RESUMEN

Infertility represents a significant health concern, with sperm quantity and quality being crucial determinants of male fertility. Oligoasthenoteratozoospermia (OAT) is characterized by reduced sperm motility, lower sperm concentration, and morphological abnormalities in sperm heads and flagella. Although variants in several genes have been implicated in OAT, its genetic etiologies and pathogenetic mechanisms remain inadequately understood. In this study, we identified a homozygous nonsense mutation (c.916C>T, p.Arg306*) in the coiled-coil domain containing 146 ( CCDC146) gene in an infertile male patient with OAT. This mutation resulted in the production of a truncated CCDC146 protein (amino acids 1-305), retaining only two out of five coiled-coil domains. To validate the pathogenicity of the CCDC146 mutation, we generated a mouse model ( Ccdc146 mut/mut ) with a similar mutation to that of the patient. Consistently, the Ccdc146 mut/mut mice exhibited infertility, characterized by significantly reduced sperm counts, diminished motility, and multiple defects in sperm heads and flagella. Furthermore, the levels of axonemal proteins, including DNAH17, DNAH1, and SPAG6, were significantly reduced in the sperm of Ccdc146 mut/mut mice. Additionally, both human and mouse CCDC146 interacted with intraflagellar transport protein 20 (IFT20), but this interaction was lost in the mutated versions, leading to the degradation of IFT20. This study identified a novel deleterious homozygous nonsense mutation in CCDC146 that causes male infertility, potentially by disrupting axonemal protein transportation. These findings offer valuable insights for genetic counseling and understanding the mechanisms underlying CCDC146 mutant-associated infertility in human males.

Asunto(s)

Astenozoospermia , Proteínas Asociadas a Microtúbulos , Animales , Humanos , Masculino , Ratones , Astenozoospermia/genética , Codón sin Sentido , Homocigoto , Infertilidad Masculina/genética , Mutación , Oligospermia/genética , Motilidad Espermática/genética , Espermatozoides , Proteínas Asociadas a Microtúbulos/genética

RESUMEN

Male infertility due to asthenozoospermia is quite frequent, but its etiology is poorly understood. We recruited two infertile brothers, born to first-cousin parents from Pakistan, displaying idiopathic asthenozoospermia with mild stuttering disorder but no ciliary-related symptoms. Whole-exome sequencing identified a splicing variant (c.916+1G>A) in ARMC3, recessively co-segregating with asthenozoospermia in the family. The ARMC3 protein is evolutionarily highly conserved and is mostly expressed in the brain and testicular tissue of human. The ARMC3 splicing mutation leads to the exclusion of exon 8, resulting in a predicted truncated protein (p.Glu245_Asp305delfs*16). Quantitative real-time PCR revealed a significant decrease at mRNA level for ARMC3 and Western blot analysis did not detect ARMC3 protein in the patient's sperm. Individuals homozygous for the ARMC3 splicing variant displayed reduced sperm motility with frequent morphological abnormalities of sperm flagella. Transmission electron microscopy of the affected individual IV: 2 revealed vacuolation in sperm mitochondria at the midpiece and disrupted flagellar ultrastructure in the principal and end piece. Altogether, our results indicate that this novel homozygous ARMC3 splicing mutation destabilizes sperm flagella and leads to asthenozoospermia in our patients, providing a novel marker for genetic counseling and diagnosis of male infertility.

Asunto(s)

Astenozoospermia , Consanguinidad , Homocigoto , Linaje , Empalme del ARN , Cola del Espermatozoide , Adulto , Humanos , Masculino , Astenozoospermia/genética , Astenozoospermia/patología , Secuenciación del Exoma , Infertilidad Masculina/genética , Infertilidad Masculina/patología , Mutación , Empalme del ARN/genética , Motilidad Espermática/genética , Cola del Espermatozoide/patología , Cola del Espermatozoide/ultraestructura , Cola del Espermatozoide/metabolismo , Espermatozoides/ultraestructura , Espermatozoides/patología

RESUMEN

BACKGROUND: Bi-allelic variants in DNAH11 have been identified as causative factors in Primary Ciliary Dyskinesia, leading to abnormal respiratory cilia. Nonetheless, the specific impact of these variants on human sperm flagellar and their involvement in male infertility remain largely unknown. METHODS: A collaborative effort involving two Chinese reproductive centers conducted a study with 975 unrelated infertile men. Whole-exome sequencing was employed for variant screening, and Sanger sequencing confirmed the identified variants. Morphological and ultrastructural analyses of sperm were conducted using Scanning Electron Microscopy and Transmission Electron Microscopy. Western Blot Analysis and Immunofluorescence Analysis were utilized to assess protein levels and localization. ICSI was performed to evaluate its efficacy in achieving favorable pregnancy outcomes for individuals with DNAH11 variants. RESULTS: In this study, we identified seven novel variants in the DNAH11 gene in four asthenoteratozoospermia subjects. These variants led the absence of DNAH11 proteins and ultrastructure defects in sperm flagella, particularly affecting the outer dynein arms (ODAs) and adjacent structures. The levels of ODA protein DNAI2 and axoneme related proteins were down regulated, instead of inner dynein arms (IDA) proteins DNAH1 and DNAH6. Two out of four individuals with DNAH11 variants achieved clinical pregnancies through ICSI. The findings confirm the association between male infertility and bi-allelic deleterious variants in DNAH11, resulting in the aberrant assembly of sperm flagella and contributing to asthenoteratozoospermia. Importantly, ICSI emerges as an effective intervention for overcoming reproductive challenges caused by DNAH11 gene variants.

