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Asian Journal of Andrology Apr 2023Phosphodiesterase (PDE) inhibitors can improve sperm motility in patients with asthenozoospermia. However, the most commonly reported nonselective PDE inhibitor...
Phosphodiesterase (PDE) inhibitors can improve sperm motility in patients with asthenozoospermia. However, the most commonly reported nonselective PDE inhibitor pentoxifylline and PDE5 inhibitor sildenafil have the disadvantages of requiring a high concentration and destroying sperm integrity. We examined the PDE10A inhibitor PF-2545920 to compare its ability to promote sperm motility with that of pentoxifylline and sildenafil. After seminal plasma was discarded, several semen samples were subjected to four treatments (control, PF-2545920, pentoxifylline, and sildenafil) to evaluate their ability to affect motility, viability, and spontaneous acrosome reactions. Intracellular calcium and adenosine triphosphate (ATP), mitochondrial membrane potential, and penetration through viscous medium were assessed by flow cytometry, luciferase, and hyaluronic acid after treatment with PF-2545920. Statistical analyses were performed using the analysis of variance statistical test. PF-2545920 elevated the percentage of motile spermatozoa compared to the control, pentoxifylline, and sildenafil groups at 10 µmol l -1 ( P < 0.01). It is less toxic to GC-2spd mouse spermatocytes cells and spermatozoa and causes fewer spontaneous acrosomal reactions ( P < 0.05). PF-2545920 also increased mitochondrial membrane potential ( P < 0.001) and altered intracellular calcium ( P < 0.05) in a dose-dependent manner, including increasing sperm hyaluronic acid penetrating ability ( P < 0.05). Therefore, PF-2545920 might be an excellent choice for stimulating the sperm motility.
PubMed: 37026191
DOI: 10.4103/aja2022117 -
Sensors (Basel, Switzerland) Feb 2023(kinetochore scaffold 1) has attracted much attention as one of the assembly elements of the outer kinetochore, and the functions of its different domains have been...
The Loss-Function of Causes Oligospermia and Asthenospermia in Mice by Affecting the Assembly and Separation of the Spindle through Flow Cytometry and Immunofluorescence.
(kinetochore scaffold 1) has attracted much attention as one of the assembly elements of the outer kinetochore, and the functions of its different domains have been gradually revealed, most of which are associated with cancers, but few links have been made between and male fertility. Here, we first linked to male reproductive health and the loss-function of resulted in oligospermia and asthenospermia in mice (an 86.5% decrease in total sperm number and an 82.4% increase in static sperm number, respectively) through CASA (computer-aided sperm analysis). Moreover, we introduced an ingenious method to pinpoint the abnormal stage in the spermatogenic cycle using flow cytometry combined with immunofluorescence. Results showed that 49.5% haploid sperm was reduced and 53.2% diploid sperm was increased after the function of was lost. Spermatocytes arrest was identified at the meiotic prophase I of spermatogenesis, which was induced by the abnormal assembly and separation of the spindle. In conclusion, we established an association between and male fertility, providing a guide for future genetic counseling regarding oligospermia and asthenospermia, and a powerful method for further exploring spermatogenic dysfunction by utilizing flow cytometry and immunofluorescence.
Topics: Animals; Male; Mice; Asthenozoospermia; Flow Cytometry; Fluorescent Antibody Technique; Meiosis; Oligospermia; Semen; Microtubule-Associated Proteins
PubMed: 36904774
DOI: 10.3390/s23052571 -
Asian Journal of Andrology Mar 2023We examined a cohort of 93 cystic fibrosis (CF) male patients who were pancreatic-sufficient (PS-CF; n=40) or pancreatic-insufficient (PI-CF; n = 53). Complex semen...
