{"title":"Impact of culture media pre-equilibration methods on embryo development","authors":"Dapeng Chu, Yao Fu","doi":"10.1016/j.repbio.2024.100897","DOIUrl":"10.1016/j.repbio.2024.100897","url":null,"abstract":"<div><p>This study investigates the influence of four culture media pre-equilibration methods on embryo development and clinical pregnancy outcomes. The methods are as follows: Method A involved covering media with fresh mineral oil in humid-type incubators for 24 h. Method B replicated Method A in dry-type incubators. Method C utilized pre-equilibrated (humidified) mineral oil to cover the media, also in humid-type incubators for 24 h. Method D followed the same process as Method C but in dry-type incubators. Subsequently, media from all groups were transferred to dry-type incubators for 72 h. Osmolality was measured at 24, 48, 72, and 96 h. For G1 PLUS, no significant differences were observed among groups at 24, 48, and 72 h. However, at 96 h, Groups B and D exhibited significantly higher osmolality than Groups A and C (A vs B, <em>p</em> = 0.043; A vs D: <em>p</em> = 0.046; B vs C, <em>p</em> = 0.043; C vs D, <em>p</em> = 0.046). No significant variations were found between Groups A and C or B and D. Similar results were obtained for G2 PLUS. A retrospective analysis of embryo development and clinical outcomes using Methods A revealed significant improvements in good blastocysts and available embryos compared with Method B for all (<em>p</em> = 0.005 and 0.004) and IVF cycles (<em>p</em> = 0.025 and 0.017). Method A also enhanced blastocyst formation in ICSI cycles (<em>p</em> = 0.017). However, clinical pregnancy outcomes did not significantly differ between Methods A and B. Pre-equilibrating culture media overnight in humid-type incubators, even when covered with fresh mineral oil, significantly mitigates osmolality rise and improves embryo development potential during embryo culture in dry-type incubators.</p></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"24 3","pages":"Article 100897"},"PeriodicalIF":2.1,"publicationDate":"2024-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141176988","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Kamila Gorczyca , Małgorzata M. Kozioł , Żaneta Kimber-Trojnar , Joanna Kępa , Małgorzata Satora , Anna K. Rekowska , Bożena Leszczyńska-Gorzelak
{"title":"Premature rupture of membranes and changes in the vaginal microbiome – Probiotics","authors":"Kamila Gorczyca , Małgorzata M. Kozioł , Żaneta Kimber-Trojnar , Joanna Kępa , Małgorzata Satora , Anna K. Rekowska , Bożena Leszczyńska-Gorzelak","doi":"10.1016/j.repbio.2024.100899","DOIUrl":"https://doi.org/10.1016/j.repbio.2024.100899","url":null,"abstract":"<div><p>Preterm birth affects approximately 15 million women worldwide, of which 30 % is due to preterm premature rupture of membranes (PPROM). The reasons for shortening the duration of pregnancy are seen in genetic, hormonal, immunological and socio-economic conditions. Recent years have provided a lot of evidence on the impact of the microbiota and whole microbiome on pregnant women, suggesting that the microorganisms inhabiting the vagina significantly affect the risk of preterm delivery. The aim of the study was to review studies evaluating the composition of the vaginal microflora and its role in the occurrence of preterm labor caused by PPROM, and to evaluate the potential beneficial effect of probiotics on preventing the development of preterm labor. Vaginal microbial dysbiosis is observed in PPROM, which, due to its association with a high risk of prematurity and infection, increases neonatal morbidity and mortality. Further research on biomarkers for screening, early prognosis and diagnosis of PPROM seems advisable. Probiotics as a potential intervention can prevent the development of pathological vaginal flora, reducing the risk of infection in women planning pregnancy and pregnant women.</p></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"24 3","pages":"Article 100899"},"PeriodicalIF":2.1,"publicationDate":"2024-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141156401","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Claudia Omes , Alice Conti , Laura Benedetti , Veronica Tomasoni , Davide De Marchi , Rossella E. Nappi , M. Gabriella Cusella De Angelis , Gabriele Ceccarelli
