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Chinelo Uju C Uju, Department of Veterinary Biomedical Sciences, University of Saskatchewan, Saskatoon, Canada

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Katayoon Karimzadeh K Karimzadeh, Department of Veterinary Biomedical Sciences, University of Saskatchewan, Saskatoon, Canada

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Suraj Unniappan S Unniappan, Department of Veterinary Biomedical Sciences, University of Saskatchewan, Saskatoon, S7N 5A2, Canada

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Brain-derived neurotrophic factor (BDNF) is a peptide widely known for its role in neurogenesis and synaptic plasticity. Its expression in non-neuronal tissues has been reported. In mammals, it is involved in ovarian development, follicle growth, oocyte maturation, and early embryonic development. In zebrafish, it was demonstrated that BDNF increases food intake and regulates metabolism. Reproduction and metabolism are tightly linked. We hypothesized that BDNF modulates reproductive hormones and reproductive functions in zebrafish. This study aimed to determine BDNF expression in the zebrafish reproductive axis and whether it modulates the reproductive endocrine milieu and oocyte biology in zebrafish. Our results show that bdnf and its receptor trkb, and BDNF-like immunoreactivity are detected in zebrafish gonads and liver cells. This suggests BDNF local production and possible actions within the gonads and liver. Intraperitoneal administration of 1, 10, or 100 ng/g bodyweight BDNF significantly (ANOVA, p<0.05) increased sgnrh/cgnrh-II, kiss1, and cyp19a1b mRNAs in the zebrafish brain; steroidogenic enzymes (star and cyp19a1a) and key receptors in the zebrafish gonads. In vitro incubation of zebrafish liver cells with BDNF significantly (ANOVA, p<0.05) increased estrogen receptor mRNAs and vitellogenin concentrations (ELISA) in the cells. BDNF (100 ng/mL) induced (ANOVA, p<0.05) oocyte maturation in vitro at 24 hours post-incubation and significantly upregulated cumulus-expansion related genes (ANOVA, p<0.05). Overall, our findings indicate a stimulatory role for BDNF in the reproductive axis of zebrafish. This provides impetus for future research on its mechanism of action and potential practical applications to enhance reproduction in aquaculture.

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Nanami Sono Graduate School of Environmental Science and Technology, Okayama University, Tsushimanaka, Kita, Okayama, Japan

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Mone Takeshita Graduate School of Environmental Science and Technology, Okayama University, Tsushimanaka, Kita, Okayama, Japan

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Mizuho Chikushi Graduate School of Environmental Science and Technology, Okayama University, Tsushimanaka, Kita, Okayama, Japan

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Saki Nakashima Faculty of Agriculture, Okayama University, Tsushimanaka, Kita, Okayama, Japan

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Shoko Miyawaki Faculty of Agriculture, Okayama University, Tsushimanaka, Kita, Okayama, Japan

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Misaki Wakamatsu Faculty of Agriculture, Okayama University, Tsushimanaka, Kita, Okayama, Japan

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Yasuhiro Fujiwara Laboratory of Pathology and Development, Institute for Quantitative Biosciences, The University of Tokyo, Yayoi, Bunkyo, Tokyo, Japan

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Tetsuo Kunieda Faculty of Veterinary Medicine, Okayama University of Science, Imabari, Ehime, Japan

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Junko Otsuki Graduate School of Environmental Science and Technology, Okayama University, Tsushimanaka, Kita, Okayama, Japan
Assisted Reproductive Technology Center, Okayama University, Tsushimanaka, Kita, Okayama, Japan

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In brief

Repro57 mice, bearing an Rnf212 gene mutation, exhibit infertility in both homozygous mutant males and females, revealing arrested spermatogenesis in males and investigating unclear mechanisms in females. The study highlights aneuploidy and altered kinetochore patterns in repro57 homozygous mutant oocytes, which impact later stages of embryo development.

Abstract

Repro57 mice, induced with N-ethyl-N-nitrosourea and harboring a mutation in the Rnf212 gene, exhibit infertility in both homozygous mutant males and females. Rnf212 plays a crucial role in recombination and crossover designation. In male repro57 homozygous mutants, spermatocytes often degenerate during late prophase, and mature spermatozoa are absent in the seminiferous epithelium, indicating arrested spermatogenesis as the cause of infertility. Despite reports of infertility in Rnf212-knockout female mice, the specific mechanisms underlying infertility in female repro57 homozygous mutants remain elusive. This study investigates the chromosomal and kinetochore patterns of mature oocytes and their developmental potential following in vitro fertilization in female repro57 homozygous mutant mice. While all wild-type oocytes progress to metaphase II and exhibit euploidy, all repro57 homozygous mutant mouse oocytes display aneuploidy. Additionally, kinetochore distances in repro57 homozygous mutant oocytes exceed those observed in wild-type counterparts. Although no significant differences are noted in fertilization and early embryo development rates between wild-type and repro57 homozygous mutant mice, embryos derived from repro57 homozygous mutants exhibit significantly lower morula and blastocyst rates, accompanied by frequent cytokinesis failure and vacuole formation. These findings suggest that the premature segregation of sister chromatids in repro57 homozygous mutant mice adversely impacts the later stages of embryo development.

