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National Scientific and Technical Research Council of Argentina (CONICET), Argentina
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National Scientific and Technical Research Council of Argentina (CONICET), Argentina
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Laboratory of Descriptive, Comparative and Experimental Histology and Embriology (LHYEDEC). Faculty of Veterinary Science, National University of La Plata (UNLP), Argentina
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In the present work, we emphasize the studies about integrins and their receptors in pig placental interface at different times of gestation. Uterine placental interface (n = 24) of 17, 30, 60 and 70 days of gestation (dg) and non-pregnant uterus (n = 4) of crossbred sows were used. The presence of αvβ3 (ITGAV (integrin subunit alpha V) ITGB3 (integrin subunit beta 3)) and α5β1 (ITGA5 (integrin subunit alpha V) ITGB3 (integrin subunit beta 3)) integrins, and their ligands fibronectin (FN) and osteopontin (OPN/ SPP1), was detected by immunohistochemistry, and the immunolabeled area percentage (IAP) and the optical density (OD) were determined. The integrins and its ligands analyzed have presented peaks of expression in early and mid-gestation, both in IAP and in the OD area, decreasing at 70 dg. These temporal changes showed us that the molecules studied in this work participate in embryo/feto–maternal attachment, variably. Besides, we found a significant correlation both in the intensity and in the extension of immunostaining for trophoblastic FN and endometrial αvβ3, and trophoblastic OPN and endometrial α5β1, throughout the entire pig pregnancy. At late gestation, there is notable placental remodeling with subsequent removal or renewal of folds at the uterine–placental interface that results in the loss of focal adhesions. The decrease of the expression of some integrins and their ligands in late gestation, particularly at 70 dg, would demonstrate that there would be other adhesion molecules and other ligands that could be participating in the establishment of the maternal–fetal interface.
Lay summary
To carry a successful pregnancy, the formation of the placenta in pigs depends on adhesion molecules. Some of these molecules called integrins bind to other molecules such as fibronectin (FN) and osteopontin (OPN/SPP1). The variation in those molecule amounts during gestation would indicate which molecule is participating and what role it plays in pregnancy. We worked with pig placentas of early, mid- and late- gestation and non-pregnant uteruses. αvβ3 (ITGAV (integrin subunit alpha V) ITGB3 (integrin subunit beta 3)) and α5β1 (ITGA5 (integrin subunit alpha 5) ITGB1 (integrin subunit beta 1)) integrins, FN and OPN were found until mid-gestation but not at late gestation, suggesting that other types of molecules have a role in the last period of gestation.
Biological Sciences Department, College of Science, King Faisal University, Al Ahsa, Saudi Arabia
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Follicular fluid (FF), a product of vascular transudate and granulosa and thecal cell secretions, is the milieu that has evolved to support oocyte growth and maturation which plays a central role in oocyte quality determination. Therefore, a suboptimal FF composition may be reflected in compromised oocyte progression through maturation, fertilization, or embryo development. To date, the composition of bovine FF remains understudied. To address this, we comprehensively characterized the metabolomic constituency of bovine FF in the period during which the oocyte undergoes meiotic maturation. More specifically, FF from pre (−24 h) and peri (−2 h)-ovulatory follicles was profiled by high-throughput untargeted ultra-HPLC tandem mass spectroscopy. A total of 634 metabolites were identified, comprising lipids (37.1%), amino acids (30.0%), xenobiotics (11.5%), nucleotides (6.8%), carbohydrates (4.4%), cofactors and vitamins (4.4%), peptides (3.6%), and energy substrates (2.1%). The concentrations of 67 metabolites were significantly affected by the stage of follicle development, 33.3% (n = 21) were reduced (P ≤ 0.05) by a mean of 9.0-fold, whereas 46 were elevated (P ≤ 0.05) by a mean of 1.7-fold in peri- vs pre-ovulatory FF. The most pronounced individual metabolite concentration decreases were observed in hypoxanthine (98.9-fold), xanthine (65.7-fold), 17β-oestradiol (12.4-fold), and inosine (4.6-fold). In contrast, the greatest increases were in retinal (4.9-fold), 1-methyl-5-imidazoleacetate (2.7-fold), and isovalerylcarnitine (2.7-fold). This global metabolomic analysis of bovine FF temporal dynamics provides new information for understanding the environment supporting oocyte maturation and facilitating ovulation that has the potential for improving oocyte quality both invivo and in vitro.