Asunto(s)

Astenozoospermia , Dineínas Axonemales , Secuenciación del Exoma , Infertilidad Masculina , Humanos , Masculino , Astenozoospermia/genética , Astenozoospermia/patología , Dineínas Axonemales/genética , Femenino , Infertilidad Masculina/genética , Infertilidad Masculina/patología , Adulto , Cola del Espermatozoide/patología , Cola del Espermatozoide/ultraestructura , Cola del Espermatozoide/metabolismo , Inyecciones de Esperma Intracitoplasmáticas , Embarazo , Espermatozoides/ultraestructura , Espermatozoides/patología , Dineínas/genética

RESUMEN

Diabetes mellitus (DM) is a chronic metabolic disease that predisposes to chronic damage and dysfunction of various organs, including leading to erectile dysfunction (ED) and asthenospermia. Literature suggests that ginseng plays an important role in the treatment and management of DM. Ginseng may have a therapeutic effect on the complications of DM-induced ED and asthenospermia. The study aimed to explore the mechanisms of ginseng in the treatment of DM-induced ED and asthenospermia following the Traditional Chinese Medicine (TCM) theory of "treating different diseases with the same treatment." This study used network pharmacology and molecular docking to examine the potential targets and pharmacological mechanism of Ginseng for the treatment of DM-induced ED and asthenospermia. The chemical ingredients and targets of ginseng were acquired using the Traditional Chinese Medicine Systems Pharmacology database and analysis platform. The targets of DM, ED, and asthenospermia were extracted with the GeneCards and Online Mendelian Inheritance in Man databases. A protein-protein interaction network analysis was constructed. The Metascape platform was applied for analyzing the gene ontology and Kyoto Encyclopedia of Genes and Genomes pathways. AutoDock Vina was used to perform molecular docking. Network pharmacology revealed that the main active components of the target of action were kaempferol, beta-sitosterol, ginsenoside rh2, stigmasterol, and fumarine. Core targets of the protein-protein interaction network included TNF, IL-1ß, AKT1, PTGS2, BCL2, and JUN. Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that they were mainly involved in AGE-RAGE signaling pathway in diabetic complications, TNF signaling pathway, Lipid and atherosclerosis. The interactions of core active components and targets were analyzed by molecular docking. Ginseng may play a comprehensive therapeutic role in the treatment of DM-induced ED and asthenospermia through "multicomponent, multi-target, and multi-pathway" biological mechanisms such as inflammation and oxidative stress.

Asunto(s)

Astenozoospermia , Disfunción Eréctil , Simulación del Acoplamiento Molecular , Farmacología en Red , Panax , Masculino , Humanos , Panax/química , Disfunción Eréctil/tratamiento farmacológico , Astenozoospermia/tratamiento farmacológico , Medicina Tradicional China/métodos , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéutico , Mapas de Interacción de Proteínas , Complicaciones de la Diabetes/tratamiento farmacológico , Diabetes Mellitus/tratamiento farmacológico , Sitoesteroles/farmacología

RESUMEN

BACKGROUND: Male infertility due to spermatogenesis defects affects millions of men worldwide. However, the genetic etiology of the vast majority remains unclear. The present study was undertaken to assess the association of DNAH6 and ATPase6 genes in asthenozoospermia patients in the northern region of India. METHODS: A total of 60 semen samples were collected for the study, of which 30 were from the case group and 30 were from the control group. The semen samples for the case group (asthenozoospermia) and control groups were collected from IVF and Reproductive Biology Centre, Maulana Azad Medical College, New Delhi. Sperm count and motility were classified as per World Health Organization (WHO 2021) protocol. A total genomic DNA was extracted as per the stranded TRIZOL method with little modification. RESULTS: In-vitro molecular characterizations of DNAH6 and ATPase6 genes in both groups were checked by Polymerase Chain Reaction (PCR). The 675 bp and 375 bp amplicons were amplified using PCR for ATPase6 and DNAH6 genes. Our study results showed a significant (P ≤ 0.05) null deletion of DNAH6 and ATPase6 genes in asthenozoospermia patients as compared to the control. We found the significant null deletion of DNAH6 in case 45.0%, and the control group was 11.7%. However, in the case of APTase6, it was 26.7% and 10.0%, respectively. CONCLUSIONS: Our study concluded that the presence of DHAH6 and ATPase6 genes had a significant impact on male infertility.

