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Cell Biochemistry and Function Mar 202450% of cases of infertility are caused by male factor, which acquired or congenital problems may bring on. Male infertility can be caused by oligospermia and...
50% of cases of infertility are caused by male factor, which acquired or congenital problems may bring on. Male infertility can be caused by oligospermia and asthenozoospermia, which are common. Since the same mutations that cause azoospermia in some people also cause oligozoospermia in others, oligozoospermia may be thought of as a less severe form of azoospermia. Studies have demonstrated telomere length, catalase activity, super oxide dismutase (SOD), and DNA fragmentation can be influential factors for male infertility. The amount of apoptosis, oxidative stress factors, telomere length, and DNA fragmentation were some aspects of healthy sperm that we chose to look into in this study and compare to oligospermia individuals. Oligospermia patients (n = 24) and fertile men (n = 27) semen samples were collected, and the apoptosis rate of sperms in both groups was analyzed (Flow cytometry). Also, gene expression of apoptotic and antiapoptotic markers and telomere length were examined (real-time polymerase chain reaction). The sperm DNA fragmentation kit was used to determine DNA fragmentation and to evaluate catalase and SOD activity; the specific kits and methods were utilized. Higher expression levels of caspase3 (p = .0042), caspase8 (p = .0145), caspase9 (p = .0275), and BAX (p = .0202) mRNA were observed in patients who had oligospermia. In contrast, lower mRNA expression of BCL-2 (p = .0009) was detected in this group. In addition, telomere length was decreased in the oligospermia group (p < .0001) compared to the health group. Moreover, the frequency of apoptosis is induced in patients (p = .0026). The catalase activity is low (p = .0008), but the SOD activity is high (p = .0015) in the patient group. As a result of our findings, we may list the sperm cell apoptosis rate, telomere length, the degree of sperm DNA fragmentation, and lastly, the measurement of significant and efficient oxidative stress markers like SOD and catalase in semen plasma among the principal diagnostic characteristics for oligospermia. Future studies will be better able to treat oligospermia by showing whether these indicators are rising or falling.
Topics: Humans; Male; Oligospermia; Reactive Oxygen Species; Catalase; Azoospermia; Semen; Spermatozoa; Infertility, Male; Antioxidants; DNA Fragmentation; Apoptosis; Superoxide Dismutase; Telomere; RNA, Messenger
PubMed: 38379260
DOI: 10.1002/cbf.3935 -
Journal of Medical Genetics May 2024The association between the variants and human infertility remains unclear, as only one homozygous missense variant of was found to be associated with...
BACKGROUND
The association between the variants and human infertility remains unclear, as only one homozygous missense variant of was found to be associated with oligoasthenoteratozoospermia (OAT).
METHODS
Whole-exome sequencing and Sanger sequencing were employed to identify potential pathogenic variants of in infertile men. Histology, immunofluorescence, immunoblotting and ultrastructural analyses were conducted to clarify the structural and functional abnormalities of sperm in mutated patients. -knockout mice were generated using the CRISPR-Cas9 system. Total RNA-seq and single-cell RNA-seq (scRNA-seq) analyses were used to elucidate the underlying molecular mechanisms, followed by validation through quantitative RT-PCR and immunostaining. Intracytoplasmic sperm injection (ICSI) was also used to assess the efficacy of clinical treatment.
RESULTS
Bi-allelic variants were identified in five unrelated Chinese individuals with OAT, including homozygous loss-of-function variants in two consanguineous families. Notably, besides reduced concentrations and impaired motility, a significant occurrence of acrosomal hypoplasia was detected in multiple spermatozoa among five patients. Using the -deficient mice, we further elucidate the pivotal role of TDRD6 in spermiogenesis and acrosome identified. In addition, the mislocalisation of crucial chromatoid body components DDX4 (MVH) and UPF1 was also observed in round spermatids from patients harbouring variants. ScRNA-seq analysis of germ cells from a patient with variants revealed that TDRD6 regulates mRNA metabolism processes involved in spermatid differentiation and cytoplasmic translation.
