Biologie de la Reproduction, Environnement, Epigénétique et Développement

There is no ideal treatment for Peyronie's Disease (PD) since every patient has different needs with varying perceived risks versus benefits of individual treatment options, coupled with the degree of impact relating to PD on sexual function and beyond. The 5th International Consultation on Sexual Medicine (ICSM) guideline on PD provides an update to the previous ICSM with an aim to provide a clinical framework and best-evidenced consensus recommendations to assist clinicians in managing PD. Available literature was reviewed, and a modified Delphi consensus was undertaken by leading sexual medicine experts to achieve a set of Quality of Evidence and strength of recommendations. This guideline was developed as an authoritative best-evidenced consensus incorporating state-of-the-art knowledge and contemporary literature. A total of thirty-one recommendations were made on PD guidelines. New updates to the 4th ICSM on PD have been made in definition, pathophysiology, and treatment strategies. The presence of penile pain with erection could occur in the chronic phase due to torque or stretch on the penile scar and surrounding tissue. Oral therapy works best during the acute phase and should be provided concurrently with other treatment modalities. Penile traction therapy can reduce penile deformity as monotherapy or may be offered as part of a multimodal therapy approach. There is no convincing evidence supporting cellular-based therapy. The type of plication procedure used depends on the surgeon and patient preference, although excision of the tunica followed by plication may have a higher complication rate. There is currently no ideal graft, and graft contracture can occur postoperatively. Adjunctive procedures such as penile modeling, tunical plication, plaque incision/excision and grafting can be performed when penile deformity and/or penile curvature persist following penile prosthesis implantation. The management of PD should be individualized based on the patient's needs and preferences while balancing the benefits and risks of each treatment option.

Abstract Variants with intermediate functional effects—those that neither eliminate nor fully maintain normal gene function—represent an underrecognized source of genetic complexity and constitute a grey zone that challenges variant classification. Here, we address this issue using GT>GC (+2T>C) 5′ splice-site variants as a tractable model, as approximately 15% of such substitutions retain variable amounts of wild-type (WT) transcript. Using residual WT transcript as a quantitative readout, we first show that disease-associated GT>GC variants known to generate substantial WT transcript (for example, SPINK1 c.194+2T>C, HBB c.315+2T>C, and BRCA2 c.8331+2T>C) consistently complicate classification efforts. We then carried out a locus-wide assessment of all 26 GT>GC substitutions in CFTR , applying SpliceAI delta donor-loss scores as an initial screen and cross-checking classifications in expert-curated database. This evaluation indicated that three variants with low or intermediate SpliceAI scores are likely to retain WT transcript. Minigene analyses of four selected CFTR variants, together with additional examination of a BAP1 GT>GC variant with conflicting clinical interpretations, illustrate the methodological challenges in predicting and experimentally quantifying residual WT transcript. These results emphasize that (i) in silico predictors cannot reliably discriminate between complete and partial splice disruption and (ii) experimental systems differ in their ability to detect subtle splicing outcomes. Overall, our integrative analysis shows that GT>GC variants generating appreciable WT transcript exemplify a broader class of intermediate-effect alleles that complicate both computational prediction and experimental assessment, underscoring the need for classification frameworks that incorporate quantitative functional data and better capture the continuum of variant effects.

Understanding how ageing impacts endometrial function is crucial for preserving fertility in older females. While ageing-related cellular dysfunction and inflammation are observed in the bovine uterus, its effects on endometrial physiology remain unclear. Using an experimental model of young (4-7 years) and old (13-15 years) cloned female cattle, we assessed the effect of ageing on the endometrium through transcriptome profiling and responses of cultured endometrial cells to interferon tau (IFNT) and of cultured endometrial explants to LPS. Progesterone profiles were similar between young and old females. Transcriptomic analysis of endometrial biopsies on day 15 of the estrous cycle identified 859 differentially expressed genes (DEG; p ≤ 0.05, |FC| ≥ 1.5), among which 402 DEG were over-expressed and 457 DEG were under-expresssed in old females. These DEG are linked to immune, inflammatory, metabolic, and cell organization pathways and networks. RT-qPCR validation of selected candidate DEG revealed an increased expression of COL4A3, CPA3, IGFBP1, IGFBP2, RSAD2, SCARA5, and SERPINA14 in young females. In vitro stimulation with IFNT of primary uterine glandular epithelial and stromal cells revealed that glandular epithelial cells exhibit a greater sensitivity to IFNT than stromal cells, both in old and young females. Glandular epithelial cells derived from old females exhibit a weaker response to IFNT, in terms of the number of differentially expressed genes, compared to those from young females. The effect of LPS treatment on cytokine concentrations was lightly more pronounced in young females than in old females, with LPS leading to a significant increase in the concentration of IL-1α, IL-1β, IL-6 and IL-10 in young females. By altering the transcriptomic profile of the endometrium and its capacity to respond to both the embryo's signal and inflammatory factors, we propose that age may be a key factor underlying uterine-related reproductive failures. Further experiments are required to confirm this hypothesis.

