Prognosis Of Male Infertility Research Articles

DNAH3 deficiency causes flagellar inner dynein arm loss and male infertility in humans and mice

Axonemal protein complexes, including the outer and inner dynein arms (ODA/IDA), are highly ordered structures of the sperm flagella that drive sperm motility. Deficiencies in several axonemal proteins have been associated with male infertility, which is characterized by asthenozoospermia or asthenoteratozoospermia. Dynein axonemal heavy chain 3 (DNAH3) resides in the IDA and is highly expressed in the testis. However, the relationship between DNAH3 and male infertility is still unclear. Herein, we identified biallelic variants of DNAH3 in four unrelated Han Chinese infertile men with asthenoteratozoospermia through whole-exome sequencing (WES). These variants contributed to deficient DNAH3 expression in the patients’ sperm flagella. Importantly, the patients represented the anomalous sperm flagellar morphology, and the flagellar ultrastructure was severely disrupted. Intriguingly, Dnah3 knockout (KO) male mice were also infertile, especially showing the severe reduction in sperm movement with the abnormal IDA and mitochondrion structure. Mechanically, nonfunctional DNAH3 expression resulted in decreased expression of IDA-associated proteins in the spermatozoa flagella of patients and KO mice, including DNAH1, DNAH6, and DNALI1, the deletion of which has been involved in disruption of sperm motility. Moreover, the infertility of patients with DNAH3 variants and Dnah3 KO mice could be rescued by intracytoplasmic sperm injection (ICSI) treatment. Our findings indicated that DNAH3 is a novel pathogenic gene for asthenoteratozoospermia and may further contribute to the diagnosis, genetic counseling, and prognosis of male infertility.

DNAH3 deficiency causes flagellar inner dynein arm loss and male infertility in humans and mice.

Axonemal protein complexes, including the outer and inner dynein arms (ODA/IDA), are highly ordered structures of the sperm flagella that drive sperm motility. Deficiencies in several axonemal proteins have been associated with male infertility, which is characterized by asthenozoospermia or asthenoteratozoospermia. Dynein axonemal heavy chain 3 (DNAH3) resides in the IDA and is highly expressed in the testis. However, the relationship between DNAH3 and male infertility is still unclear. Herein, we identified biallelic variants of DNAH3 in four unrelated Han Chinese infertile men with asthenoteratozoospermia through whole-exome sequencing (WES). These variants contributed to deficient DNAH3 expression in the patients' sperm flagella. Importantly, the patients represented the anomalous sperm flagellar morphology, and the flagellar ultrastructure was severely disrupted. Intriguingly, Dnah3 knockout (KO) male mice were also infertile, especially showing the severe reduction in sperm movement with the abnormal IDA and mitochondrion structure. Mechanically, nonfunctional DNAH3 expression resulted in decreased expression of IDA-associated proteins in the spermatozoa flagella of patients and KO mice, including DNAH1, DNAH6, and DNALI1, the deletion of which has been involved in disruption of sperm motility. Moreover, the infertility of patients with DNAH3 variants and Dnah3 KO mice could be rescued by intracytoplasmic sperm injection (ICSI) treatment. Our findings indicated that DNAH3 is a novel pathogenic gene for asthenoteratozoospermia and may further contribute to the diagnosis, genetic counseling, and prognosis of male infertility.

Novel biallelic mutations in TTC29 cause asthenoteratospermia and male infertility.

