Reproduction

Abad, Cilia; Musilova, Ivana; Cifkova, Eva; Portillo, Ramon; Kumnova, Fiona; Karahoda, Rona; Sterba, Martin; Lisa, Miroslav; Kacerovsky, Marian; Stranik, Jaroslav; Stuchlik, Ales; Staud, Frantisek

doi: 10.1530/rep-24-0378pmid: 40192828

In briefIntrauterine inflammation disrupts tryptophan metabolism in both the placenta and the fetal brain, leading to a shift toward neurotoxic metabolites. These findings highlight the critical role of placental function in neurodevelopment and suggest that inflammation-induced metabolic changes may contribute to neurodevelopmental disorders.AbstractThe placenta plays a crucial role beyond nutrient transfer, acting as a dynamic endocrine organ that significantly influences maternal physiology and fetal development. It responds rapidly to even slight changes in the in utero environment to promote fetal survival. Disruptions in placental function are increasingly recognized as key contributors to the origins of neurodevelopmental disorders. In this study, we employed advanced technology to induce intrauterine inflammation through ultrasound-guided administration of LPS into gestational sacs. We then evaluated its effects on the gene expression of enzymes involved in TRP metabolism and conducted a comprehensive LC/MS analysis of the metabolome in the placenta and fetal brain of Wistar rats. Our results show that intraamniotic injection of LPS induces a robust inflammatory response leading to significant alterations in TRP metabolism, including downregulation of tryptophan hydroxylase (TPH) in the placenta, resulting in a decrease in serotonin (5-HT) levels. Similarly, in the fetal brain, exposure to LPS led to reduced Tph expression and increased monoamine oxidase expression, suggesting a decrease in 5-HT synthesis and an increase in its degradation. Furthermore, an upregulation of the kynurenine pathway was observed in both the placenta and fetal brain. Moreover, we detected a shift toward neurotoxicity, evidenced by an imbalance between neuroprotective and neurotoxic metabolites, including decreased levels of kynurenic acid and upregulation of kynurenine monooxygenase in the fetal brain. In conclusion, our findings reveal significant alterations in TRP metabolism following intrauterine inflammation, potentially contributing to neurodevelopmental disorders.

Gutfreund, Niklas; Tuppi, Marcel; Schäfer, Birgit; Klinger, Francesca Gioia; Kirchhof, Anja; Hofmann, Antonia; Dikic, Ivan; Dötsch, Volker

doi: 10.1530/rep-24-0225pmid: 40226943

In briefInduction of DNA double-strand breaks results in oocyte death caused by the activation of TAp63α. This study investigates which chemotherapeutic drugs activate p63 and which cause oocyte death without p63 activation.AbstractPrimary ovarian insufficiency is a severe side effect of classical chemotherapy and radiotherapy in treatment of female cancer patients of reproductive age. The p53-homolog TAp63α emerged as the key protein regulating apoptosis following DNA damage in oocytes of primordial follicles. In this study, we monitored the toxicity of widely used chemotherapeutic agents on oocytes of primordial follicles utilizing a GFP-c-kit transgenic mouse model, the involvement of active TAp63α and the mechanism of action leading to its activation. Our studies show that alkylating agents and topoisomerase II poisons are potent activators of TAp63α by directly inducing DNA damage. Oxidative stress and DNA intercalation were not sufficient to trigger TAp63α activation despite showing a strong general toxicity. These results are in agreement with several previous investigations that have demonstrated that DNA double-strand breaks are the most effective way to initiate apoptosis in oocytes of primordial follicles. The widely used catalytic topoisomerase II inhibitor ICRF-187 was able to prevent activation of TAp63α by the topoisomerase II poison doxorubicin but did not prevent oocyte death, suggesting an alternative mechanism of cell death induction.

Patankar, Aniket; Joshi, Kairavi; Sudhakar, Digumarthi V S; Gajbhiye, Rahul; Surve, Suchitra; Parte, Priyanka