Lay Summary
The ovaries are part of the female reproductive system, and they produce and store eggs in structures known as ‘follicles’. Depending on the species, one or more follicles release an egg from the ovary during ovulation. FF, which is formed from the secretions of follicle cells and substances delivered from the bloodstream, bathes the eggs as they develop within their follicles. For pregnancy to happen, the egg must be capable of being fertilised by a sperm cell, developing into an embryo and implanting it in the womb. FF has evolved to support the egg to achieve this. Using the cow as a model, this study looks at the composition of FF during the final hours before ovulation, when the egg becomes mature and ready for fertilisation. More than 600 different substances were identified, providing new information, that has the potential to improve egg quality.
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Kisspeptin, a hypothalamic neuropeptide encoded by the KISS1 gene, has a pivotal role in promoting gonadotrophin-releasing hormone secretion in mammals. Kisspeptin and its receptor (KISS1R) are also expressed in certain peripheral tissues including gonads, suggesting intra-gonadal roles. Such actions at the level of the bovine ovary have not been explored previously. The current aims were to determine whether KISS1 and KISS1R are expressed in the bovine ovary and whether kisspeptin or a kisspeptin antagonist can modulate ovarian steroid production by cultured ovarian cells. Granulosa cells (GC) and theca interna cells (TC) were collected from antral follicles (3–18 mm) categorized into five class sizes. Early, mid and regressing corpora lutea (CL) were also collected for RT-qPCR analysis of KISS1 and KISS1R expression. Bovine TC and GC cultured under both non-luteinizing (serum-free) and luteinizing (serum-supplemented) conditions were treated for 4 days with kisspeptin-10 (10– 10–10– 6M) or kisspeptin antagonist (kp234; 10–10–10–6M), alone and in combination with either follicle-stimulating hormone (GC), luteinizing hormone (TC) or forskolin (luteinized GC/TC). Steroid secretion (GC: oestradiol, progesterone; TC: androstenedione, progesterone; luteinized GC/TC: progesterone) was measured by ELISA and viable cell number determined by neutral red uptake assay. KISS1 and KISS1R transcripts were detected in TC, GC and CL with significant differences between follicle categories and CL stages. However, neither kisspeptin-10 nor kisspeptin antagonist affected steroid secretion or viable cell number in any of the four ovarian cell culture models. As such, the hypothesis that kisspeptin has a direct intraovarian role to modulate follicular or luteal steroidogenesis, or cell proliferation/survival, is not supported.
Lay summary
Kisspeptin-producing nerve cells (neurones) in the hypothalamus play a crucial role in controlling the reproductive system. Kisspeptin activates receptors on gonadotrophin-releasing hormone neurones which, in turn, stimulate the pituitary gland to release gonadotrophins. Gonadotrophins act on the gonads (ovaries or testes) to regulate their function (e.g. ovarian follicle development and steroid production). Evidence has emerged in several species that kisspeptin and its receptor are also present in certain peripheral tissues, including the ovaries, suggesting ‘local’ actions. So far, few studies have investigated this. Here, we first show that both kisspeptin and its receptor are expressed by key ovarian cell types of cattle. However, we found that treating cultured bovine theca and granulosa cells with kisspeptin or kisspeptin antagonist did not modify steroid secretion. As such, the hypothesis that kisspeptin has a direct intraovarian role to modulate ovarian steroid production is not supported.
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Treatment of sub-fertile women aged ≥ 40 years old (advanced maternal age (AMA)) is challenging. Co-treatment with growth hormone (GH) is suggested to improve reproductive outcomes in poor responders. However, few studies, and with conflicting results, focused on women of AMA. A systematic review and meta-analysis of randomized controlled trials (RCTs) and comparative retrospective trials (CRTs) of GH cotreatment in AMA women undergoing in vitro fertilization or intracytoplasmic injection treatment using their autologous oocytes was performed. The search included studies published in English up to the end of 2021. The primary outcome was the clinical pregnancy rate per embryo transfer. Secondary outcomes were the number of mature and retrieved oocytes and the rate of live birth. A total of 406 studies were found. The final analysis included 3 RCTs and 4 CRTs with 481 patients who used GH and 400 patients who did not. Clinical pregnancy and live birth rates were significantly higher in the GH cotreatment group compared to the placebo as well as the group without GH co-treatment, (odds ratio (OR): 2.2; 95% CI: 1.34–3.61 and OR: 4.12; 95% CI: 1.82–9.32, respectively). Intriguingly, the subgroup analysis showed that poor-responder patients did not benefit from co-treatment with GH. There were no statistically significant differences in the number of mature or retrieved oocytes. GH cotreatment in a subgroup of women of AMA improves clinical pregnancy and live birth per fresh embryo transfer. However, this conclusion must be taken with caution and further research is needed. The review is registered in the PROSPERO database (www.crd.york.ac.uk/prospero/; CRD42021252618).