We examined a cohort of 93 cystic fibrosis (CF) male patients who were pancreatic-sufficient (PS-CF; n=40) or pancreatic-insufficient (PI-CF; n = 53). Complex semen examination was performed, including standard semen analysis, quantitative karyological analysis (QKA) of immature germ cells (IGCs), transmission electronic microscopy (TEM), biochemical analysis, and sperm DNA fragmentation by terminal deoxynucleotidyl transferase-mediated dUTP nickend labeling (TUNEL) assay. Azoospermia was diagnosed in 83 (89.2%) patients. The other 10 (10.8%) patients were found to be nonazoospermic and showed various spermatological diagnoses (asthenozoospermia, n = 2; asthenoteratozoospermia, n = 3; oligoasthenozoospermia, n = 1; oligoasthenoteratozoospermia, n = 3; and normozoospermia, n = 1) with no specific morphological abnormalities. Oligospermia was detected in 89.2% azoospermic and 30.0% nonazoospermic patients. Low seminal pH (<7.0) was found in 74 (89.2%) of 83 azoospermic patients. Moderate leukocytospermia (2.0 × 10 6 -2.2 × 10 6 ml -1 ) was revealed in 2.4% azoospermic and 40.0% nonazoospermic semen samples. The signs of partial meiotic arrest at prophase I were found in 4 of 6 nonazoospermic patients examined by QKA of IGCs. The content of fructose and citrate was low in oligospermic and normal in nonoligospermic semen samples. An increased percentage (>30%) of spermatozoa with noncondensed ("immature") chromatin was revealed in 2 of 6 nonazoospermic semen samples analyzed by TEM.
PubMed: 36891936
DOI: 10.4103/aja2022115 -
Frontiers in Genetics 2023Primary Ciliary Dyskinesia (PCD) is a rare genetic disorder affecting the function of motile cilia in several organ systems. In PCD, male infertility is caused by...
Primary Ciliary Dyskinesia (PCD) is a rare genetic disorder affecting the function of motile cilia in several organ systems. In PCD, male infertility is caused by defective sperm flagella composition or deficient motile cilia function in the efferent ducts of the male reproductive system. Different PCD-associated genes encoding axonemal components involved in the regulation of ciliary and flagellar beating are also reported to cause infertility due to multiple morphological abnormalities of the sperm flagella (MMAF). Here, we performed genetic testing by next generation sequencing techniques, PCD diagnostics including immunofluorescence-, transmission electron-, and high-speed video microscopy on sperm flagella and andrological work up including semen analyses. We identified ten infertile male individuals with pathogenic variants in (one) and (two) encoding ruler proteins, (two) and (one) encoding radial spoke head proteins, and (two) and (two) encoding CP-associated proteins, respectively. We demonstrate for the first time that pathogenic variants in and cause male infertility due to sperm cell dysmotility and abnormal flagellar RSPH1 and RSPH9 composition. We also provide novel evidence for MMAF in - and -mutant individuals. We show absence or severe reduction of CCDC39 and SPEF2 in sperm flagella of - and -mutant individuals and - and -mutant individuals, respectively. Thereby, we reveal interactions between CCDC39 and CCDC40 as well as HYDIN and SPEF2 in sperm flagella. Our findings demonstrate that immunofluorescence microscopy in sperm cells is a valuable tool to identify flagellar defects related to the axonemal ruler, radial spoke head and the central pair apparatus, thus aiding the diagnosis of male infertility. This is of particular importance to classify the pathogenicity of genetic defects, especially in cases of missense variants of unknown significance, or to interpret variants that are confounded by the presence of the almost identical pseudogene .
PubMed: 36873931
DOI: 10.3389/fgene.2023.1117821 -
Developmental Biology May 2023Male infertility affects approximately 7% of childbearing couples and is a major health issue. Although nearly 50% idiopathic infertile men are assumed to have a genetic...
Male infertility affects approximately 7% of childbearing couples and is a major health issue. Although nearly 50% idiopathic infertile men are assumed to have a genetic basis, the underlying causes remain largely unknown in most infertility cases. Here, we report two rare homozygous variants in two previously uncharacterized genes, C9orf131 and C10orf120, identified in two unrelated men with asthenozoospermia. Both genes were predominantly expressed in the testes. Furthermore, C9orf131 and C10orf120 knockout mice were successfully generated using the CRISPR-Cas9 technology. However, both C9orf131 and C10orf120 adult male mice were fertile, with testis-to-body weight ratios comparable to those of wild-type mice. No overt differences were found between wild-type, C9orf131, and C10orf120 mice regarding testicular/epididymal tissue morphology, sperm count, sperm motility, or sperm morphology. Moreover, TUNEL assays indicated that the number of apoptotic germ cells in testes was not significantly different between the three groups. In summary, these findings suggest that C9orf131 and C10orf120 are redundant genes in male infertility.