Multiple morphological abnormalities of the sperm flagella (MMAF), which is characterized as asthenoteratospermia involving absent, short, bent, coiled, and/or irregular-caliber flagella, is a rare recessive inherited disorder associated with male infertility. To date, genetic causes of MMAF cases are not fully explored. Whole-exome sequencing was conducted to identify pathogenic variants in a patient with MMAF. The functional effect of the identified mutations was investigated by immunofluorescence staining and western blotting. Intracytoplasmic sperm injection was used to assist fertilization for the patient with MMAF. We identified novel biallelic mutations, a splicing variant NC_000004.12:g.146937593C>T (c.254+1G>A), and a nonsense mutation NM_001300761.4:c.1185C>G (NP_001287690.1:p.Tyr395*), in TTC29 from an infertile patient. In addition to the typical MMAF phenotype, the patient also presented aberrant morphology of sperm heads. Further functional experiments confirmed the absence of TTC29 expression in the spermatozoa. We also explored the specific expression pattern of TTC29 in human and mouse spermatogenesis. The outcome of intracytoplasmic sperm injection in the patient was unsuccessful, while additional female risk factors should not be excluded. Our study revealed the novel biallelic mutations in TTC29 in a MMAF patient, which findings expand the mutational spectrum of TTC29 and further contribute to the diagnosis, genetic counseling, and prognosis of male infertility.

A new method for evaluating the quality of single sperm by detecting reactive oxygen species

Sperm damage caused by reactive oxygen species (ROS) is one of the main causes of male infertility. Therefore, the level of ROS in sperm is an important indicator for the diagnosis and prognosis of male infertility. Herein, we constructed a single sperm ROS detection method (SSRDM) with an optical micro-probe fabricated via focused ion beam process. The micro-probe is used to separately excite fluorescence in the sperm and the area around the sperm after ROS staining, and the difference in fluorescence values can reflect the level of ROS in the sperm. We collected 102 semen samples and 72 of them were divided into asthenozoospermia and non-asthenozoospermia groups. SSRDM and flow cytometry were used to detect the ROS levels of the two groups. The results of SSRDM showed that the ROS levels of asthenozoospermia group were higher than that of non-asthenozoospermia group (P = 0.002), while the results of flow cytometry indicated no difference (P = 0.152). In terms of ROS levels, compared with flow cytometry, SSRDM has a stronger ability to distinguish between those two groups, providing a reliable basis for assessment of sperm quality. Another 30 semen samples were used to investigate temperature and temporal variability of SSRDM to ensure the stability and accuracy of this method. Overall, we have developed a method that can quantitatively detect fluorescent substances in sperm at the single-cell level supplying evidence for diagnosis and prognosis of male infertility.

Oxidation of Sperm DNA and Male Infertility.

One important reason for male infertility is oxidative stress and its destructive effects on sperm structures and functions. The particular composition of the sperm membrane, rich in polyunsaturated fatty acids, and the easy access of sperm DNA to oxidative damage due to sperm cell specific cytologic and metabolic features (no cytoplasm left and cells unable to mount stress responses) make it the cell type in metazoans most susceptible to oxidative damage. In particular, oxidative damage to the spermatozoa genome is an important issue and a cause of male infertility, usually associated with single- or double-strand paternal DNA breaks. Various methods of detecting sperm DNA fragmentation have become important diagnostic tools in the prognosis of male infertility and such assays are available in research laboratories and andrology clinics. However, to date, there is not a clear consensus in the community as to their respective prognostic value. Nevertheless, it is important to understand that the effects of oxidative stress on the sperm genome go well beyond DNA fragmentation alone. Oxidation of paternal DNA bases, particularly guanine and adenosine residues, the most sensitive residues to oxidative alteration, is the starting point for DNA damage in spermatozoa but is also a danger for the integrity of the embryo genetic material independently of sperm DNA fragmentation. Due to the lack of a spermatozoa DNA repair system and, if the egg is unable to correct the sperm oxidized bases, the risk of de novo mutation transmission to the embryo exists. These will be carried on to every cell of the future individual and its progeny. Thus, in addition to affecting the viability of the pregnancy itself, oxidation of the DNA bases in sperm could be associated with the development of conditions in young and future adults. Despite these important issues, sperm DNA base oxidation has not attracted much interest among clinicians due to the lack of simple, reliable, rapid and consensual methods of assessing this type of damage to the paternal genome. In addition to these technical issues, another reason explaining why the measurement of sperm DNA oxidation is not included in male fertility is likely to be due to the lack of strong evidence for its role in pregnancy outcome. It is, however, becoming clear that the assessment of DNA base oxidation could improve the efficiency of assisted reproductive technologies and provide important information on embryonic developmental failures and pathologies encountered in the offspring. The objective of this work is to review relevant research that has been carried out in the field of sperm DNA base oxidation and its associated genetic and epigenetic consequences.