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Julie Gardella J Gardella, Environmental Medicine, NYU Langone Health, New York, United States

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Dimitri Abrahamsson D Abrahamsson, Environmental Pediatrics, NYU Langone Health Department of Pediatrics, New York, United States

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Judith Zelikoff J Zelikoff, Environmental Medicine, NYU Langone Health, New York, United States

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Those undergoing pregnancy are often excluded from clinical drug trials due to the risk that participation would pose. However, they often require pharmaceuticals to manage health conditions that, if gone untreated, could harm themselves or the fetus. This can mean that such individuals take one or more pharmaceuticals during pregnancy, many of which have unknown reproductive effects. Machine learning models have been used to successfully predict a number of reproductive toxicological outcomes for pharmaceuticals, including transplacental transfer, US Food and Drug Administration safety rating, and drug interactions. Models use quantitative chemical and structural features of active compounds to make predictions concerning the outcome of interest using computational algorithms. Results from these models can be a potential source of valuable information for pregnant people and their medical providers when making decisions regarding therapeutic drug use. This review summarizes current machine learning applications to make predictions about risk and toxicity of medication use during pregnancy. Our review of the recent literature revealed that machine learning quantitative structure-activity relationship models can be used successfully to predict the transplacental transfer and the US Food and Drug Administration pregnancy safety category of pharmaceuticals; such models have also been employed to predict drug interactions, though not specifically during pregnancy. This latter topic is a potential area for future research. In this review, no single algorithm or descriptor-calculation software emerged as the most widely used, and their performances depend on a variety of factors, including outcome of interest and combination of such algorithms and software.

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Kirsten Das K Das, NICHD, National Institutes of Health, Bethesda, United States

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Veronica Gomez-Lobo V Gomez-Lobo, NICHD, National Institutes of Health, Bethesda, United States

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Contraception care is an important aspect of comprehensive gender affirming care in transmasculine and gender non-binary (TGD) adolescent and young adults (AYA). TGD AYA seek contraception care for a variety of reasons and there are several important considerations that are unique to this population. In addition to pregnancy prevention, transmasculine and gender nonbinary persons may seek contraception to achieve amenorrhea, reduce gender dysphoria, or as part of their gender affirming journey. While patients on testosterone therapy often experience amenorrhea, pregnancy has been reported and testosterone is not a form of contraception. Visits for contraceptive counseling provide an opportunity to discuss fertility goals and preservation planning as well as the possibility that some methods may provide endometrial protection. In this narrative review, we analyze the current literature and discuss specific considerations for contraceptive use in this population. We also propose a contraception overview for shared decision making for patients and their clinicians.

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Yan Gu Y Gu, Reproductive Medicine Center, The Second Hospital of Tianjin Medical University, Tianjin, China

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Ning Song N Song, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

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Long Yang L Yang, Reproductive Health institute, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Shanghai, China

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Yan Shi Y Shi, NHC Key Lab of Reproduction Regulation, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Shanghai, China

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Hao-Ran Xu H Xu, Medical School, Fudan University, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Shanghai, China

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Ya-Wei Xin Y Xin, Reproductive Medicine Center, The Second Hospital of Tianjin Medical University, Tianjin, China

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Ling-Han Chen L Chen, Hangzhou Yuhang Center for Disease Control and Prevention, Hangzhou Yuhang Center for Disease Control and Prevention, Hangzhou, China

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Wen-Wen Gu W Gu, Medical School, Fudan University, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Shanghai, 200032, China

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Jian Wang J Wang, School of Pharmacy, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Shanghai, China

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Placenta-associated pathologies, including early pregnancy loss (EPL) and preeclampsia (PE), share a common phenomenon of insufficient extravillous trophoblasts (EVTs) invasion. It was previously observed that down-regulated miR-486-5p expression inhibited the invasion of EVTs, and a decreased peripheral miR-486-5p was associated with EPL. However, the exact roles of miR-486-5p played in pathogenesis of EPL, as well as the molecular pathway underlying roles of miR-486-5p in EVTs invasion, remains poorly understood. In this study, a decreased miR-486-5p expression in uterine embryo implantation site at gestation day (GD) 8.5, and an increased expression of Smad2, a target of miR-486-5p, were observed in the lipopolysaccharide (LPS)-induced EPL mouse model. The invasion and viability of immortalized human EVTs line, HTR-8/SVneo, were inhibited by LPS, accompanied with a reduced miR-486-5p expression. LPS showed a promoting effect on the Smad2 expression, of which could be attenuated by miR-486-5p mimics. And the down-regulated Smad2 could effectively restore the impaired invasion and viability of HTR-8/SVneo cells caused by LPS or miR-486-5p inhibitor. Furthermore, LPS could promote the TNFα production in HTR-8/SVneo cells, whereas both of siSmad and miR-486-5p mimics could reverse such an effect. By analyzing the human decidua single-cell RNA sequencing and transcriptome datasets derived from the Gene Expression Omnibus, it was found that, compared to control early pregnant women, the Smad2 expression was significantly increased in recurrent miscarriages (RM) patients. Collectively, these data suggested that, decreased miR-486-5p expression might lead to EPL at least partially by inhibiting invasion and/or promoting TNFα production of EVTs via targeting Smad2.