doi: 10.1530/rep-24-0382pmid: 40146916

In brief: The transition from histones to protamines during spermiogenesis is critical for male genome integrity and influences fertilisation and early embryogenesis. This study reveals that specific 5′ UTR single nucleotide polymorphisms (SNPs) and promoter methylation of the H2BC1 gene may regulate TSH2B expression in sperm, contributing new insights into the regulatory mechanisms of testis-specific genes and their implications for male fertility.Abstract: The transition from histones to protamines during spermiogenesis plays a crucial role in shaping the male epigenome, and any changes in this process can impact fertilisation potential and the ability of sperm to support early embryogenesis. In our previous research, we observed reduced levels of TSH2B in the sperm of infertile men with oligozoospermia and oligoasthenozoospermia. However, the regulatory mechanisms of the H2BC1 gene, which encodes TSH2B, in the testes are not yet understood. In this study, we investigated whether H2BC1 expression is influenced by SNPs in the 5′ untranslated region (5′ UTR) and promoter methylation. Luciferase assays were performed to assess the impact of 5′ UTR variants in vitro and pyrosequencing was done to evaluate promoter methylation of the H2BC1 gene in the sperm of fertile and infertile men. Our findings suggest that the 5′ UTR variants rs4711096 (c.-83A>G) and rs4712959 (c.-80C>T) positively regulate H2BC1 expression. Methylation analysis indicates hypermethylation of CpG sites, particularly at CpGs 2, 3 and 9 in H2BC1, can influence H2BC1 expression. This study offers new insights into the regulation of testis-specific genes.

Vijay, Sivasatyan; Ibrahim, Siti Fatimah; Osman, Khairul; Zulkefli, Aini Farzana; Mat Ros, Mohd Farisyam; Jamaludin, Norazurashima; Syed Taha, Syed Muhamad Asyraf; Hairulazam, Atikah; Jaffar, Farah Hanan Fathihah

doi: 10.1530/rep-25-0048pmid: 40192818

In brief: Numerous studies have examined the impact of 2.45 GHz Wi-Fi exposure on the testes, highlighting concerns about its potential effects on male fertility. This study extends the investigation beyond the testes to include the epididymis, seminal vesicle and coagulating gland, providing a more comprehensive understanding of how 2.45 GHz Wi-Fi influences the male reproductive system.Abstract: Numerous studies have documented the effect of 2.45 GHz Wi-Fi exposure on the testes and sperm quality. Nevertheless, detailed histological alterations of other male reproductive organs are underexplored. Therefore, this study aimed to evaluate detailed histological alterations of the testes, epididymis, seminal vesicle, coagulating organ and sperm parameters following 2.45 GHz Wi-Fi exposure. Eighteen adult male Sprague Dawley rats (n = 18) were equally divided into three groups (n = 6): control, 4 h and 24 h groups. The groups were exposed to an active router daily for 4 or 24 h, respectively. The control group was sham-exposed using an inactive router. The exposure lasted for 8 weeks at a 20 cm distance, with a power density of 0.141 W/m2 and a specific absorption rate of 0.41 W/Kg. Histological findings revealed vacuolation in the testes and the corpus epididymis of the 4 and 24 h groups. The seminal vesicle in both exposed groups exhibited multifocal atypical hyperplasia. Besides, the seminiferous tubule diameter decreased gradually in both exposed groups, with a substantial decrease in the 24 h group. The spermatogenesis index in 4 and 24 h groups also reduced significantly. The latter result was reflected in the sperm concentration, where both groups showed a significant reduction compared to the control group. Sperm motility also decreased significantly in the 4 h groups. Interestingly, there was a substantial increase in sperm viability in the 24 h group. These findings indicate that 2.45 GHz Wi-Fi exposure causes changes in the histology and histomorphometry measurement and impairs important sperm parameters. This highlights the consequences following Wi-Fi exposure on male reproductive health.

Ning, Anfeng; Xiao, Nansong; Yu, Xiaoqin; Wang, Hu; Guan, Chunyi; Ma, Xu; Xia, Hong-Fei