Asunto(s)

Astenozoospermia , Humanos , Masculino , Astenozoospermia/genética , India , Adulto , ATPasas de Translocación de Protón Mitocondriales/genética , ADN Mitocondrial/genética

RESUMEN

Among the many factors with a proven relation to semen quality and male fertility, the determination of seminal plasma cytokines provides a promising direction for research into the identification of factors connected with male infertility. The interleukins: IL-1α, -1ß, -2, -4, -6, -8, -10, -12p40, -12p70, -18, IFNγ, and GM-CSF, total oxidant (TOS) and antioxidant (TAS) status, were simultaneously examined in seminal plasmas and blood sera in terato- (n = 32), asthenoterato- (n = 33), and oligoasthenoteratozoospermic (n = 29) infertile men and in normozoospermic fertile men (n = 20). Our research shows different cytokine composition of the sera and seminal plasmas in all studied groups, along with much higher concentrations of seminal plasma GM-CSF, IFNγ, IL-1α, IL-4, IL-6, and IL-8 and lower IL-18 and TOS in the comparison to their sera levels. The seminal plasma concentrations of GM-CSF, IFNγ, IL-1α, -4, and -6 differ significantly between fertile and infertile as well as between teratozoospermic, asthenoteratozoospermic, and oligoasthenoteratozoospermic groups. The indication of the cause of different concentrations of cytokines in seminal plasmas of infertile men, and their associations with semen parameters and oxidative status, may be a promising direction for the search for new therapeutic targets that would directly affect the cells and tissues of male reproductive organs.

Asunto(s)

Antioxidantes , Biomarcadores , Citocinas , Infertilidad Masculina , Semen , Humanos , Masculino , Semen/metabolismo , Citocinas/metabolismo , Citocinas/sangre , Antioxidantes/metabolismo , Adulto , Biomarcadores/metabolismo , Biomarcadores/sangre , Infertilidad Masculina/metabolismo , Infertilidad Masculina/sangre , Estrés Oxidativo , Análisis de Semen/métodos , Espermatozoides/metabolismo , Astenozoospermia/metabolismo

RESUMEN

Oligoasthenozoospermia (OAS) is one of the most common types of male infertility, which, however, still lacks effective treatment. An increasing number of studies have shown the potential therapeutic value of omega-3 polyunsaturated fatty acid (ω-3 PUFA) in the treatment of OAS. This article presents an overview of the studies on the effects of ω-3 PUFA on fatty acid composition and metabolism, inflammatory response, and oxidative stress in OAS, hoping to provide some new ideas for the treatment of the disease.

Asunto(s)

Ácidos Grasos Omega-3 , Oligospermia , Humanos , Ácidos Grasos Omega-3/uso terapéutico , Masculino , Oligospermia/tratamiento farmacológico , Astenozoospermia/tratamiento farmacológico , Estrés Oxidativo/efectos de los fármacos

RESUMEN

PURPOSE: To find the machine learning (ML) method that has the highest accuracy in predicting the semen quality of men based on basic questionnaire data about lifestyle behavior. METHODS: The medical records of men whose semen was analyzed for any reason were collected. Those who had data about their lifestyle behaviors were included in the study. All semen analyses of the men included were evaluated according to the WHO 2021 guideline. All semen analyses were categorized as normozoospermia, oligozoospermia, teratozoospermia, and asthenozoospermia. The Extra Trees Classifier, Average (AVG) Blender, Light Gradient Boosting Machine (LGBM) Classifier, eXtreme Gradient Boosting (XGB) Classifier, Logistic Regression, and Random Forest Classifier techniques were used as ML algorithms. RESULTS: Seven hundred thirty-four men who met the inclusion criteria and had data about lifestyle behavior were included in the study. 356 men (48.5%) had abnormal semen results, 204 (27.7%) showed the presence of oligozoospermia, 193 (26.2%) asthenozoospermia, and 265 (36.1%) teratozoospermia according to the WHO 2021. The AVG Blender model had the highest accuracy and AUC for predicting normozoospermia and teratozoospermia. The Extra Trees Classifier and Random Forest Classifier models achieved the best performance for predicting oligozoospermia and asthenozoospermia, respectively. CONCLUSION: The ML models have the potential to predict semen quality based on lifestyles.