Lay summary
Women over 40 years undergoing in vitro fertilization (IVF) treatment commonly require high doses of injectable medications to stimulate their ovaries. Co-treatment with growth hormone (GH) has been shown to enhance the ovarian response and improve the outcome. The investigators found seven studies that compared 881 women over 40 years of age who had undergone IVF treatment with or without GH cotreatment. Statistical analysis of data from these studies showed that some of these women may benefit from adding a GH to their ovarian stimulation medications. The benefit was evident in those with good ovarian reserve. Women over 40 years with a good ovarian reserve can increase their chance of pregnancy by 4–20% when using GH during ovarian stimulation. However, this finding requires confirmation in a well-designed study with large sample size. Furthermore, the optimal dose, regimen, safety, and cost-effectiveness of GH cotreatment should be clarified.
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The University of Edinburgh, Edinburgh, UK
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Lay Summary
During the COVID-19 pandemic, the public delayed seeking medical help, which may have affected the impact of having an ectopic pregnancy. An ectopic pregnancy is when pregnancy tissue grows outside its normal position in the womb, and it can be life-threatening. It can be treated by non-surgical or surgical options, and any delay in seeking help can reduce the options for treatment and increase the need for more urgent management. We wanted to assess whether there were differences in the presentation and management of ectopic pregnancies in a major teaching hospital between 2019 (pre-COVID-19) and 2021 (COVID-19 period). We found that the pandemic did not cause a delay in seeking medical help or cause worse outcomes. In fact, immediate surgical treatment and time in the hospital were less during COVID-19, perhaps because of a desire to avoid admission to hospital. One outcome of COVID-19 is reassurance that we can safely use more non-surgical treatments for ectopic pregnancies.
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Lay summary
Men and boys with cancer treated with chemotherapy are known to have reduced fertility following their treatment. This is because some chemotherapy drugs can damage the cells in the testicles that make sperm. This study found there is limited information available on the effect of one group of chemotherapy drugs, called taxanes, on testicular function and fertility. More studies are needed to aid clinicians in advising patients on how this taxane-based chemotherapy may affect their future fertility.
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Department of Clinical Andrology, Centre of Reproductive Medicine and Andrology, Muenster, Germany
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Objective
Germ cells of transwomen are affected by gender-affirming hormone therapy (GAHT). Fertility will be lost after surgical intervention; thereby, fertility preservation becomes an increasingly imortant topic. This study investigated if the absolute number of spermatogonia in transwomen is comparable at the time of gender-affirming surgery (GAS) to that in pre-pubertal boys.
Methods
We carried out a retrospective study of testicular tissues from 25 selected subjects, which had undergone a comparable sex hormone therapy regimen using cyproterone acetate (10 or 12.5 mg) and estrogens. As controls, testicular biopsies of five cisgender adult men (aged 35–48 years) and five pre-/pubertal boys (5–14 years) were included. Testicular tissues were immunohistochemically stained for MAGE A4-positive cells, the most advanced germ cell type. The number of spermatogonia per area was assessed. Clinical values and serum hormone values for FSH, LH, testosterone, free testosterone, estradiol and prolactin were determined on the day of GAS for correlation analyses.
Results
Round spermatids were the most advanced germ cell type in 3 subjects, 5 had an arrest at spermatocyte stage, while 17 showed a spermatogonial arrest. On average, testicular tissues of transwomen contained 25.15 spermatogonia/mm3, a number that was significantly reduced compared to the two control groups (P < 0.01, adult 80.65 spermatogonia/mm3 and pre-/pubertal boys 78.55 spermatogonia/mm3). Linear regression analysis revealed that testes with higher weight and high LH contained more spermatogonia.