Characterization of human prostasomes protein Clusterin (macromolecule) – a novel biomarker for male infertility diagnosis and prognosis

Prostasomes, a secretory particle from prostate gland in human seminal fluid plays a role in enhancing the fertility and its absence or less presence will lead to male infertility. Very few fertility associated proteins were detected in prostasomes. In order to isolate the prostasomes from ejaculated semen, the characterization was done using biochemical, molecular and in silico methods. The objective of current work on prostasomes is to identify a novel protein biomarker for the diagnosis and prognosis of male infertility. Semen samples were collected, primary semen analysis was done and prostasomes were isolated from ejaculated semen of fertile and infertile categories. Biochemical parameters like protein content, total antioxidant capacity, cholesterol content were evaluated in prostasomes. The critical expressed protein was identified by using SDS page and MALDI TOF techniques. Isolated particle from the semen samples was confirmed as prostasomes from SEM analysis and particle size analyzer. In MALDI results, the maximum hit was obtained against Clusterin that been reported to be involved in capacitation and motility of sperm. Structural modeling and molecular dynamics were carried out on Clusterin and elucidate the structural insights in the dynamic system. Overall, our study reported the novel biomarker Clusterin in prostasomes for diagnosis and prognosis of male infertility. Communicated by Ramaswamy H. Sarma

Sperm telomere length: Diagnostic and prognostic biomarker in male infertility (Review)

Telomere length is an important biomarker in a number of diseases, including male infertility. A decreased telomere length has been reported in several diseases and telomere shortening may occur due to aging, decreased telomerase activity, oxidative stress or cell division. In recent years, several studies have indicated that males with infertility have a shorter sperm telomere length than fertile males. Sperm telomere shortening is associated with male infertility through several mechanisms, including the apoptosis of spermatozoa, decreased motility, low sperm count, incorrect chromosomal pairing and movement during meiosis, and failed fertilization. The aim of this review was to compile current findings on sperm telomere length and discuss findings to compare sperm from infertile males with that of fertile males. Several studies reported shortening of sperm telomeres associated with infertility. Thus, sperm telomere length can be used as biomarker for the diagnosis and prognosis of male infertility, since fertile males have longer telomeres and the length decreases with age. However, there is no specific telomere length that is set as standard/recommended length.

Exosomal microRNAs in seminal plasma are markers of the origin of azoospermia and can predict the presence of sperm in testicular tissue.