CONCLUSION
Our findings strongly suggest that TDRD6 plays a conserved role in spermiogenesis and confirms the causal relationship between variants and human OAT. Additionally, this study highlights the unfavourable ICSI outcomes in individuals with bi-allelic variants, providing insights for potential clinical treatment strategies.
Topics: Adult; Animals; Humans; Male; Mice; Acrosome; Alleles; Asthenozoospermia; Exome Sequencing; Infertility, Male; Mice, Knockout; Oligospermia; Pedigree; Sperm Injections, Intracytoplasmic; Spermatogenesis; Spermatozoa
PubMed: 38341271
DOI: 10.1136/jmg-2023-109766 -
International Journal of Molecular... Jan 2024Phosphorus-containing metabolites occupy a prominent position in cell pathways. The phosphorometabolomic approach in human sperm samples will deliver valuable...
Phosphorus-containing metabolites occupy a prominent position in cell pathways. The phosphorometabolomic approach in human sperm samples will deliver valuable information as new male fertility biomarkers could emerge. This study analyzed, by P-NMR, seminal plasma and whole semen from asthenozoospermic and normozoospermic samples (71% vs. 27% and 45% vs. 17%, total and progressive sperm motility, respectively), and also ejaculates from healthy donors. At least 16 phosphorus-containing metabolites involved in central energy metabolism and phospholipid, nucleotide, and nicotinamide metabolic pathways were assigned and different abundances between the samples with distinct sperm quality was detected. Specifically, higher levels of phosphocholine, glucose-1-phosphate, and to a lesser degree, acetyl phosphate were found in the asthenozoospermic seminal plasma. Notably, the phosphorometabolites implicated in lipid metabolism were highlighted in the seminal plasma, while those associated with carbohydrate metabolism were more abundant in the spermatozoa. Higher levels of phosphocholine, glucose-1-phosphate, and acetyl phosphate in the seminal plasma with poor quality suggest their crucial role in supporting sperm motility through energy metabolic pathways. In the seminal plasma, phosphorometabolites related to lipid metabolism were prominent; however, spermatozoa metabolism is more dependent on carbohydrate-related energy pathways. Understanding the presence and function of sperm phosphorylated metabolites will enhance our knowledge of the metabolic profile of healthy human sperm, improving assessment and differential diagnosis.
Topics: Humans; Male; Semen; Phosphorylcholine; Sperm Motility; Spermatozoa; Asthenozoospermia; Phosphorus; Semen Analysis; Organophosphates
PubMed: 38338962
DOI: 10.3390/ijms25031682 -
Human Reproduction (Oxford, England) Apr 2024What is the significance and mechanism of human seminal plasma extracellular vesicles (EVs) in regulating human sperm functions?
STUDY QUESTION
What is the significance and mechanism of human seminal plasma extracellular vesicles (EVs) in regulating human sperm functions?
SUMMARY ANSWER
EV increases the intracellular Ca2+ concentrations [Ca2+]i via extracellular Ca2+ influx by activating CatSper channels, and subsequently modulate human sperm motility, especially hyperactivated motility, which is attributed to both protein and non-protein components in EV.
WHAT IS KNOWN ALREADY
EVs are functional regulators of human sperm function, and EV cargoes from normal and asthenozoospermic seminal plasma are different. Pre-fusion of EV with sperm in the acidic and non-physiological sucrose buffer solution could elevate [Ca2+]i in human sperm. CatSper, a principle Ca2+ channel in human sperm, is responsible for the [Ca2+]i regulation when sperm respond to diverse extracellular stimuli. However, the role of CatSper in EV-evoked calcium signaling and its potential physiological significance remain unclear.