{"title":"Expression of miRNA from spent pre-implantation embryos culture media","authors":"Claudia Omes , Alice Conti , Laura Benedetti , Veronica Tomasoni , Davide De Marchi , Rossella E. Nappi , M. Gabriella Cusella De Angelis , Gabriele Ceccarelli","doi":"10.1016/j.repbio.2023.100847","DOIUrl":"https://doi.org/10.1016/j.repbio.2023.100847","url":null,"abstract":"<div><p>This study examines the expression of three microRNAs (hsa-miR-661, hsa-miR-21–5p, hsa-miR-372–5p) in spent pre-implantation embryos culture media to identify possible new non-invasive biomarkers of embryo competence, predictive of development to the blastocyst stage. A preliminary analysis on 16 patients undergoing IVF cycles was performed by collecting and stored spent culture media on the fifth/sixth day of embryo culture. Expression of miRNAs was evaluated according to the embryos’ fate: 1) NE/DG: non-evolved or degenerate embryos; 2) BLOK: embryos developed to the blastocyst stage. Preliminary results revealed a higher miRNAs expression in NE/DG spent media. To elucidate the roles of these miRNAs, we employed a robust bioinformatics pipeline involving: 1) in-silico miRNA Target Prediction using RNAHybrid, which identified the most-likely gene targets; 2) Construction of a Protein-Protein Interaction network via GeneMania, linking genes with significant biological correlations; 3) application of modularity-based clustering with the gLay app in Cytoscape, resulting in three size-adapted subnets for focused analysis; 4) Enrichment Analysis to discern the biological pathways influenced by the miRNAs. Our bioinformatics analysis revealed that hsa-miR-661 was closely associated with pathways regulating cell shape and morphogenesis of the epithelial sheet. These data suggest the potential use of certain miRNAs to identify embryos with a higher likelihood of developing to the blastocyst stage. Further analysis will be necessary to explore the reproducibility of these findings and to understand if miRNAs here investigated can be used as biomarkers for embryo selection before implantation into the uterus or if they may be reliable predictors of IVF outcome.</p></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"24 2","pages":"Article 100847"},"PeriodicalIF":2.1,"publicationDate":"2024-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141078549","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yunqing Li , Yulan Zhou , Tianzhong Ma , Jiaze Dai , Hongbo Li , Qingjun Pan , Wenying Luo
{"title":"Research progress on the role of autophagy in the development of varicocele","authors":"Yunqing Li , Yulan Zhou , Tianzhong Ma , Jiaze Dai , Hongbo Li , Qingjun Pan , Wenying Luo","doi":"10.1016/j.repbio.2024.100894","DOIUrl":"https://doi.org/10.1016/j.repbio.2024.100894","url":null,"abstract":"<div><p>Varicocele (VC) is a common cause of infertility in men. Pathophysiological changes caused by VC, such as testicular hypoxia, high temperatures, oxidative stress, abnormal reproductive hormones, and Cd accumulation, can induce autophagy, thus affecting the reproductive function in patients with this condition. Autophagy regulators can be classified as activators or inhibitors. Autophagy activators upregulate autophagy, reduce the damage to the testis and epididymis, inhibit spermatogenic cell apoptosis, and protect fertility. In contrast, autophagy inhibitors block autophagy and aggravate the damage to the reproductive functions. Therefore, elucidating the role of autophagy in the occurrence, development, and regulation of VC may provide additional therapeutic options for men with infertility and VC. In this review, we briefly describe the progress made in autophagy research in the context of VC.</p></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"24 2","pages":"Article 100894"},"PeriodicalIF":2.1,"publicationDate":"2024-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141078514","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Effect of low ethanol concentration in maturation medium on developmental ability, mitochondria, and gene expression profile in mouse oocytes","authors":"Susaki Kizasu , Takuya Sato , Yuki Inoue , Hidetaka Tasaki , Komei Shirasuna , Yuichi Okiishi , Hisataka Iwata","doi":"10.1016/j.repbio.2023.100854","DOIUrl":"https://doi.org/10.1016/j.repbio.2023.100854","url":null,"abstract":"<div><p>Ethanol affects pre-conceptional oocyte quality in women. In this study, we examined the effect of low ethanol concentrations on mouse oocytes. Oocytes were collected from the ovaries of 9–10 week old mice and allowed to mature in vitro in the presence of low concentrations of ethanol (0.1% and 0.2% v/v) for 24 h. Treatment of oocytes with ethanol (0.2%) during maturation decreased the mitochondrial DNA content and membrane potential compared to that in untreated ones, whereas the ATP content did not differ between the groups. Both 0.1% and 0.2% ethanol reduced the lipid content in the oocytes. In addition, immunostaining revealed that oocytes cultured in maturation medium containing ethanol (0.2%) had reduced levels of global DNA methylation and DNMT3A compared with untreated oocytes, and decreased rate of blastocyst development with low mitochondrial protein levels (TOMM40) in embryo. RNA-sequencing of the ethanol-treated (0.2%) and untreated oocytes revealed that mitochondria were a major target of ethanol. In conclusion, treatment of oocytes with low concentration of ethanol reduces the developmental rate to the blastocyst stage, with a lower total cell number and global DNA methylation. In addition, ethanol affected mitochondrial function and mitochondria-related gene expression.</p></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"24 2","pages":"Article 100854"},"PeriodicalIF":2.1,"publicationDate":"2024-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141073365","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Guohui Zhang , Dongsheng Xiong , Fei Ye , Yuhong Zhao , Xinrong Du , Weiwei Zhi , Fulin Liu , Jiuzhi Zeng , Wenming Xu , Weixin Liu , Yi Shi