STUDY QUESTIONAre exosomal microRNAs (miRNAs) in seminal plasma (SP) useful as markers of the origin of azoospermia and the presence of sperm in the testis?SUMMARY ANSWEROur study demonstrated the potential of several miRNAs contained in small extracellular vesicles (sEVs) of seminal fluid as sensitive and specific biomarkers for selecting those azoospermic individuals with real chances of obtaining spermatozoa from the testicular biopsy.WHAT IS KNOWN ALREADYThere are no precise non-invasive diagnostic methods for classifying the origin of the sperm defects in semen and the spermatogenic reserve of the testis in those infertile men with a total absence of sperm in the ejaculate (azoospermia). The diagnosis of such individuals is often based on the practice of biopsies. In this context it is reasonable to study the presence of organ-specific markers in human semen that contains fluid from the testis and the male reproductive glands, which could help in the diagnosis and prognosis of male infertility. Additionally, seminal fluid contains high concentrations of sEVs that are morphologically and molecularly consistent with exosomes, which originate from multiple cellular sources in the male reproductive tract.STUDY DESIGN, SIZE, DURATIONA case and control prospective study was performed. This study compares the miRNA content of exosomes in semen samples obtained from nine normozoospermic fertile individuals (control group), 14 infertile men diagnosed with azoospermia due to spermatogenic failure, and 13 individuals with obstructive azoospermia and conserved spermatogenesis. Additionally, three severe oligozoospermic individuals (<5 × 106 sperm/ml) were included in the study.PARTICIPANTS/MATERIALS, SETTING, METHODSA differential high-throughput miRNA profiling analysis using miRNA quantitative PCR panels was performed in SP exosomes from azoospermic patients and fertile individuals.MAIN RESULTS AND THE ROLE OF CHANCEA total of 623 miRNAs were included in the miRNA profiling stage of the study. A total of 397 miRNAs (63.7%) were consistently detected in samples from all groups and statistically analysed, which revealed altered patterns of miRNA expression in infertile patients. We focused on the miRNAs that were differentially expressed between azoospermia as a result of an obstruction in the genital tract (i.e. having conserved spermatogenesis) and azoospermia caused by spermatogenic failure, and described, in a miRNA validation stage of the study, the expression values of one miRNA (miR-31-5p) in exosomes from semen as a predictive biomarker test for the origin of azoospermia with high sensitivity and specificity (>90%). The efficacy of the predictive test was even better when the blood FSH values were included in the analysis. Furthermore a model that included miR-539-5p and miR-941 expression values is also described as being useful for predicting the presence of residual spermatogenesis in individuals with severe spermatogenic disorders with diagnostic accuracy.LIMITATIONS, REASONS FOR CAUTIONFurther studies, with an independent second population involving a larger number of samples, are needed to confirm our findings.WIDER IMPLICATIONS OF THE FINDINGSOur findings contribute to the search for the most valuable genetic markers that are potentially useful as tools for predicting the presence of testicular sperm in azoospermic individuals.STUDY FUNDING/COMPETING INTEREST(S)This work was financially supported by grants from the Fondo de Investigaciones Sanitarias/Fondo Europeo de Desarrollo Regional “Una manera de hacer Europa” (FIS/FEDER) [Grant number PI15/00153], the Generalitat de Catalunya [Grant number 2014SGR5412]. S.L. is sponsored by the Researchers Stabilization Program (ISCIII/Generalitat de Catalunya) from the Spanish National Health System [CES09/020].

Diagnosis and prognosis of male infertility in mammal: the focusing of tyrosine phosphorylation and phosphotyrosine proteins.

Male infertility refers to the inability of a man to achieve a pregnancy in a fertile female. In more than one-third of cases, infertility arises due to the male factor. Therefore, developing strategies for the diagnosis and prognosis of male infertility is critical. Simultaneously, a satisfactory model for the cellular mechanisms that regulate normal sperm function must be established. In this regard, tyrosine phosphorylation is one of the most common mechanisms through which several signal transduction pathways are adjusted in spermatozoa. It regulates the various aspects of sperm function, for example, motility, hyperactivation, capacitation, the acrosome reaction, fertilization, and beyond. Several recent large-scale studies have identified the proteins that are phosphorylated in spermatozoa to acquire fertilization competence. However, most of these studies are basal and have not presented an overall mechanism through which tyrosine phosphorylation regulates male infertility. In this review, we focus of this mechanism, discussing most of the tyrosine-phosphorylated proteins in spermatozoa that have been identified to date. We categorized tyrosine-phosphorylated proteins in spermatozoa that regulate male infertility using MedScan Reader (v5.0) and Pathway Studio (v9.0).