STUDY DESIGN, SIZE, DURATION
EV isolated from the seminal plasma of normal and asthenozoospermic semen were utilized to investigate the mechanism by which EV regulates calcium signal in human sperm, including the involvement of CatSper and the responsible cargoes in EV. In addition, the clinical application potential of EV and EV protein-derived peptides were also evaluated. This is a laboratory study that went on for more than 5 years and involved more than 200 separate experiments.
PARTICIPANTS/MATERIALS, SETTING, METHODS
Semen donors were recruited in accordance with the Institutional Ethics Committee on human subjects of the Affiliated Hospital of Nantong University and Jiangxi Maternal and Child Health Hospital. The Flow NanoAnalyzer, western blotting, and transmission electron microscope were used to systematically characterize seminal plasma EV. Sperm [Ca2+]i responses were examined by fluorimetric measurement. The whole-cell patch-clamp technique was performed to record CatSper currents. Sperm motility parameters were assessed by computer-assisted sperm analysis. Sperm hyperactivation was also evaluated by examining their penetration ability in viscous methylcellulose media. Protein and non-protein components in EV were analyzed by liquid chromatography-mass spectrum. The levels of prostaglandins, reactive oxygen species, malonaldehyde, and DNA integrity were detected by commercial kits.
MAIN RESULTS AND THE ROLE OF CHANCE
EV increased [Ca2+]i via an extracellular Ca2+ influx, which could be suppressed by a CatSper inhibitor. Also, EV potentiated CatSper currents in human sperm. Furthermore, the EV-in [Ca2+]i increase and CatSper currents were absent in a CatSper-deficient sperm, confirming the crucial role of CatSper in EV induced Ca2+ signaling in human sperm. Both proteins and non-protein components of EV contributed to the increase of [Ca2+]i, which were important for the effects of EV on human sperm. Consequently, EV and its cargos promoted sperm hyperactivated motility. In addition, seminal plasma EV protein-derived peptides, such as NAT1-derived peptide (N-P) and THBS-1-derived peptide (T-P), could activate the sperm calcium signal and enhance sperm function. Interestingly, EV derived from asthenozoospermic semen caused a lower increase of [Ca2+]i than that isolated from normal seminal plasma (N-EV), and N-EV significantly improved sperm motility and function in both asthenozoospermic samples and frozen-thawed sperm.
LARGE SCALE DATA
N/A.
LIMITATIONS, REASONS FOR CAUTION
This was an in vitro study and caution must be taken when extrapolating the physiological relevance to in vivo regulation of sperm.
WIDER IMPLICATIONS OF THE FINDINGS
Our findings demonstrate that the CatSper-mediated-Ca2+ signaling is involved in EV-modulated sperm function under near physiological conditions, and EV and their derivates are a novel CatSper and sperm function regulators with potential for clinical application. They may be developed to improve sperm motility resulting from low [Ca2+]i response and/or freezing and thawing.
STUDY FUNDING/COMPETING INTEREST(S)
This research was supported by the National Natural Science Foundation of China (32271167), the Social Development Project of Jiangsu Province (BE2022765), the Nantong Social and People's Livelihood Science and Technology Plan (MS22022087), the Basic Science Research Program of Nantong (JC22022086), and the Jiangsu Innovation and Entrepreneurship Talent Plan (JSSCRC2021543). The authors declare no conflict of interest.
Topics: Humans; Male; Asthenozoospermia; Calcium; Calcium Channels; Calcium Signaling; Peptides; Semen; Sperm Motility; Spermatozoa; Extracellular Vesicles
PubMed: 38335261
DOI: 10.1093/humrep/deae018 -
Journal of Cellular and Molecular... Feb 2024Several studies have highlighted the functional indispensability of methyltransferase-like 3 (METTL3) in the reproductive system. However, a review that comprehensively... (Review)
Review
Several studies have highlighted the functional indispensability of methyltransferase-like 3 (METTL3) in the reproductive system. However, a review that comprehensively interprets these studies and elucidates their relationships is lacking. Therefore, the present work aimed to review studies that have investigated the functions of METTL3 in the reproductive system (including spermatogenesis, follicle development, gametogenesis, reproductive cancer, asthenozoospermia and assisted reproduction failure). This review suggests that METTL3 functions not only essential for normal development, but also detrimental in the occurrence of disorders. In addition, promising applications of METTL3 as a diagnostic or prognostic biomarker and therapeutic target for reproductive disorders have been proposed. Collectively, this review provides comprehensive interpretations, novel insights, potential applications and future perspectives on the role of METTL3 in regulating the reproductive system, which may be a valuable reference for researchers and clinicians.