Asunto(s)

Estilo de Vida , Aprendizaje Automático , Análisis de Semen , Masculino , Humanos , Análisis de Semen/métodos , Adulto , Oligospermia/diagnóstico , Astenozoospermia/diagnóstico , Teratozoospermia/diagnóstico , Persona de Mediana Edad , Infertilidad Masculina/diagnóstico

RESUMEN

OBJECTIVE: To explore the potential impact of lipid metabolism-related single nucleotide polymorphisms (SNP) on semen quality in men. METHODS: We selected 284 semen samples from Xingtai Infertility Hospital and Hebei Human Sperm Bank collected between February and October 2023, 33 from oligozoospermia (OS), 97 from asthenozoospermia (AS) and 54 from oligoasthenozoospermia (OAS) patients and the other 100 from normal men. We performed computer-assisted semen analysis (CASA) of the samples, extracted blood DNA and, using the MassARRAY System, genotyped the target genes, determined the genotypes of 13 SNPs and compared their distribution, their correlation with BMI and semen quality in different groups. RESULTS: The mutant homozygous (TT) genotype of the FADS2 rs2727270 gene seemed to be a risk factor for AS (OR = 4.420, P= 0.047), while the APOA2 rs5082-A allele and MC4R rs17782313 heterozygous (TC) genotype important protective factors for OS (OR = 0.422 and 0.389; P= 0.045 and 0.043, respectively). A significantly higher sperm concentration was found associated with the MC4R rs17782313 heterozygous (TC) genotype than with the homozygous (CC) genotype. Stratification analysis showed that the protective effect of the TC genotype was decreased with increased BMI and remained with the interaction of the rs5082 and rs17782313 genotypes. CONCLUSION: FADS2 rs2727270, APOA2 rs5082 and MC4R rs17782313 were significantly correlated with the risk of abnormal semen parameters.

Asunto(s)

Genotipo , Metabolismo de los Lípidos , Polimorfismo de Nucleótido Simple , Análisis de Semen , Humanos , Masculino , Metabolismo de los Lípidos/genética , Astenozoospermia/genética , Ácido Graso Desaturasas/genética , Oligospermia/genética , Infertilidad Masculina/genética , Alelos , Adulto , Recuento de Espermatozoides , Factores de Riesgo , Espermatozoides/metabolismo

RESUMEN

Necrozoospermia is a special type of asthenospermia, in which mass sperm death is commonly seen, with an incidence rate of 0.2%－0.4%. Studies on necrospermia are rarely reported. Its etiology is complicated, and its diagnosis and treatment are very difficult. This article focuses on the main etiological factors, pathophysiological mechanism, diagnostic methods and treatment strategies of necrospermia, aiming to provide some reference for andrologists and reproduction physicians, as well as a theoretical guidance for intracytoplasmic sperm injection (ICSI) in the treatment of the patients with necrospermia.

Asunto(s)

Inyecciones de Esperma Intracitoplasmáticas , Humanos , Masculino , Inyecciones de Esperma Intracitoplasmáticas/métodos , Infertilidad Masculina/diagnóstico , Infertilidad Masculina/etiología , Infertilidad Masculina/terapia , Espermatozoides , Astenozoospermia/diagnóstico , Astenozoospermia/terapia

RESUMEN

OBJECTIVE: To observe the effect and safety of the traditional Chinese medicine (TCM) Linggui Formula (LGF) in the treatment of asthenospermia with kidney deficiency and blood stasis. METHODS: This randomized controlled trial included 90 cases of asthenospermia with kidney deficiency and blood stasis treated in our hospital from September 2022 to September 2023, 45 by oral medication with LGF (the trial group) and the other 45 with oral levocarnitine solution (the control group), all for 12 weeks. We followed up the patients for 12 weeks, recorded the semen parameters, TCM syndrome scores, sexual hormone levels, pregnancy rates, and DNA fragmentation index (DFI) of the patients, and compared them between the two groups before and after treatment. RESULTS: Totally, 82 of the patients completed the study, 42 in the trial and 40 in the control group. After treatment, the patients in the trial group showed significant increases in the percentage of progressively motile sperm (PMS) (from ï¼»19.25±3.08ï¼½% to ï¼»38.57±4.99ï¼½%, P< 0.05), total sperm motility (from ï¼»32.29±3.64ï¼½% to ï¼»46.50±4.77ï¼½%, P< 0.05) and sperm concentration (from ï¼»83.9±37.2ï¼½ to ï¼»95.1±34.9ï¼½× 106/ml ï¼½, P< 0.05), and so did the controls in PMS (from ï¼»19.75±4.28ï¼½% to ï¼»34.46±5.07ï¼½%, P< 0.05), total sperm motility (from ï¼»33.02±4.93ï¼½% to ï¼»43.11±4.72ï¼½%, P< 0.05) and sperm concentration (from ï¼»85.2±39.7ï¼½ to ï¼»88.1±35.2ï¼½ × 106/ml , P< 0.05), all even more significant in the trial than in the control group (P< 0.05). No statistically significant difference was observed in the semen volume either in the trial (from ï¼»3.38±0.38ï¼½ to ï¼»3.24±0.45ï¼½ ml, P> 0.05) or in the control group (from ï¼»3.46±0.52ï¼½ to ï¼»3.30±0.37ï¼½ ml, P> 0.05), or between the trial and control groups (P> 0.05), or in the sexual hormone levels, pregnancy rates, and sperm DFI between the two groups before and after treatment (P> 0.05). Both groups of patients had good safety profiles without serious adverse reactions. CONCLUSION: Linggui Formula can improve the percentage of PMS in asthenospermia patients with kidney deficiency and blood stasis, potentially enhancing pregnancy rates and with a good safety.