doi: 10.1530/rep-24-0285pmid: 40193056

In briefEarly pregnancy loss (EPL) is a common pregnancy problem lacking preventive measures. This study shows that DMOG-preconditioned hUC-MSCs can reduce early embryo loss.AbstractEPL, a common pregnancy complication, yet has few effective preventive measures currently. To investigate whether dimethyloxaloylglycine (DMOG)-preconditioned human umbilical cord mesenchymal stem cells (hUC-MSCs) can prevent EPL, we initially evaluated the effect of DMOG on hUC-MSCs in vitro. Subsequently, the DMOG-preconditioned hUC-MSCs were transplanted into the lipopolysaccharide (LPS)-induced murine abortion model for intervention, following which we conducted phenotypic analysis. It was found that DMOG treatment enhanced the mRNA expression of HIF1A, H19 and GLUT1 in hUC-MSCs and augmented their migration capability (P < 0.01). Co-culture experiments showed that DMOG-treated hUC-MSCs notably reduced the mRNA levels of IL6, IL1B and TNFA in LPS-induced HTR-8/SVneo cells (P < 0.01). Moreover, DMOG-preconditioned hUC-MSCs remarkably decreased the fetal resorption rate and increased the embryo weight in LPS-induced abortive mice (P < 0.01). Histological analysis indicated that DMOG-preconditioned hUC-MSCs more effectively promoted their homing and inhibited LPS-induced fibrosis at the maternal–fetal interface. Apart from suppressing inflammatory factors in the serum of pregnant mice, DMOG-preconditioned hUC-MSCs can downregulate the mRNA levels of Il2, Il1b, Tnfa, Il5 and Il9 (P < 0.01), which are pro-inflammatory cytokines secreted by M1 macrophages; and simultaneously upregulate the expression of Cd206 and Pparg (P < 0.01), which serve as the cell surface and nuclear receptors of M2 macrophages in the embryos. Immunofluorescence further verified that the transplantation of DMOG-preconditioned hUC-MSCs could increase the expression of CD206 in embryos. Therefore, DMOG-preconditioned hUC-MSCs might prevent EPL by promoting the transformation of M1 into M2 macrophages.

Cameron, Reese S; Perono, Genevieve A; Natale, Christian D; Petrik, James J; Holloway, Alison C; Hardy, Daniel B

doi: 10.1530/rep-24-0369pmid: 40111139

In brief: This review article summarizes the effects of pre- or peri-conceptual exposure to cannabinoids on female and male reproductive function, along with pregnancy outcomes from 2014 to 2024. In particular, it addresses the gaps in knowledge regarding the specific contributions of the major constituents of cannabis, THC and CBD, on reproduction.Abstract: With increased use of cannabis worldwide, especially in our young adult population, there is a great impetus to understand the impact of cannabis and its constituents (i.e. THC and CBD) on pregnancy, fetal outcomes and male and female reproductive function. This review assessed the current evidence (2014–2024) regarding the effects of cannabinoids on reproductive function (male, female and pregnancy) in animal and human studies. In particular, pre- or periconceptual exposure to cannabinoids were assessed to determine their effects across the lifespan along with transgenerational effects. From the outcomes of this review, we conclude there is a greater need for future preclinical and clinical studies to assess how various routes of cannabinoid exposure along with differing mixtures of cannabinoid constituents may interact to impede reproductive health. Collectively, the outcomes of these studies are important to clinicians and regulatory agencies in the context of functional evidence to support policy and decision-making regarding the safety of cannabis use.

Ning, Anfeng; Xiao, Nansong; Yu, Xiaoqin; Wang, Hu; Guan, Chunyi; Ma, Xu; Xia, Hong-Fei

doi: 10.1530/rep-24-0285pmid: 40193056

In brief: Early pregnancy loss (EPL) is a common pregnancy problem lacking preventive measures. This study shows that DMOG-preconditioned hUC-MSCs can reduce early embryo loss.Abstract: EPL, a common pregnancy complication, yet has few effective preventive measures currently. To investigate whether dimethyloxaloylglycine (DMOG)-preconditioned human umbilical cord mesenchymal stem cells (hUC-MSCs) can prevent EPL, we initially evaluated the effect of DMOG on hUC-MSCs in vitro. Subsequently, the DMOG-preconditioned hUC-MSCs were transplanted into the lipopolysaccharide (LPS)-induced murine abortion model for intervention, following which we conducted phenotypic analysis. It was found that DMOG treatment enhanced the mRNA expression of HIF1A, H19 and GLUT1 in hUC-MSCs and augmented their migration capability (P < 0.01). Co-culture experiments showed that DMOG-treated hUC-MSCs notably reduced the mRNA levels of IL6, IL1B and TNFA in LPS-induced HTR-8/SVneo cells (P < 0.01). Moreover, DMOG-preconditioned hUC-MSCs remarkably decreased the fetal resorption rate and increased the embryo weight in LPS-induced abortive mice (P < 0.01). Histological analysis indicated that DMOG-preconditioned hUC-MSCs more effectively promoted their homing and inhibited LPS-induced fibrosis at the maternal–fetal interface. Apart from suppressing inflammatory factors in the serum of pregnant mice, DMOG-preconditioned hUC-MSCs can downregulate the mRNA levels of Il2, Il1b, Tnfa, Il5 and Il9 (P < 0.01), which are pro-inflammatory cytokines secreted by M1 macrophages; and simultaneously upregulate the expression of Cd206 and Pparg (P < 0.01), which serve as the cell surface and nuclear receptors of M2 macrophages in the embryos. Immunofluorescence further verified that the transplantation of DMOG-preconditioned hUC-MSCs could increase the expression of CD206 in embryos. Therefore, DMOG-preconditioned hUC-MSCs might prevent EPL by promoting the transformation of M1 into M2 macrophages.