{"title":"A Key regulatory protein QRICH2 governing sperm function with profound antioxidant properties, enhancing sperm viability","authors":"Guohui Zhang , Dongsheng Xiong , Fei Ye , Yuhong Zhao , Xinrong Du , Weiwei Zhi , Fulin Liu , Jiuzhi Zeng , Wenming Xu , Weixin Liu , Yi Shi","doi":"10.1016/j.repbio.2024.100881","DOIUrl":"https://doi.org/10.1016/j.repbio.2024.100881","url":null,"abstract":"<div><p>Infertility poses a global health and social challenge, affecting approximately 15% of couples at childbearing age, with half of the cases attributed to male factors, wherein genetic factors exert a substantial role. In our prior investigation, we identified loss-of-function variants within the gene encoding glutamine-rich protein 2 (QRICH2) in two consanguineous families, leading to various morphological abnormalities in sperm flagella and male infertility. Moreover, our observations in <em>Qrich2</em> knockout mice revealed a pronounced reduction in spermatozoa count. However, the underlying mechanism remains elusive, prompting further investigation in the current study. By conducting experiments such as Hematoxylin-eosin (HE) staining, immunofluorescence staining, flow cytometry, and single sperm metabolism analysis on the testes and spermatozoa of <em>Qrich2</em> knockout mice, we found a strong antioxidant capacity mediated by QRICH2 both <em>in vivo</em> and <em>in vitro</em>. <em>Qrich2</em> knockout led to elevated levels of ROS, consequently inducing DNA damage in spermatids, which in turn triggered increased autophagy and apoptosis, ultimately causing a significant decrease in spermatozoa count. Incubation with the N-terminal purified protein of QRICH2 exhibited potent strong antioxidant activity at the cell and spermatozoa levels <em>in vitro</em>, thereby enhancing spermatozoa viability and motility. Therefore, QRICH2 plays a crucial role in safeguarding spermatids from excessive ROS-induced damage by augmenting antioxidant capacity, thereby promoting spermatozoa survival and improving motility. Furthermore, the N-terminal purified protein of QRICH2 shows promise as an additive for protecting spermatozoa during preservation and cryopreservation.</p></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"24 2","pages":"Article 100881"},"PeriodicalIF":2.1,"publicationDate":"2024-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141072931","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Zuzana Fabová , Barbora Loncová , Abdel Halim Harrath , Alexander V. Sirotkin
{"title":"Involvement of circular RNAs in the control of porcine ovarian cell functions: Upregulation by ciR-00596 and downregulation by ciR-00646","authors":"Zuzana Fabová , Barbora Loncová , Abdel Halim Harrath , Alexander V. Sirotkin","doi":"10.1016/j.repbio.2024.100860","DOIUrl":"10.1016/j.repbio.2024.100860","url":null,"abstract":"<div><p>The current understanding of the role of circular RNAs (circRNAs) in regulating ovarian functions is inadequate. To assess the impact of ciR-00596 and ciR-00646 on the regulation of basic porcine ovarian granulosa cell functions, we conducted upregulation (utilizing overexpressing vectors) and downregulation (utilizing shRNA vectors) of these circRNAs. The relative expression of both circRNAs, cell viability and proliferation (accumulation of PCNA, cyclin B1, and XTT-positive cells), cytoplasmic (accumulation of bax and caspase-3) and nuclear (DNA fragmentation) apoptosis, and the release of progesterone, testosterone, estradiol, IGF-I, and oxytocin were evaluated. Transfection of cells with the ciR-00596 overexpression vector resulted in increases in cell viability and proliferation and the release of progesterone and IGF-I, while it decreased the cytoplasmic and nuclear apoptosis, testosterone, estradiol, and oxytocin output. CiR-00596 inhibition had the opposite effects. The overexpression of ciR-00646 decreased cell viability and proliferation, and the release of progesterone, IGF-I, and oxytocin, while increasing cytoplasmic and nuclear apoptosis and the output of testosterone and estradiol. Our findings are the first to show the stimulatory action of ciR-00596 and the inhibitory effect of ciR-00646 on ovarian cell functions, including the cell cycle, apoptosis, and secretory activity.