Topics: Fertility; Humans; Mice; Asthenozoospermia; Mice, Knockout; Testis; Male; Sperm Motility; Sperm Count; Spermatozoa; In Situ Nick-End Labeling; Animals
PubMed: 36871790
DOI: 10.1016/j.ydbio.2023.02.009 -
Genes Feb 2023Infertility is a major health problem worldwide without an effective therapy or cure. It is estimated to affect 8-12% of couples in the reproductive age group, equally... (Review)
Review
Infertility is a major health problem worldwide without an effective therapy or cure. It is estimated to affect 8-12% of couples in the reproductive age group, equally affecting both genders. There is no single cause of infertility, and its knowledge is still far from complete, with about 30% of infertile couples having no cause identified (named idiopathic infertility). Among male causes of infertility, asthenozoospermia (i.e., reduced sperm motility) is one of the most observed, being estimated that more than 20% of infertile men have this condition. In recent years, many researchers have focused on possible factors leading to asthenozoospermia, revealing the existence of many cellular and molecular players. So far, more than 4000 genes are thought to be involved in sperm production and as regulators of different aspects of sperm development, maturation, and function, and all can potentially cause male infertility if mutated. In this review, we aim to give a brief overview of the typical sperm flagellum morphology and compile some of the most relevant information regarding the genetic factors involved in male infertility, with a focus on sperm immotility and on genes related to sperm flagellum development, structure, or function.
Topics: Male; Humans; Female; Sperm Tail; Asthenozoospermia; Semen; Sperm Motility; Infertility, Male
PubMed: 36833310
DOI: 10.3390/genes14020383 -
International Journal of Reproductive... Dec 2022Sperm freezing is an important procedure in assisted reproductive technology. Freezing results in physical and chemical changes in the sperm. () is a tree that has...
BACKGROUND
Sperm freezing is an important procedure in assisted reproductive technology. Freezing results in physical and chemical changes in the sperm. () is a tree that has antioxidant properties.
OBJECTIVE
This study aimed to investigate the effect of different concentrations of in a freezing medium on semen parameters, and some biochemical parameters in asthenozoospermic specimens.
MATERIALS AND METHODS
Forty asthenozoospermic specimens (semen specimens with motility 32%) were obtained from men aged between 20-40 yr according to the World Health Organization criteria. Each sample was divided into 6 groups: I) fresh, II) control, III) 5, IV) 10, V) 20, and VI) 30 µg/ml extract were added to a freezing medium respectively. Then sperm parameters, malondialdehyde, total antioxidant capacity, reactive oxygen species, and sperm DNA assay were evaluated using related protocols after thawing.
RESULTS
Data analysis shows that sperm parameter, and total antioxidant capacity level increased at a concentration of 20 µg/ml of extract compared to the other concentrations of extract after cryopreservation and thawing (p 0.001). Also, the sperm DNA fragmentation assay, reactive oxygen species, and malondialdehyde levels were significantly reduced by adding 20 µg/ml of extract to the sperm freezing medium compared to the other treated groups after cryopreservation (p 0.001).
CONCLUSION
extract significantly improved sperm parameters after cryopreservation and thawing in asthenozoospermic specimens, and the greatest impact was observed at the 20 µg/ml extract concentration (p 0.001).
PubMed: 36819209
DOI: 10.18502/ijrm.v20i12.12564 -
International Journal of Reproductive... Dec 2022Infertility is considered as a common problem appears in about 10-12% of couples in their reproductive ages Ring finger protein 38 gene is a ubiquitin-protein ligase...
BACKGROUND
Infertility is considered as a common problem appears in about 10-12% of couples in their reproductive ages Ring finger protein 38 gene is a ubiquitin-protein ligase that can regulate Protein 53 and affect cellular motility.
OBJECTIVE
Considering the role of on cellular motility and on the regulation of , the present study aimed to assess the difference between and genes expression in normozoospermic and asthenospermic samples as a diagnostic biomarker in males.
MATERIALS AND METHODS
The present study was conducted among 21 asthenospermicsand 63 healthy individuals. First, the real-time polymerase chain reaction technique was applied to measure the expression level of the and genes extracted from sperm samples, and the glyceraldehyde-3phosphate dehydrogenase gene was selected as the reference gene.