Topics: Male; Humans; Methyltransferases; RNA; Spermatogenesis; Reproduction; Genitalia
PubMed: 38332508
DOI: 10.1111/jcmm.18128 -
Reports of Biochemistry & Molecular... Jul 2023The influence of cytokine in the reproductive system is becoming increasingly important. The polymorphisms of the transforming growth factor-β1 (TGF-β1) gene are...
BACKGROUND
The influence of cytokine in the reproductive system is becoming increasingly important. The polymorphisms of the transforming growth factor-β1 (TGF-β1) gene are involved in male infertility. This study aimed to demonstrate the association between TGF-β1 and infertility and to investigate its impact on semen quality.
METHODS
In this case-control study, serum TGF-β1 concentration was measured in 144 patients diagnosed with infertility and 40 fertile males by enzyme-linked immunosorbent assay (ELISA). The tetra-amplification refractory mutation system-PCR (T-ARMS-PCR) analysis was performed to detect the genotyping of the TGF-β1 (+869 C/T) (rs1800470) SNPs gene.
RESULTS
Serum concentration of TGF-β1 was less in infertile males compared to fertile ones. The detected and more effective genotypes and alleles of TGF-β1 gene polymorphic on male infertility were, in normozoospermic group, CT genotype, probability (p)= 0.45, relative risk (RR)= 1.56, confidence intervals (CI): 0.58-4.22, and T allele (p= 0.46, RR= 1.32, CI: 0.65-2.69), in oligozoospermic and azoospermic groups, CC genotype (p= 0.32, RR= 1.58, CI: 0.73-3.41), (p= 0.013, RR= 3.50, CI: 1.40-8.73), and allele C (p= 0.44, RR= 1.32, CI: 0.73-2.38), (p= 0.06, RR= 2.14, CI: 1.02-4.50), respectively. The recessive model (TT+CT) showed increased risk among normozoospermic group (p=0.44, RR=1.67, CI:0.60-4.62). The serum concentration of TGF-β1 with CT and TT genotypes was less than that of CC genotype. TGF-β1 C/T genotype correlated with low sperm number, high immotile sperm, and high abnormal sperm morphology.
CONCLUSIONS
Our study revealed that the TGF-β1(rs1800470) gene polymorphisms are associated negatively with semen quality.
PubMed: 38317808
DOI: 10.61186/rbmb.12.2.318 -
Aging Jan 2024Oligoasthenoteratozoospermia (OAT) decreases male fertility, seriously affecting the production of offspring. This study clarified the preventive impact of different...
Oligoasthenoteratozoospermia (OAT) decreases male fertility, seriously affecting the production of offspring. This study clarified the preventive impact of different moxibustion frequencies on OAT and selected the optimal frequency to elucidate the underlying mechanism. An OAT rat model was constructed by gavage of tripterygium glycosides (TGS) suspension. Daily moxibustion (DM) or alternate-day moxibustion (ADM) was administered on the day of TGS suspension administration. Finally, we selected DM for further study based on sperm quality and DNA fragmentation index, testicular and epididymal morphology, and reproductive hormone level results. Subsequently, the oxidative stress (OS) status was evaluated by observing the OS indices levels; malondialdehyde (MDA), 8-hydroxy-deoxyguanosine (8-OHdG), total antioxidant capacity (T-AOC), and total superoxide dismutase (T-SOD) in testicular tissue using colorimetry and enzyme-linked immunosorbent assay. Furthermore, heme oxygenase 1 (HO-1) and nuclear factor erythropoietin-2-related factor 2 (Nrf2) were evaluated using Western blotting. Immunohistochemistry was employed to locate and assess the expression of HO-1 and Nrf2 protein, while quantitative real-time polymerase chain reaction was utilized to detect their mRNA expression. MDA and 8-OHdG levels decreased following DM treatment, while T-SOD and T-AOC increased, suggesting that DM may prevent TGS-induced OAT in rats by decreasing OS in the testis. Furthermore, protein and mRNA expression of Nrf2 and HO-1 in the testis were elevated, indicating that DM may reduce OS by activating the signaling pathway of Nrf2/HO-1. Therefore, DM could prevent OAT in rats via the Nrf2/HO-1 pathway, thereby presenting a promising therapeutic approach against OAT.