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Jiaojiao Huang College of Life Science, Qingdao Agricultural University, Qingdao, China

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Xinyu Li College of Life Science, Qingdao Agricultural University, Qingdao, China

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Dongyu Zhang College of Animal Science and Technology, Qingdao Agricultural University, Qingdao, China

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Luzhen Wang College of Animal Science and Technology, Qingdao Agricultural University, Qingdao, China

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Zhao Li Department of Pharmacology, Qingdao University School of Pharmacy, Qingdao, China

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Zhenhua Song Department of Pharmacology, Qingdao University School of Pharmacy, Qingdao, China

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In brief

Male reproductive problems under psychological stress were widely studied. Using chronically unpredictable mild stress-treated mice, we found that reduced serum testosterone levels were related to the low level of cholesterol in the Leydig cells.

Abstract

Testosterone deficiency in humans can be caused by depressive symptoms; however, the causes of this deficiency are incompletely understood. This study demonstrates that male mice with depression-like symptoms due to chronic unpredictable mild stress (CUMS) show reduced serum testosterone levels and disrupted sexual behaviors. However, the observed testosterone reductions were not caused by apoptosis of Leydig cells. Oil red O staining revealed that lipid droplets were dramatically decreased in Leydig cells, suggesting that defects in cholesterol uptake might be related to testosterone deficiency in depression-like mice. To investigate the potential mechanism, lipid homeostasis was examined by liquid chromatography-tandem mass spectrometry. The results revealed that higher levels of sphingomyelins (SM 8:0;2O/28:1, 18:0;2O/22:2, 33:0;3O, 33:1;2O) were linked to decreased cholesterol levels. Further investigation indicated that testosterone biosynthesis from cholesterol in Leydig cells was impaired by the downregulation of Ldlr, Srb1, Lhr, and P450scc. Elevated levels of interferon signaling-associated pathways in depression-like mice testes may also contribute to decreased testosterone levels. Taken together, these findings provide a novel understanding of male reproductive problems under psychological stress and suggest that cholesterol uptake might be a causal factor in reduced testosterone production in depression-like mice.

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Teppei Takeshima Department of Urology, Reproduction Center, Yokohama City University Medical Center, Yokohama, Kanagawa, Japan

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Jurii Karibe Department of Urology, Reproduction Center, Yokohama City University Medical Center, Yokohama, Kanagawa, Japan

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Shinnosuke Kuroda Department of Urology, Reproduction Center, Yokohama City University Medical Center, Yokohama, Kanagawa, Japan

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Yasushi Yumura Department of Urology, Reproduction Center, Yokohama City University Medical Center, Yokohama, Kanagawa, Japan

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To enhance surgical testicular sperm retrieval outcome for men with nonobstructive azoospermia, a deep-learning model was developed to identify positive seminiferous tubules by labeling 110 images with sperm-containing tubules sampled during microdissection testicular sperm extraction as training and validation data. After training, the model achieved an average precision of 0.60.

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Duygu Yaman Gram Department of Physiology, Faculty of Veterinary Medicine, Erciyes University, Turkey

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Murat Abay Department of Obstetrics and Gynecology, Faculty of Veterinary Medicine, Erciyes University, Turkey

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Narin Liman Department of Histology and Embryology, Faculty of Veterinary Medicine, Erciyes University, Turkey

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Muhittin Tekin Clinical Unit for Herd Health Management in Ruminants, Center for Veterinary Systems Transformation and Sustainability, Clinical Department for Farm Animals and Food System Science, University of Veterinary Medicine Vienna, Austria

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Mariusz P Kowalewski Institute of Veterinary Anatomy, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland

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Aykut Gram Department of Histology and Embryology, Faculty of Veterinary Medicine, Erciyes University, Turkey

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In brief

Activation of TLR2/TLR1 alters in vitro formation of capillary-like structures and induces inflammatory processes in ovine luteal endothelial (OLENDO) cells.

