-15 messenger RNA (mRNA) expression was demonstrated. A positive correlation was found between the levels of macrophages/dendritic cells, MIP-1B expression, and TNF-α expression and the pregnancy outcome. CONCLUSION(S): A biopsy-induced inflammatory response may facilitate the preparation of the endometrium for implantation. Increased MIP-1B expression could possibly serve for prediction of implantation competence.

Successful pregnancy and delivery of a healthy baby after endometrial biopsy treatment in an in vitro fertilization patient with severe Asherman syndrome.

OBJECTIVE: To implement the procedure of endometrial biopsy in a case of severe Asherman syndrome as a possible treatment to increase uterine receptivity. DESIGN: Case report. SETTING: IVF Unit, Kaplan Medical Center, Rehovot, Israel. PATIENT(S): A 29-year-old patient with severe Asherman syndrome, who underwent six operative hysteroscopies combined with hormonal treatment, and no functional receptive endometrium was achieved. INTERVENTION(S): We performed three endometrial biopsies on days 8, 12, and 21 of a progyluton-induced menstrual cycle, and a fourth biopsy on day 21 of the next induced menstrual cycle. After that cycle the patient underwent an IVF treatment. MAIN OUTCOME MEASURE(S): Ultrasound measurement of endometrial thickness, serum beta-hCG, sonography test for the presence of a gestational sac with heartbeat, and pregnancy follow-up until birth. RESULT(S): Biopsy treatment increased the thickness of the endometrium from unobservable by sonography to 7 mm on the day of hCG administration. The next IVF cycle resulted in implantation of an embryo and the birth of a healthy baby boy. CONCLUSION(S): Repeated endometrial biopsies may be used in patients with Asherman syndrome immediately after forming a uterine cavity by hysteroscopy to improve its receptivity.

Endometrial biopsy-induced gene modulation: first evidence for the expression of bladder-transmembranal uroplakin Ib in human endometrium.

OBJECTIVE: To explore the possibility that endometrial injury modulates the expression of specific genes that may increase uterine receptivity. DESIGN: Controlled clinical study. SETTING: Clinical IVF unit and academic research center. PATIENT(S): IVF patients with 28- to 30-day menstrual cycles. INTERVENTION(S): Endometrial biopsies from two groups of patients were collected on days 20-21 of their spontaneous menstrual cycle. The experimental, but not the control, group underwent biopsies on days 11-13 and 21-24 of their preceding cycle. MAIN OUTCOME MEASURE(S): Global endometrial gene expression and specific analysis of uroplakin Ib (UPIb) mRNA level throughout the menstrual cycle. RESULT(S): Local injury modulated the expression of a wide variety of genes. One of the prominently up-regulated genes was the bladder transmembranal protein, UPIb, whose expression by the endometrium is shown here for the first time. Endometrial UPIb mRNA increases after biopsy in the same cycle wct 2with an additional elevation in the following cycle. Immunohistochemical analysis localized the UPIb protein to the glandular-epithelial cells. Genes encoding other membrane proteins such as adipose differentiation-related protein and mucin 1, transmembrane, were also up-regulated. CONCLUSION(S): The biopsy-induced increase in the expression of UPIb and other genes encoding membrane proteins supports the possible importance of the membrane structure and stability during implantation. The specific role of UPIb in uterine receptivity should be elucidated.

Connexin43 in rat oocytes: developmental modulation of its phosphorylation.

It is well established that the 43-kDa connexin (Cx43) is predominantly expressed by ovarian somatic cells, whereas the identity of the connexins contributed by the oocyte to form gap junctions with its neighboring cells is not fully elucidated. Our study aimed to examine oocytes for the expression and regulation of Cx43 throughout oogenesis. Growing and fully grown rat oocytes that were meiotically incompetent and competent, respectively, were examined. Fully grown oocytes were analyzed either before or after reinitiation of meiosis as well as at the second meiotic metaphase. Immunofluorescent analysis of zona pellucida-free oocytes using conventional and confocal microscopy demonstrated a characteristic pattern of punctuated staining of Cx43 on the oolema. Immunogold electron microscopy localized Cx43 to the oocyte surface and the microvillar processes. Reverse transcriptase-polymerase chain reaction and Western blot analysis revealed similar amounts of Cx43 gene and protein in oocytes of different developmental stages. However, a relative increase in the phosphorylated forms of the protein was observed in fully grown oocytes that had completed their maturation. Our findings demonstrate that rat oocytes express a developmentally regulated Cx43. They further suggest that homotypic gap junctions that consist of Cx43 may be present between rat oocytes and their adjacent cumulus cells.