Topics: Rats; Male; Animals; Humans; Heme Oxygenase-1; Rats, Sprague-Dawley; NF-E2-Related Factor 2; Tripterygium; Oligospermia; Glycosides; Moxibustion; Asthenozoospermia; Infertility, Male; Seeds; Oxidative Stress; Antioxidants; Signal Transduction; Superoxide Dismutase; RNA, Messenger
PubMed: 38277193
DOI: 10.18632/aging.205475 -
Biomedicines Jan 2024VPS13A, also known as chorein, whose loss of function causes chorea-acanthocytosis (ChAc), is characterized by Huntington's-disease-like neurodegeneration and...
VPS13A, also known as chorein, whose loss of function causes chorea-acanthocytosis (ChAc), is characterized by Huntington's-disease-like neurodegeneration and neuropsychiatric symptoms in addition to acanthocytosis in red blood cells. We previously reported that ChAc-model mice with a loss of chorein function exhibited male infertility, with asthenozoospermia and mitochondrial dysmorphology in the spermatozoa. Here, we report a novel aspect of chorein dysfunction in male fertility, particularly its role in spermatogenesis and mitochondrial integrity. An increase in anti-malondialdehyde antibody immunoreaction within the testes, predominantly observed at the advanced stages of sperm formation in chorein-deficient mice, suggests oxidative stress as a contributing factor to mitochondrial dysfunction and impaired sperm maturation. The chorein immunoreactivity in spermatids of wild-type mice accentuates its significance in sperm development. ChAc-model mice exhibit mitochondrial ultrastructural abnormalities, specifically during the late stages of sperm maturation, suggesting a critical timeframe for chorein's action in spermiogenesis. We observed an increase in TOM20 protein levels, indicative of disrupted mitochondrial import mechanisms. The concurrent decrease in metabolic enzymes such as IDH3A, LDHC, PGK2, and ACAT1 suggests a complex chorein-mediated metabolic network that is essential for sperm vitality. Additionally, heightened separation of cytoplasmic droplets from sperm highlights the potential membrane instability in chorein-deficient spermatozoa. Metabolomic profiling further suggests a compensatory metabolic shift, with elevated glycolytic and TCA-cycle substrates. Our findings suggest that chorein is involved in anti-ferroptosis and the maturation of mitochondrial morphology in the late stages of spermatogenesis, and its deficiency leads to asthenozoospermia characterized by membrane instability, abnormal cytosolic glycolysis, abnormal mitochondrial function, and a disrupted TCA cycle. Further analyses are required to unravel the molecular mechanisms that directly link these findings and to elucidate the role of chorein in spermatogenesis as well as its broader implications.
PubMed: 38275411
DOI: 10.3390/biomedicines12010240 -
Environmental Science and Pollution... Feb 2024Di (2-ethyl-hexyl) phthalate (DEHP) mainly enters the human body through the digestive tract, respiratory tract, and skin. At the same time, it has reproductive and...