Pluripotent stem cells have long been used to produce knockout mice via germline chimera technology. However, aside from the rat, this approach has not been successfully applied to other mammals. Here, we demonstrate that rabbit induced pluripotent stem cells (iPSCs) can be reprogrammed using KLF2, ERAS and PRMT6, enabling them to efficiently colonize embryos. These chimeric embryos can develop into fetuses and newborn rabbits, with iPSCs contributing up to 100 % to certain organs. Notably, female rabbits generated through this method are healthy and transmit the iPSC genome to their offspring with a high efficiency, demonstrating germline chimerism. This advancement establishes a foundation for developing rabbit models of human disease with complex genetic traits.

Feeding levels can vary in goat farms for several reasons. Underfeeding of gestating dairy goats can occur and may influence fetal follicle development and future reproductive performance in their offspring. The objective of this experiment was to study the effect of restricted feeding during the last third of gestation on some reproductive parameters in female offspring (F1). Two feeding groups were formed using 60 Alpine and Saanen dairy goats to produce the F1 female offspring. The control group (C, n = 30) was fed to requirements. The restricted group (R, n = 30) was given the same diet, but the quantity corresponded to 50% of the amount given to the C group between -8 and -4 weeks, 60% between -5 and -4 weeks, 70% between -4 and -3 weeks and 80% from -2 weeks to parturition. Estrus was synchronized at 7 months of age in female F1 goats born to C (n = 17) or born to R goats (n = 15) with a progestagenimpregnated sponge and prostaglandin F2α and eCG injections, and the goats were inseminated. Serial blood samples were collected over this period. After mating, plasma progesterone rose more slowly to reach a maximum plateau concentration in females born to R goats compared to females born to C goats (P < 0.05). The lag time was approximately 2 days. There was no difference in dam F0 prolificacy (C, 2.4 ± 0.6 vs. R, 2.1 ± 0.8, P > 0.05), length of gestation (C, 151 ± 2 days vs. R, 151 ± 2 days) and kid mortality rate (mummified fetuses, stillbirth or death in first 48h) between the groups. However, birth weight was lower in R kids compared to C kids (4.0 ± 0.11 kg vs. 4.5 ± 0.10 kg, P = 0.007). In conclusion, maternal feed restriction during late pregnancy modified progesterone patterns after insemination in female offspring, although there was no effect on reproductive success.

Sperm samples are separated into bad and good quality samples in function of their phenotype, but this does not indicate their genetic quality. Here, we used GeneChip miRNA arrays to analyze microRNA expression in ten semen samples selected based on high-magnification morphology (score 6 vs. score 0) to identify miRNAs linked to sperm phenotype. We found 86 upregulated and 21 downregulated miRNAs in good-quality sperm (score 6) compared with bad-quality sperm samples (score 0) (fold change > 2 and p-value < 0.05). MiR-34 (FC × 30, p = 8.43 × 10-8), miR-30 (FC × 12, p = 3.75 × 10-6), miR-122 (FC × 8, p = 0.0031), miR-20 (FC × 5.6, p = 0.0223), miR-182 (FC × 4.83, p = 0.0008) and miR-191 (FC × 4, p = 1.61 × 10-6) were among these upregulated miRNAs. In silico prediction algorithms predicted that miRNAs upregulated in good-quality sperm targeted 910 genes involved in key biological functions of spermatozoa, such as cell death and survival, cellular movement, molecular transport, response to stimuli, metabolism, and the regulation of oxidative stress. Genes deregulated in bad-quality sperm were involved in cell growth and proliferation. This study reveals that miRNA profiling may provide potential biomarkers of sperm quality.

ABSTRACTPurposeThe influence of culture media used during in vitro fertilization (IVF) on offspring phenotype remains controversial. However, specific effects of short exposure time after fertilization remain underexplored. By evaluating Ca2+ oscillations as a readout of the first response of eggs to their microenvironment, we aim to investigate if early differences correlate with later adult phenotypes.MethodsOocytes fertilized by ICSI were cultured for 4 h in three different media (Cook and Vitrolife, used in human IVF, and KSOM, used for mouse embryos). They were either measured for Ca2+ oscillations or transferred into pseudo‐pregnant females. After birth, growth curves of pups were measured up to adulthood and various organs were weighed.ResultsCulture media significantly modulate Ca2+ oscillations during oocyte activation. ICSI‐fertilized oocytes cultured in Cook and Vitrolife exhibited fewer oscillations, lower frequency, and reduced variability compared to KSOM. These early differences correlated with long‐term developmental outcomes: females from Cook and Vitrolife cultures were heavier throughout growth and had larger adult organ sizes compared to those from KSOM.ConclusionsBrief exposure to media immediately after ICSI shapes Ca2+ dynamics and adult phenotypes. Optimizing embryo culture protocols in assisted reproductive technologies may improve IVF outcomes by modulating metabolic pathways linked to development.