Abstract

Postpartum bacterial infections of the uterus affect uterine physiology and ovarian activity, causing fertility problems. The outer membrane component of Gram-negative bacteria, lipopolysaccharide, is involved in the initiation of the local inflammatory processes, and other bacterial toxins, particularly lipopeptides, have also been shown to be potent cytokine inducers, acting via Toll-like receptor-2 (TLR2). However, the possible adverse effects of TLR2 on ovarian and luteal activities have not yet been investigated in depth. The strong expression of TLR2 in the blood vessels of the corpus luteum led us to hypothesize that TLR2 activation might participate in the disruption of luteal vascular functionality. Therefore, we analyzed the effects of Pam3CSK4 (Pam3CysSerLys4), a synthetic triacylated lipopeptide and TLR2/TLR1 ligand, on the functionality of gap junctional intercellular communication (GJIC), endothelial cell invasion, and in vitro capillary-like network formation in an immortalized ovine luteal endothelial (OLENDO) cell line. Pam3CSK4 treatment of OLENDO cells disrupted in vitro tube formation but had no effect on GJIC or migration of OLENDO cells. Furthermore, Pam3CSK4 induced the expression of NFKB, IL6, and IL8 in OLENDO cells. Additionally, the basal availability of TLRs (TLR1–10) and TLR co-receptors (MYD88, LY96/MD2, and CD14) in OLENDO cells was confirmed by conventional PCR. Finally, the activation of TLR2/TLR1 appears to alter in vitro formation of capillary-like structures and induce inflammatory processes in OLENDO cells.

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Elizabeth Rubin E Rubin, Reproductive Endocrinology and Infertility, Oregon Health and Science University, Portland, United States

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Marissa Palmor M Palmor, Reproductive Endocrinology and Infertility, Oregon Health and Science University, Portland, United States

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Paula Amato P Amato, Reproductive Endocrinology and Infertility, Oregon Health and Science University, Portland, United States

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Years of growing research demonstrate that transgender and gender diverse (TGD) people desire fertility counseling and family building, however social and medical factors can impact future fertility options. Fortunately, TGD individuals have many viable options for family building using their own gametes and/or reproductive organs. However, the nuanced ways in which different gender affirming treatments affect reproduction, the interplay with non-treatment related infertility factors, and mitigation of likely dysphoria triggers are all critical to actual utilization. This review focuses on fertility treatment and preservation options for TGD patients and highlights these influential social and medical factors. Fertility treatments may be associated with worsening gender dysphoria in TGD people, and an affirming clinical environment and conscientious provider approach is paramount to treatment success. However, reducing gender dysphoria can also require specific changes to medically assisted reproduction and sperm collection protocols, some which carry the potential for diminished outcomes or unknown effects. Adolescents undergoing fertility preservation treatments may need more support or additional protocol modifications, and outcomes may be poorer in this age group compared with adults. Testicular and ovarian tissue cryopreservation may present a fertility preservation option for prepubertal TGD children, however in-vitro gamete maturation remains experimental in this situation.

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Isha Singh I Singh, Department of Gamete Immunobiology, ICMR-National Institute for Research in Reproductive and Child Health, Mumbai, India

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Priyanka Parte P Parte, Department of Gamete Immunobiology, ICMR-National Institute for Research in Reproductive and Child Health, Mumbai, India

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The histone variant TH2B, enriched in oocytes, sperm, and early embryos, decreases as embryos differentiate into pre-gastrula stages. Despite its presence, the role of TH2B in epigenetic reprogramming during early embryonic development remains largely under-researched. Our study employed ultra-low-input ChIP-seq (ULI-ChIP) to analyze the genome-wide distribution of TH2B in MII oocytes and early embryos. We found that TH2B is enriched in the chromatin of oocytes and 2-cell stage embryos but becomes less prevalent after the 2-cell stage. Correlation analysis revealed that the TH2B chromatin patterns in sperm and preimplantation embryos are more similar to each other than to those in MII oocytes. Gene ontology (GO) analysis of TH2B-occupied loci linked them to various developmental processes, including oogenesis, fertilization, chromatin modification, and transcription regulation. The study also identified a strong association of TH2B with specific transposable elements (TEs), particularly long terminal repeats (LTRs), which are known to regulate preimplantation development. Additionally, early embryos showed H3K9me3 marks at TH2B-bound loci. TH2B exhibited strong correlations with H2A.Z and H3.3 in the 2-cell and 8-cell stages, a positive association with H3K27Ac and H3K4me3, and a negative correlation with H3K27me3. Allelic reprogramming analysis of TH2B in embryos from C57BL/6J and DBA/2J crosses revealed differential dynamics between maternal and paternal alleles, with a notable paternal bias at the promoter in 2-cell embryos. Thus, TH2B’s enrichment in early embryonic stages and its association with key regulatory regions and histone modifications underscore its importance in ZGA and subsequent developmental processes.

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