Asunto(s)

Astenozoospermia , Medicamentos Herbarios Chinos , Motilidad Espermática , Humanos , Masculino , Medicamentos Herbarios Chinos/uso terapéutico , Medicamentos Herbarios Chinos/administración & dosificación , Astenozoospermia/tratamiento farmacológico , Motilidad Espermática/efectos de los fármacos , Adulto , Medicina Tradicional China/métodos , Índice de Embarazo , Enfermedades Renales/tratamiento farmacológico , Enfermedades Renales/complicaciones , Embarazo , Fragmentación del ADN

RESUMEN

ABSTRACT: The cause of asthenozoospermia (AZS) is not well understood because of its complexity and heterogeneity. Although some gene mutations have been identified as contributing factors, they are only responsible for a small number of cases. Radial spokes (RSs) are critical for adenosine triphosphate-driven flagellar beating and axoneme stability, which is essential for flagellum motility. In this study, we found novel compound heterozygous mutations in leucine-rich repeat-containing protein 23 ( LRRC23 ; c.1018C>T: p.Q340X and c.881_897 Del: p.R295Gfs*32) in a proband from a nonconsanguineous family with AZS and male infertility. Diff-Quik staining and scanning electron microscopy revealed no abnormal sperm morphology. Western blotting and immunofluorescence staining showed that these mutations suppressed LRRC23 expression in sperm flagella. Additionally, transmission electron microscopy showed the absence of RS3 in sperm flagella, which disrupts stability of the radial spoke complex and impairs motility. Following in vitro fertilization and embryo transfer, the proband's spouse achieved successful pregnancy and delivered a healthy baby. In conclusion, our study indicates that two novel mutations in LRRC23 are associated with AZS, but successful fertility outcomes can be achieved by in vitro fertilization-embryo transfer techniques.

Asunto(s)

Astenozoospermia , Mutación , Humanos , Masculino , Astenozoospermia/genética , Adulto , Linaje , Cola del Espermatozoide/patología , Cola del Espermatozoide/ultraestructura , Cola del Espermatozoide/metabolismo , Femenino , Motilidad Espermática/genética , Embarazo

RESUMEN

Inner dynein arms (IDAs) are formed from a protein complex that is essential for appropriate flagellar bending and beating. IDA defects have previously been linked to the incidence of asthenozoospermia (AZS) and male infertility. The testes-enriched ZMYND12 protein is homologous with an IDA component identified in Chlamydomonas. ZMYND12 deficiency has previously been tied to infertility in males, yet the underlying mechanism remains uncertain. Here, a CRISPR/Cas9 approach was employed to generate Zmynd12 knockout (Zmynd12-/-) mice. These Zmynd12-/- mice exhibited significant male subfertility, reduced sperm motile velocity, and impaired capacitation. Through a combination of co-immunoprecipitation and mass spectrometry, ZMYND12 was found to interact with TTC29 and PRKACA. Decreases in the levels of PRKACA were evident in the sperm of these Zmynd12-/- mice, suggesting that this change may account for the observed drop in male fertility. Moreover, in a cohort of patients with AZS, one patient carrying a ZMYND12 variant was identified, expanding the known AZS-related variant spectrum. Together, these findings demonstrate that ZMYND12 is essential for flagellar beating, capacitation, and male fertility.

Asunto(s)

Infertilidad Masculina , Ratones Noqueados , Motilidad Espermática , Animales , Humanos , Masculino , Ratones , Astenozoospermia/genética , Astenozoospermia/metabolismo , Astenozoospermia/patología , Sistemas CRISPR-Cas , Dineínas/metabolismo , Dineínas/genética , Infertilidad Masculina/genética , Infertilidad Masculina/metabolismo , Infertilidad Masculina/patología , Ratones Endogámicos C57BL , Capacitación Espermática/genética , Motilidad Espermática/genética , Espermatozoides/metabolismo , Contactina 2/genética , Contactina 2/metabolismo

