Project description:Currently, the molecular mechanisms for the cause of male infertility are still poorly understood. Our previous study has demonstrated that PIWI-interacting RNAs (piRNAs) are downregulated in seminal plasma of infertile patients and can serve as molecular biomarkers for male infertility. However, the source and mechanism for the dysregulation of piRNAs remain obscure. In this study, we found that exosomes are present in high concentrations in human seminal plasma and confirmed that piRNAs are predominantly present in the exosomal fraction of seminal plasma. Moreover, we showed that piRNAs were significantly decreased in exosomes of asthenozoospermia patients compared with the fertile controls. By systematically screening piRNA profiles in sperms of fertile controls and asthenozoospermia patients, we found that piRNAs were parallelly reduced during infertility. At last, we investigated the expression of proteins that are essential for piRNAs biogenesis in sperms and therefore identified a tight correlation between the levels of spermatozoa piRNA and MitoPLD protein, suggesting that the loss-of-function of MitoPLD could cause a severe defect of piRNA accumulation in sperms. In summary, this study identified a parallel reduction of piRNAs and MitoPLD protein in sperms of infertile patients, which may provide pathophysiological clues about the development of infertility.
Project description:Although piwi-interacting RNAs (piRNAs) play pivotal roles in spermatogenesis, little is known about piRNAs in the seminal plasma of infertile males. In this study, we systematically investigated the profiles of seminal plasma piRNAs in infertile males to identify piRNAs that are altered during infertility and evaluate their diagnostic value. Seminal plasma samples were obtained from 211 infertile patients (asthenozoospermia and azoospermia) and 91 fertile controls. High-throughput sequencing technology was employed to screen piRNA profiles in seminal plasma samples pooled from healthy controls and infertile patients. The results identified 61 markedly altered piRNAs in the infertile patient groups compared with the control group. Next, a quantitative RT-PCR assay was conducted in the training and validation sets to measure and confirm the concentrations of altered piRNAs. The results identified a panel of 5 piRNAs that were significantly decreased in the seminal plasma of infertile patients compared with healthy controls. The areas under the ROC curves for these piRNAs ranged from 0.796 to 0.996, suggesting that the diagnostic potential of these 5 piRNAs to distinguish asthenozoospermic and azoospermic individuals from healthy controls was high. In summary, this study identifies a panel of piRNAs that can accurately distinguish fertile from infertile males. This finding may provide pathophysiological clues that are involved in the development of infertility. Fresh samples were collected and stored at -80â??.Total RNA of seminal plasma were extracted and solexa sequencing was performed.
Project description:Although piwi-interacting RNAs (piRNAs) play pivotal roles in spermatogenesis, little is known about piRNAs in the seminal plasma of infertile males. In this study, we systematically investigated the profiles of seminal plasma piRNAs in infertile males to identify piRNAs that are altered during infertility and evaluate their diagnostic value. Seminal plasma samples were obtained from 211 infertile patients (asthenozoospermia and azoospermia) and 91 fertile controls. High-throughput sequencing technology was employed to screen piRNA profiles in seminal plasma samples pooled from healthy controls and infertile patients. The results identified 61 markedly altered piRNAs in the infertile patient groups compared with the control group. Next, a quantitative RT-PCR assay was conducted in the training and validation sets to measure and confirm the concentrations of altered piRNAs. The results identified a panel of 5 piRNAs that were significantly decreased in the seminal plasma of infertile patients compared with healthy controls. The areas under the ROC curves for these piRNAs ranged from 0.796 to 0.996, suggesting that the diagnostic potential of these 5 piRNAs to distinguish asthenozoospermic and azoospermic individuals from healthy controls was high. In summary, this study identifies a panel of piRNAs that can accurately distinguish fertile from infertile males. This finding may provide pathophysiological clues that are involved in the development of infertility.
Project description:The global infertility rate has been increasing annually, and male infertility becoming severe. The proportion of idiopathic infertility is high, with most patients with ambiguous clinical symptoms. These symptoms manifested as declining semen quality, and the effects of autoimmune factors on this measure of fertility cannot be ignored. Aside from anti-sperm antibody for autoimmune infertility, detection indexes for male infertility are lacking. We successfully collected seminal fluid via electric stimulation from an experimental model of autoimmune orchitis in rats. We used the most advanced isobaric tags for relative and absolute quantitative proteomics analysis to analyze the seminal plasma of autoimmune infertility.
Project description:The study of male infertility after spinal cord injury (SCI) has enhanced the understanding of seminal plasma (SP) as an important regulator of spermatozoa function. However, the most important factors leading to the diminished sperm motility and viability observed in semen of men with SCI remained unknown. Thus, to explore SP related molecular mechanisms underlying infertility after SCI we used mass spectrometry-based quantitative proteomics to compare SP retrieved from SCI patients to normal controls. As a result, we present an in-depth characterization of the human SP proteome, identifying ~2,800 unique proteins, and describe, in detail, the differential proteome observed in SCI. Our analysis demonstrates that a hyper-activation of the immune system may influence some seminal processes, which probably are not triggered by microbial infection. Moreover, we show evidence of an important prostate gland functional failure, i.e. diminished abundance of metabolic enzymes related to ATP turnover, secreted via prostasomes and identify the main outcome related to this fact and that it is intrinsically linked to the low sperm motility in SCI. Together, our data suggest the molecular pathways hindering fertility in SCI and shed new light on other causes of male infertility.
Project description:The human seminal plasma is a potential source of biomarkers for male reproductive disorders. A tissue-profiling analysis of the main organs participating in the secretion of this body fluid was conducted to identify tissue-specific genes along the male reproductive tract.
Project description:We found that differentially expressed (DE) miRNAs were mainly enriched in pathways involved in cell apoptosis, including mTOR and p53 signaling pathways. Furthermore, gain- and loss-of-function analyses and dual luciferase assays demonstrated that endogenous miR-26a-5p and let-7g-5p have potential anti-apoptotic and pro-survival functions in sperm cells through targeting PTEN and PMAIP1 genes. Further comparisons revealed high similarities in miRNA profiles between seminal plasma exosomes and sperm cells, both in HC and CT groups. Our study revealed that miRNAs in sperm cells and seminal plasma exosomes have important functions in male reproduction, suggesting they could be used as potential treatment targets or novel diagnostic markers for male infertility.
Project description:The human seminal plasma is a potential source of biomarkers for male reproductive disorders. A tissue-profiling analysis of the main organs participating in the secretion of this body fluid was conducted to identify tissue-specific genes along the male reproductive tract. Total RNA from non pathological Human seminal vesicles were extracted and hybridized on Affymetrix microarrays. Expression signals in seminal vesicles (present dataset), prostates (GEO; GSE7307), epidydimises (GEO; GSE7808) and testicular samples (Arrayexpress; E-TABM-130) were compared to identify genes that are detected in one of these organs only.