Hashim, Prianka H; Perry, Madeline J; Pritchard, Michele T; Gerton, Jennifer L; Duncan, Francesca E

doi: 10.1530/rep-25-0042pmid: 40111977

We synthesize current evidence that granulosa cells possess unique innate immune signaling capabilities. We suggest the novel concept that this serves as a quality control surveillance mechanism by integrating signals from the oocyte and ovarian microenvironment to prevent poor-quality follicles from producing gametes that contribute to the next generation.

Wei, Chenlu; Zeng, Xinxin; Wang, Keer; Wang, Mengchen; Lei, Min; Zhu, Zhenye; Xu, Yining; Zhao, Yanqing; Yang, Qingling; Sun, Yingpu

doi: 10.1530/rep-24-0350pmid: 40110863

In brief: NAD+ levels were reduced in streptozotocin (STZ)-induced diabetic mice, but nicotinamide riboside (NR) supplementation improved these levels in diabetic ovaries and oocytes, enhancing oocyte quality and early embryo development by improving mitochondrial function and lowering reactive oxygen species (ROS) levels.Abstract: Diabetes mellitus is strongly correlated with a decline in oocyte quality; however, noninvasive and effective methods to improve this issue have yet to be fully development. Here, we demonstrate that in vivo supplementation with NR 400 mg/kg/day for 14 days effectively enhances the quality of oocytes from diabetic mice induced by streptozocin 190 mg/kg by restoring nicotinamide adenine dinucleotide (NAD+) levels. NR supplementation not only improved superovulation function of diabetic mice but also improved their oocyte quality and embryonic development potential after fertilization by maintaining normal spindle structure and alleviating mitochondrial dysfunction. In addition, NR supplementation reduced ROS levels in oocytes from diabetic mice. Overall, our findings suggest that dietary NR supplementation is a viable strategy to protect oocytes from diabetes-related deterioration, thereby enhancing reproductive outcomes in maternal diabetes and improving the efficacy of assisted reproductive technology.

Abad, Cilia; Musilova, Ivana; Cifkova, Eva; Portillo, Ramon; Kumnova, Fiona; Karahoda, Rona; Sterba, Martin; Lisa, Miroslav; Kacerovsky, Marian; Stranik, Jaroslav; Stuchlik, Ales; Staud, Frantisek

doi: 10.1530/rep-24-0378pmid: 40192828

In brief: Intrauterine inflammation disrupts tryptophan metabolism in both the placenta and the fetal brain, leading to a shift toward neurotoxic metabolites. These findings highlight the critical role of placental function in neurodevelopment and suggest that inflammation-induced metabolic changes may contribute to neurodevelopmental disorders.Abstract: The placenta plays a crucial role beyond nutrient transfer, acting as a dynamic endocrine organ that significantly influences maternal physiology and fetal development. It responds rapidly to even slight changes in the in utero environment to promote fetal survival. Disruptions in placental function are increasingly recognized as key contributors to the origins of neurodevelopmental disorders. In this study, we employed advanced technology to induce intrauterine inflammation through ultrasound-guided administration of LPS into gestational sacs. We then evaluated its effects on the gene expression of enzymes involved in TRP metabolism and conducted a comprehensive LC/MS analysis of the metabolome in the placenta and fetal brain of Wistar rats. Our results show that intraamniotic injection of LPS induces a robust inflammatory response leading to significant alterations in TRP metabolism, including downregulation of tryptophan hydroxylase (TPH) in the placenta, resulting in a decrease in serotonin (5-HT) levels. Similarly, in the fetal brain, exposure to LPS led to reduced Tph expression and increased monoamine oxidase expression, suggesting a decrease in 5-HT synthesis and an increase in its degradation. Furthermore, an upregulation of the kynurenine pathway was observed in both the placenta and fetal brain. Moreover, we detected a shift toward neurotoxicity, evidenced by an imbalance between neuroprotective and neurotoxic metabolites, including decreased levels of kynurenic acid and upregulation of kynurenine monooxygenase in the fetal brain. In conclusion, our findings reveal significant alterations in TRP metabolism following intrauterine inflammation, potentially contributing to neurodevelopmental disorders.