RESUMEN

TTC12 is a cytoplasmic and centromere-localized protein that plays a role in the proper assembly of dynein arm complexes in motile cilia in both respiratory cells and sperm flagella. This finding underscores its significance in cellular motility and function. However, the wide role of TTC12 in human spermatogenesis-associated primary ciliary dyskinesia (PCD) still needs to be elucidated. Whole-exome sequencing (WES) and Sanger sequencing were performed to identify potentially pathogenic variants causing PCD and multiple morphological abnormalities of sperm flagella (MMAF) in an infertile Pakistani man. Diagnostic imaging techniques were used for PCD screening in the patient. Real-time polymerase chain reaction (RT‒PCR) was performed to detect the effect of mutations on the mRNA abundance of the affected genes. Papanicolaou staining and scanning electron microscopy (SEM) were carried out to examine sperm morphology. Transmission electron microscopy (TEM) was performed to examine the ultrastructure of the sperm flagella, and the results were confirmed by immunofluorescence staining. Using WES and Sanger sequencing, a novel homozygous missense variant (c.C1069T; p.Arg357Trp) in TTC12 was identified in a patient from a consanguineous family. A computed tomography scan of the paranasal sinuses confirmed the symptoms of the PCD. RT-PCR showed a decrease in TTC12 mRNA in the patient's sperm sample. Papanicolaou staining, SEM, and TEM analysis revealed a significant change in shape and a disorganized axonemal structure in the sperm flagella of the patient. Immunostaining assays revealed that TTC12 is distributed throughout the flagella and is predominantly concentrated in the midpiece in normal spermatozoa. In contrast, spermatozoa from patient deficient in TTC12 showed minimal staining intensity for TTC12 or DNAH17 (outer dynein arms components). This could lead to MMAF and result in male infertility. This novel TTC12 variant not only illuminates the underlying genetic causes of male infertility but also paves the way for potential treatments targeting these genetic factors. This study represents a significant advancement in understanding the genetic basis of PCD-related infertility.

Asunto(s)

Homocigoto , Infertilidad Masculina , Mutación Missense , Cola del Espermatozoide , Humanos , Masculino , Mutación Missense/genética , Pakistán , Infertilidad Masculina/genética , Infertilidad Masculina/patología , Cola del Espermatozoide/patología , Cola del Espermatozoide/ultraestructura , Cola del Espermatozoide/metabolismo , Adulto , Linaje , Astenozoospermia/genética , Astenozoospermia/patología , Trastornos de la Motilidad Ciliar/genética , Trastornos de la Motilidad Ciliar/patología , Secuenciación del Exoma , Oligospermia/genética , Oligospermia/patología , Síndrome de Kartagener/genética , Síndrome de Kartagener/patología

RESUMEN

OBJECTIVE: To explore the expressions of zinc homeostasis-related proteins, G protein-coupled receptor 39 (GPR39) and ANO1 mRNA in the sperm of patients with asthenozoospermia (AS), and analyze their correlation with sperm motility. METHODS: We collected semen samples from 82 male subjects with PR+NP < 40%, PR < 32% and sperm concentration > 15×106/ml (the AS group, n = 40) or PR+NP ≥ 40%, PR ≥ 32% and sperm concentration > 15×106/ml (the normal control group, n = 42). We analyzed the routine semen parameters and measured the zinc content in the seminal plasma using the computer-assisted sperm analysis system, detected the expressions of zinc transporters (ZIP13, ZIP8 and ZNT10), metallothioneins (MT1G, MT1 and MTF), GPR39, and calcium-dependent chloride channel protein (ANO1) in the sperm by real-time quantitative PCR (RT qPCR), examined free zinc distribution in the sperm by laser confocal microscopy, and determined the expressions of GPR39 and MT1 proteins in the sperm by immunofluorescence staining, followed by Spearman rank correlation analysis of their correlation with semen parameters. RESULTS: There was no statistically significant difference in the zinc concentration in the seminal plasma between the AS and normal control groups (P>0.05). Compared with the controls, the AS patients showed a significantly reduced free zinc level (P<0.05), relative expressions of MT1G, MTF, ZIP13, GPR39 and ANO1 mRNA (P<0.05), and that of the GPR39 protein in the AS group (P<0.05). No statistically significant differences were observed in the relative expression levels of ZIP8, ZNT10 and MT1 mRNA between the two groups (P>0.05). The relative expression levels of GPR39, ANO1, MT1G and MTF mRNA were positively correlated with sperm motility and the percentage of progressively motile sperm (P<0.05). CONCLUSION: The expressions of zinc homeostasis proteins (MT1G, MTF and ZIP13), GPR39 and ANO1 mRNA are downregulated in the sperm of asthenozoospermia patients, and positively correlated with sperm motility.

Asunto(s)

Anoctamina-1 , Astenozoospermia , Proteínas de Transporte de Catión , ARN Mensajero , Receptores Acoplados a Proteínas G , Motilidad Espermática , Espermatozoides , Zinc , Humanos , Masculino , Astenozoospermia/metabolismo , Astenozoospermia/genética , Anoctamina-1/metabolismo , Anoctamina-1/genética , Receptores Acoplados a Proteínas G/metabolismo , Receptores Acoplados a Proteínas G/genética , Zinc/metabolismo , Espermatozoides/metabolismo , Proteínas de Transporte de Catión/metabolismo , Proteínas de Transporte de Catión/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Metalotioneína/metabolismo , Metalotioneína/genética , Homeostasis , Adulto , Análisis de Semen , Relevancia Clínica , Proteínas de Neoplasias