RESULTS
An increase and a decrease occurred in the level of and genes expressions in asthenospermic and normozoospermic samples, respectively. In addition, a significant difference was observed between increasing gene expression (p 0.001), reducing one, and decreasing sperm motility (p 0.001) in asthenospermic cells compared to that of normozoospermic ones.
CONCLUSION
Based on the results, an increase in the expression of the gene and a decrease in the expression of the gene had a significant relationship with asthenospermia in men. Therefore, it is expected that an effective step should be adopted to diagnose the asthenospermia expression pattern by using these results.
PubMed: 36819206
DOI: 10.18502/ijrm.v20i12.12563 -
Scientific Reports Feb 2023Asthenozoospermia (AZS) is a severe form of male infertility with no clear pathogenesis, despite numerous research efforts, there is no consensus on this. This study was...
Asthenozoospermia (AZS) is a severe form of male infertility with no clear pathogenesis, despite numerous research efforts, there is no consensus on this. This study was to investigate the expression of gene-associated with retinoid-interferon-induced mortality 19 (GRIM-19) in the sperm of patients with asthenozoospermia and the regulation of GC-2 spd cell proliferation, apoptosis and migration. We analyzed the sperm samples from 82 asthenozoospermia and normal patients were collected in the First People's Hospital of Shangqiu and the First Affiliated Hospital of Zhengzhou University. Immunofluorescence, western blots and RT-qPCR analyses were used to verify the expressions of GRIM-19. MTT assays were used to assess cell proliferations, flow cytometry was performed to assess cell apoptosis, wound‑healing was performed to measure cell migration. Immunofluorescence showed that GRIM-19 is predominantly expressed in the sperm mid-piece, the mRNA expressions of GRIM-19 in sperms of the asthenozoospermia group were significantly low, relative to the normal group (OR 0.266; 95% CI = 0.081-0.868; P = 0.028). The protein expressions of GRIM-19 in sperms of the asthenozoospermia group were significantly lower than that of the normal group as well (GRIM-19/GAPDH: 0.827 ± 0.063 vs 0.458 ± 0.033; P < 0.001). GRIM-19 overexpression promotes GC-2 spd cell proliferation and migration and reduces apoptosis, while GRIM-19-silenced reduces GC-2 spd cell proliferation and migration and increased apoptosis. GRIM-19 is closely related to the occurrence of asthenozoospermia and promotes GC-2 spd cell proliferation and migration and reduces apoptosis.
Topics: Humans; Male; Asthenozoospermia; Semen; Apoptosis; Apoptosis Regulatory Proteins; Cell Proliferation
PubMed: 36813832
DOI: 10.1038/s41598-023-29775-7 -
American Journal of Human Genetics Mar 2023Primate-specific genes (PSGs) tend to be expressed in the brain and testis. This phenomenon is consistent with brain evolution in primates but is seemingly contradictory...
Primate-specific genes (PSGs) tend to be expressed in the brain and testis. This phenomenon is consistent with brain evolution in primates but is seemingly contradictory to the similarity of spermatogenesis among mammals. Here, using whole-exome sequencing, we identified deleterious variants of X-linked SSX1 in six unrelated men with asthenoteratozoospermia. SSX1 is a PSG expressed predominantly in the testis, and the SSX family evolutionarily expanded independently in rodents and primates. As the mouse model could not be used for studying SSX1, we used a non-human primate model and tree shrews, which are phylogenetically similar to primates, to knock down (KD) Ssx1 expression in the testes. Consistent with the phenotype observed in humans, both Ssx1-KD models exhibited a reduced sperm motility and abnormal sperm morphology. Further, RNA sequencing indicated that Ssx1 deficiency influenced multiple biological processes during spermatogenesis. Collectively, our experimental observations in humans and cynomolgus monkey and tree shrew models highlight the crucial role of SSX1 in spermatogenesis. Notably, three of the five couples who underwent intra-cytoplasmic sperm injection treatment achieved a successful pregnancy. This study provides important guidance for genetic counseling and clinical diagnosis and, significantly, describes the approaches for elucidating the functions of testis-enriched PSGs in spermatogenesis.
Topics: Animals; Male; Asthenozoospermia; Macaca fascicularis; Primates; Semen; Sperm Motility; Tupaia; Tupaiidae
PubMed: 36796361
DOI: 10.1016/j.ajhg.2023.01.016