Conclusion
Irrespective of treatment dose or duration, spermatogenesis was impaired. Spermatogonial numbers were significantly reduced in transwomen compared to the control groups.
Lay summary
When transwomen go through treatment to confirm their gender, their germ cells are affected. They lose their fertility after surgery, so fertility preservation becomes an important topic. We carried out a study looking at tissue from testes of 25 people who had been through the same sex hormone therapy until surgery. Blood samples were also taken. As controls, samples were taken from the testes of cisgender boys and adult men. On average, the samples from the testes of transwomen contained a smaller number of early sperm cells compared to the two control groups. Regardless of the dose or length of hormone treatment, the fertility of transwomen was significantly reduced so that counseling about fertility preservation should be offered before hormone therapy.
Academic Department of Obstetrics and Gynaecology, Level 9 Worsley Building, School of Medicine, University of Leeds, Leeds, UK
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Lay summary
Friction caused by blood flowing across cells that line blood vessels (endothelial cells) activates sensors of mechanical force. This produces nitric oxide (NO) which widens placental blood vessels, enabling more blood flow to the baby. This study sought to determine whether the mechanical sensor, Piezo1, is important for NO production in fetoplacental endothelial cells (FpECs) and whether the steps in this pathway are different in small for gestational age (SGA) babies, where placental blood flow is often altered. We showed that in healthy FpECs, blood flow increased NO signalling. We suggest that in SGA babies, FpECs have an increase in baseline levels of NO signalling, suggestive of a compensatory drive. Treating healthy and SGA cells with a Piezo1 chemical activator, Yoda1, upregulated NO signalling. This shows that Piezo1 is linked to NO and that in SGA, FpECs have the capacity to further increase NO. Further research will establish whether Piezo1 enhancement leads to increased blood flow in the placenta. If so, Piezo1 could be a new target for developing treatments to prevent poor growth of babies in the womb.
Fertigo Medical Ltd., Zichron Yaakov, Israel
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The Felsenstein Medical Research Center, the Sackler Faculty of Medicine, Tel-Aviv University
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Adelson School of Medicine, Ariel University, Ariel, Israel
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Graphical abstract
Abstract
Background
The distribution of the blood vessel network at any point in time in any body tissue may provide valuable information with regard to the tissue condition, whether it is in a growth, declining or recovery phase as well as giving insights as to its angiogenesis functionality. The blood vessel three-dimensional network of the endometrium goes through a process of change over a relatively short period of 4 weeks on average. It is well accepted that angiogenesis within the endometrium is closely related to the success or failure of the implantation of the embryo.
Objective and rationale
Our study aims to present a method to follow the three-dimensional evolution of the superficial blood vessel distribution in the endometrium throughout the uterine cycle.
Method
This method utilizes differences in the observed broadband colors of the blood vessels in order to assess their depth coordinate below the endometrial tissue surface. We implemented the method using microscopic images of fresh, ex vivo, endometrial samples of different cycle days to obtain the statistical evolution track of the superficial blood vessel population in both human and animal (swine) samples.
Outcomes
In human samples, we observed a systematic and consistent trend in the blood vessel diameter distribution at different tissue depths. We demonstrate that the magnitude of this trend evolves throughout the course of the female cycle.
Wider implications
This method has the potential to further our understanding of the mechanisms of angiogenesis in tissues other than the endometrium. We propose that this method may also contribute to more precise endometrial dating and may assist in more accurate determination of embryo transfer timing within in vitro fertilization treatments.
Lay summary
The inner lining tissue of the womb (uterus) is called the endometrium, and it undergoes significant changes during the menstrual cycle.
The endometrium blood vessel network goes through rapid changes during the cycle.
We have developed a new method to measure this through surface imaging of the endometrium.
We use samples of endometrial tissues collected at different dates in the cycle to show how useful this method is in evaluating the development of the endometrium.
The method may also be used to investigate different processes of generating new blood vessels and may help to support dating the development of the endometrium.
Our work offers a non-invasive or minimally invasive method which reveals the three-dimensional blood vessel network and may be used to help in a variety of diagnoses. For example, this method may be used to see how receptive the uterus is during in vitro fertilization treatment.