RESUMEN

N-linked glycoproteins are rich in seminal plasma, playing essential roles in supporting sperm function and fertilization process. The alteration of seminal plasma glycans and its correspond glycoproteins may lead to sperm dysfunction and even infertility. In present study, an integrative analysis of glycoproteomic and proteomic was performed to investigate the changes of site-specific glycans and glycoptoteins in seminal plasma of asthenozoospermia. By large scale profiling and quantifying 5,018 intact N-glycopeptides in seminal plasma, we identified 92 intact N-glycopeptides from 34 glycoproteins changed in asthenozoospermia. Especially, fucosylated glycans containing lewis x, lewis y and core fucosylation were significantly up-regulated in asthenozoospermia compared to healthy donors. The up-regulation of fucosylated glycans in seminal plasma may interfere sperm surface compositions and regulation of immune response, which subsequently disrupts sperm function. Three differentiated expression of seminal vesicle-specific glycoproteins (fibronectin, seminogelin-2, and glycodelin) were also detected with fucosylation alteration in seminal plasma of asthenozoospermia. The interpretation of the altered site-specific glycan structures provides data for the diagnosis and etiology analysis of male infertility, as well as providing new insights into the potential therapeutic targets for male infertility.

Asunto(s)

Astenozoospermia , Fucosa , Semen , Humanos , Masculino , Astenozoospermia/metabolismo , Semen/metabolismo , Semen/química , Fucosa/metabolismo , Glicoproteínas/metabolismo , Proteómica , Adulto , Regulación hacia Arriba , Polisacáridos/metabolismo , Polisacáridos/química , Glicosilación , Glicopéptidos/metabolismo , Glicopéptidos/análisis

RESUMEN

Aims: Asthenozoospermia is the most common factor of male infertility, mainly caused by multiple morphological abnormalities of the sperm flagella (MMAF) and primary ciliary dyskinesia (PCD). Previous studies have shown that genetic factors may contribute to MMAF and PCD. The study aimed to identify novel potentially pathogenic gene mutations in a Chinese infertile man with MMAF and PCD-like phenotypes. Methods: A Chinese infertile man with MMAF and PCD was enrolled in this study. Whole exome sequencing and Sanger sequencing were performed to identify potential causative genes and mutations. Results: A novel homozygous missense mutation (c.1450G>A; p.E484K) of CCDC40 was finally identified and Sanger sequencing confirmed that the patient carried the homozygous mutation, which was inherited from his parents. We reported the first homozygous missense CCDC40 mutation in infertile men with MMAF but had other milder PCD symptoms. Conclusion: Our findings not only broaden the disease-causing mutation spectrum of CCDC40 but also provide new insight into the correlation between CCDC40 mutations and MMAF.

Asunto(s)

Pueblo Asiatico , Homocigoto , Infertilidad Masculina , Mutación Missense , Fenotipo , Cola del Espermatozoide , Humanos , Masculino , Infertilidad Masculina/genética , Mutación Missense/genética , Adulto , China , Pueblo Asiatico/genética , Cola del Espermatozoide/metabolismo , Cola del Espermatozoide/patología , Trastornos de la Motilidad Ciliar/genética , Secuenciación del Exoma/métodos , Linaje , Mutación , Astenozoospermia/genética , Pueblos del Este de Asia

RESUMEN

Asthenospermia is a predominant cause of male infertility, and antioxidant supplements can be effective in treating asthenospermia. We demonstrate the antioxidant potential of traditional Chinese medicine, the Yishenhuoxue (YSHX) formula, in treating polyglycosides of Tripterygium wilfordii (GTW)-induced asthenospermia in rats. Fifty male rats were randomly divided into the normal, model, and treatment groups. HE staining was used to evaluate the improvement of spermatogenic function of rats, and TBA reaction, qRT-PCR, Western Blot and other methods were used to determine the changes of oxidative stress indicators and to evaluate the improvement of antioxidant capacity of rats by YSHX. Comparison with the model group showed significant improvement in pathological damage caused by GTW to seminiferous tubules. MDA and NO content in rat testes decreased, especially in middle- and high-dosage groups. No significant changes were observed in SOD and CAT activity or mRNA expression. GSH-Px activity and GSH mRNA expression were significantly higher in the low-dosage group than in the model group. Compared to the model group, GR activity was significantly lower in the middle and high dosage groups, while the mRNA expression was higher. The PKC-beta level increased, while p-ERK1/2, NF-κB, and the ratio of p-ERK1/2*(ERK1/2)-1 decreased significantly in the treatment groups. Therefore, YSHX can alleviate GTW-induced testicular damage, enhance GSH-Px activity, regulate GSH redox cycling, and mitigate oxidative stress injury. Furthermore, YSHX can promote PKC-beta expression and inhibit the phosphorylation of ERK1/2 and NF-κB. Using YSHX may be an effective way to increase sperm motility via the PKC-ERK1/2-NF-ĸB axis.