</p></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"24 2","pages":"Article 100860"},"PeriodicalIF":2.1,"publicationDate":"2024-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141066333","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"LncRNA HOTAIR promotes the migration and invasion of cervical cancer through DNMT3B/LATS1/ YAP1 pS127 axis","authors":"Zhihao Zhang , Xianyi Zhou , Jiulin Li , Qinghui Meng , Peng Zheng","doi":"10.1016/j.repbio.2024.100893","DOIUrl":"https://doi.org/10.1016/j.repbio.2024.100893","url":null,"abstract":"<div><p>Metastasis is the hallmark of cancer that is responsible for the greatest number of cancer-related deaths. As a critical regulator of the Hippo pathway, the phosphorylation status of Yes-associated protein 1 (YAP1), mainly at S127, is critical for its oncogenic function. Herein, we aim to investigate the precise molecular mechanism between long noncoding RNA HOX transcript antisense RNA (HOTAIR) and YAP1 phosphorylation in regulating tumor migration and invasion. In this study, we showed that inhibition of HOTAIR significantly decreased the migration and invasion of cancer cells both in vitro and in vivo through elevating the phosphorylation level of YAP1 on serine 127, demonstrating a tumor suppressive role of YAP1 S127 phosphorylation. Through bisulfite sequencing PCR (BSP), we found that inhibition of HOTAIR dramatically increased Large Tumor Suppressor Kinase 1 (LATS1) expression by regulating LATS1 methylation via DNA methyltransferase 3β (DNMT3B). In accordance with this observation, DNMT3B just only altered the distribution of YAP1 in the cytoplasm and the nucleus by inhibiting its phosphorylation, but did not change its total expression. Mechanistically, we discovered that HOTAIR suppressed YAP1 S127 phosphorylation by regulating the methylation of LATS1 via DNMT3B, the consequence of which is the translocation of YAP1 into the nucleus, reinforcing its coactivating transcriptional function, which in turn promotes the migration and invasion of cancer cells. Collectively, our data reveal that the phosphorylation of YAP1 S127 plays a vital role in the function of HOTAIR in tumorigenicity, and should be taken into consideration in future therapeutic strategies for cervical cancer.</p></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"24 2","pages":"Article 100893"},"PeriodicalIF":2.1,"publicationDate":"2024-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140950433","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Andrea Fernández-Montoro , Daniel Angel-Velez , Berta Cava-Cami , Osvaldo Bogado Pascottini , Krishna Chaitanya Pavani , Katrien Smits , Ann Van Soom
{"title":"How to beat the bull: Lycopene as a tool to improve in vitro fertilization efficiency in bulls with high polyspermy","authors":"Andrea Fernández-Montoro , Daniel Angel-Velez , Berta Cava-Cami , Osvaldo Bogado Pascottini , Krishna Chaitanya Pavani , Katrien Smits , Ann Van Soom","doi":"10.1016/j.repbio.2024.100888","DOIUrl":"10.1016/j.repbio.2024.100888","url":null,"abstract":"<div><p>High levels of reactive oxygen species (ROS) derived from in vitro conditions compromise oocyte quality and subsequent polyspermy prevention by the zona and membrane block. Antioxidant supplementation, like lycopene, during in vitro maturation (IVM) mitigates ROS effects, yet, its efficacy in blocking polyspermy remains uncertain. This study aims to evaluate the effect of lycopene supplementation during IVM on oocyte maturation, fertilization, and developmental parameters. To this end, bovine oocytes were supplemented with 0.2 µM lycopene and fertilized with semen from three bulls. The three bulls showed different fertilization potential in vitro, with bull 1 showing the highest penetration and polyspermy rates and the lowest in vitro fertilization (IVF) efficiency. Interestingly, in bull 1, the treatment with lycopene improved IVF efficiency (p = 0.043) and reduced the polyspermy rate (p = 0.028). However, none of these effects were observed in bulls 2 and 3. Bulls with higher penetration rates exhibited better blastocyst rates although those rates did not seem to be associated with polyspermy or IVF efficiency. Oocyte mitochondrial distribution and activity and cortical granule migration and distribution were not influenced by lycopene. In conclusion, we demonstrated that lycopene addition during oocyte maturation had a positive impact on IVF efficiency by reducing polyspermy rates in a bull-dependent manner. The reduction in polyspermy rates was not caused by changes in cortical granule migration or oocyte mitochondrial distribution. Lycopene must therefore induce other changes in the oocyte that lower the in vitro penetration rates of specific bulls prone to polyspermy.</p></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"24 2","pages":"Article 100888"},"PeriodicalIF":2.1,"publicationDate":"2024-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140946747","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}