Di (2-ethyl-hexyl) phthalate (DEHP) mainly enters the human body through the digestive tract, respiratory tract, and skin. At the same time, it has reproductive and developmental toxicity, neurotoxicity, and so on, which can cause the decrease of sperm motility. Asthenospermia is also known as low sperm motility, and the semen quality of men in some areas of China is declining year by year. Interestingly, previous studies have shown that sleep disorders can also lead to asthenospermia. However, the relationship between sleep, DEHP, and asthenospermia is still unclear. Analysis of the National Health and Nutrition Examination Survey (NHANES) population database showed that DEHP was associated with sleep disorders, and subsequent experiments in mice and Drosophila indicated that DEHP exposure had certain effects on sleep and asthenospermia. Furthermore, we analyzed the Comparative Toxicogenomics Database (CTD) to find out the common signaling pathway among the three: hypoxia-inducible factor 1(HIF-1). Then Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) was used to screen out the proteins that DEHP affected the HIF-1 pathway: glyceraldehyde-3-phosphate dehydrogenase (GAPDH), serine/threonine-protein kinase (AKT1), epidermal growth factor receptor (EGFR), and finally Western blot analysis was used to detect the expression levels of the three proteins. Compared with the control group, DEHP decreased the protein expression levels of GAPDH and AKT1 in the HIF-1 pathway, and caused sleep disorders and decreased sperm motility. This study provides preliminary evidence for exploring the mechanism among DEHP, sleep disorders, and asthenospermia.
Topics: Humans; Male; Animals; Mice; Diethylhexyl Phthalate; Semen Analysis; Nutrition Surveys; Sperm Motility; Sleep; Sleep Wake Disorders; Phthalic Acids
PubMed: 38265591
DOI: 10.1007/s11356-024-32030-9 -
BMC Microbiology Jan 2024Identification of intestinal flora composition is significant for exploring the cause and pathogenic mechanisms of the gut-testis axis and clarifying the relationship...
BACKGROUND
Identification of intestinal flora composition is significant for exploring the cause and pathogenic mechanisms of the gut-testis axis and clarifying the relationship between microbiota and infertility. Our study aimed to examine the alternation in gut microbiota composition and identify potential microbes associated with development of Asthenozoospermia (AS).
METHOD
A total of 580 males were recruited in the outpatient department of Tianjin Medical University General Hospital between September 2021 and March 2023. Sperm parameters were analyzed according to the WHO laboratory manual. The 16 S rRNA gene high-throughput sequencing was performed to detect the gut microbiota composition in fecal samples. LEfSe analysis was used to screen key microbiota. PICRUSt2 software was utilized to predict relevant pathways.
RESULTS
After rigorous screening, 60 isolated AS patients (AS group) and 48 healthy men (NC group) were enrolled. No significant differences were observed in demographic characteristics (p > 0.05), semen volume (p = 0.718), sperm concentration (p = 0.109), or total sperm count (p = 0.200). Sperm total motility and progressive motility were significantly decreased in the AS group (p < 0.001). AS patients had significantly lower alpha diversity indices (Chao1, observed OTUs, and PD Whole-tree; p < 0.05). The beta-diversity of gut microbiota in AS patients significantly differed from NC men (PCoA analysis, p = 0.001). Firmicutes, Bacteroidota, Proteobacteria, and Actinobacteria were the primary phyla, with the dominant genera including Bacteroides, Prevotella, and Blautia. Eleven key genera such as Escherichia_Shigella and Prevotellaceae_UCG_001 were identified by LEfSe analysis. Most of these genera were negatively correlated with sperm mobility. Eighty-eight KEGG pathways, including steroid biosynthesis and meiosis, were significantly enriched between the two groups.
CONCLUSIONS
It appears that gut microbiota composition in AS patients significantly differed from that in healthy men, and the development of AS might be associated with intestinal flora dysbiosis.
Topics: Humans; Male; Gastrointestinal Microbiome; Pilot Projects; Asthenozoospermia; Semen; Bacteroidetes; Dysbiosis; China; RNA, Ribosomal, 16S
PubMed: 38225541
DOI: 10.1186/s12866-023-03173-5