Asunto(s)

Antioxidantes , Astenozoospermia , Medicamentos Herbarios Chinos , Estrés Oxidativo , Ratas Sprague-Dawley , Animales , Masculino , Antioxidantes/farmacología , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéutico , Astenozoospermia/tratamiento farmacológico , Astenozoospermia/metabolismo , Estrés Oxidativo/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Ratas , FN-kappa B/metabolismo , Testículo/efectos de los fármacos , Testículo/metabolismo , Tripterygium/química , Malondialdehído/metabolismo , Superóxido Dismutasa/metabolismo

RESUMEN

Background: The involvement of ATP and cAMP in sperm function has been extensively documented, but the understanding of the role of adenosine and adenosine receptors remains incomplete. This study aimed to examine the presence of adenosine A2A receptor (A2AR) and study the functional role of A2AR in human sperm. Methods: The presence and localization of A2AR in human sperm were examined by western blotting and immunofluorescence assays. The functional role of A2AR in sperm was assessed by incubating human sperm with an A2AR agonist (regadenoson) and an A2AR antagonist (SCH58261). The sperm level of A2AR was examined by western blotting in normozoospermic and asthenozoospermic men to evaluate the association of A2AR with sperm motility and in vitro fertilization (IVF) outcomes. Results: A2AR with a molecular weight of 43 kDa was detected in the tail of human sperm. SCH58261 decreased the motility, penetration ability, intracellular Ca2+ concentration, and CatSper current of human sperm. Although regadenoson did not affect these sperm parameters, it alleviated the adverse effects of SCH58261 on these parameters. In addition, the mean level of A2AR in sperm from asthenozoospermic men was lower than that in sperm from normozoospermic men. The sperm level of A2AR was positively correlated with progressive motility. Furthermore, the fertilization rate during IVF was lower in men with decreased sperm level of A2AR than in men with normal sperm level of A2AR. Conclusions: These results indicate that A2AR is important for human sperm motility and is associated with IVF outcome.

Asunto(s)

Fertilización In Vitro , Receptor de Adenosina A2A , Motilidad Espermática , Espermatozoides , Humanos , Masculino , Motilidad Espermática/efectos de los fármacos , Receptor de Adenosina A2A/metabolismo , Espermatozoides/metabolismo , Espermatozoides/efectos de los fármacos , Fertilización In Vitro/métodos , Adulto , Astenozoospermia/metabolismo , Femenino , Pirazoles/farmacología , Agonistas del Receptor de Adenosina A2/farmacología , Antagonistas del Receptor de Adenosina A2/farmacología , Pirimidinas/farmacología , Triazoles/farmacología

RESUMEN

Background: Asthenozoospermia, a type of male infertility, is primarily caused by dysfunctional sperm mitochondria. Despite previous bioinformatics analysis identifying potential key lncRNAs, miRNAs, hub genes, and pathways associated with asthenospermia, there is still a need to explore additional molecular mechanisms and potential biomarkers for this condition. Methods: We integrated data from Gene Expression Omnibus (GEO) (GSE22331, GSE34514, and GSE160749) and performed bioinformatics analysis to identify differentially expressed genes (DEGs) between normozoospermia and asthenozoospermia. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were conducted to gain insights into biological processes and signaling pathways. Weighted Gene Co-expression Network Analysis (WGCNA) identified gene modules associated with asthenozoospermia. Expression levels of key genes were assessed using datasets and experimental data. Gene Set Enrichment Analysis (GSEA) and correlation analysis identified pathways associated with the hub gene and explore the relationship between the ZNF764 and COQ9 and mitochondrial autophagy-related genes. Competitive endogenous RNA (ceRNA) networks were constructed, and in vitro experiments using exosome samples were conducted to validate this finding. Results: COQ9 was identified as a marker gene in asthenozoospermia, involved in autophagy, ATP-dependent chromatin remodeling, endocytosis, and cell cycle, etc. The ceRNA regulatory network (LINC00893/miR-125a-5p/COQ9) was constructed, and PCR demonstrated that LINC00893 and COQ9 were downregulated in asthenozoospermia, while miR-125a-5p and m6A methylation level of LINC00893 were upregulated in asthenozoospermia compared to normozoospermic individuals. Conclusion: The ceRNA regulatory network (LINC00893/miR-125a-5p/COQ9) likely plays a crucial role in the mechanism of asthenozoospermia. However, further functional experiments are needed to fully understand its significance.

Asunto(s)

Astenozoospermia , Biomarcadores , Biología Computacional , Redes Reguladoras de Genes , Humanos , Masculino , Astenozoospermia/genética , Astenozoospermia/metabolismo , Biología Computacional/métodos , Biomarcadores/metabolismo , Perfilación de la Expresión Génica , MicroARNs/genética , MicroARNs/metabolismo , Ontología de Genes , Transducción de Señal/genética , Espermatozoides/metabolismo