Lipid Peroxidation, Sperm DNA Fragmentation Total Antioxidant Capacity and Semen Quality in Male Infertility

Infertility is a major clinical problem, affecting people medically and psychosocially. Male factor plays a significant role in about 50% of infertile couples. Recent reports indicate that increasing male infertility could be due to genomic abnormalities. The etiology of sperm DNA damage is multi-factorial but compromised due to nuclear defects, protamine deficiency and oxidative stress. The present study was aimed to evaluate sperm DNA integrity and oxidative stress in infertile men. The study is prospective, comprises 96 infertile patients and 30 fertile controls. Sperm DNA integrity was assessed by flowcytometry. MDA and TAC were evaluated spectrophotometrically. The percentage of DNA Fragmentation Index and MDA were found to be significantly increased while TAC was significantly decreased in infertile men as compared to control. DFI and MDA were negatively correlated with TAC levels. Present study indicates significant increases in seminal MDA and sperm DNA damage in infertile men. Seminal MDA was significantly correlated with sperm DNA damage, TAC and standard sperm parameters. The elevated levels of seminal OS observed in these infertile patients could be responsible for poor sperm quality and sperm DNA fragmentation. Hence evaluation of DFI, MDA and TAC can be used for diagnosis, prognosis of male infertility in addition to routine semen parameters to decide the treatment strategies.

Functional testicular evaluation using PET/CT with 18F-fluorodeoxyglucose

PET/CT using (18)F-FDG is a well-established diagnostic examination in oncology, cardiology and neurology. The clinical significance of nontumoral testicular uptake of FDG is unknown. Functional testicular imaging may have important clinical applications in the diagnosis and prognosis of male infertility. The aim of this study was to determine the andrological value of a FDG PET/CT in analysing testicular function, by correlating the PET/CT data with the sperm parameters. Retrospective analysis of FDG PET/CT in 20 consecutive cancer patients without testicular pathology in whom two semen samples had been obtained for analysis before any chemotherapy. FDG PET/CT parameters were the mean standardized uptake value (SUVmean), used for measuring the intensity of uptake, and the functional testicular volume (FV). For statistical analysis, a Spearman's rank correlation test and a Mann-Whitney test were used. Of 20 patients (mean age 22 years), 18 had provided two sperm samples for cryopreservation. Sperm concentration was above 20 × 10(6)/ml in 55% of the patients. The intensity of uptake and the FV were correlated with the total sperm count, the sperm concentration and motility (p < 0.05). The difference in SUVmean between the two testes showed an inverse correlation with sperm concentration (p = 0.036). Normospermic and oligospermic men had significant differences in: (1) mean SUVmean, (2) mean FV, and (3) the difference in intensity of uptake between the testes (p < 0.05). This is the first report on the andrological value of FDG PET/CT in analysing nontumoral testicular function. This pilot study showed a significant correlation between intensity of uptake of FDG and testicular FV with the main sperm parameters. PET/CT with FDG could become a useful new tool in assisted reproductive technologies and other andrological or urological applications.

Rôle de l'urologue dans la prise en charge de l'homme infertile à l'heure de l'injection intracytoplasmique de spermatozoı̈des

Although intracytoplasmic sperm injection (ICSI) revolutionized treatment and prognosis of male infertility, check-up (case history, clinical and paraclinical examinations) practised by urologist in infertile man keeps all its place. Varicocele, congenital or acquired seminal duct obstruction, urogenital tract infection, or ejaculation disorder must be sought, because these affections remain accessible to treatment.

Radiozinc Spermatogenesis Test: Follow-Up Semen Analysis

Semen analysis was performed on 8 men, every 2 months, for a period of 8 months, after the application of the radiozinc spermatogensis test. Semen was examined for volume, cell count, motility, and abnormal forms. Results showed no evidence of any changes in semen parameters. Student's test was used to compare the data before the test with the data after the test, and no significant differences were seen. The radiozinc spermatogensis test seems safe, from a radiologic point of view, does not have an adverse effect on semen, and unlike testicular biopsy, saves the patient from surgical exposure. The radiozinc spermatogensis test can be applied as an important adjuvant measure in the diagnosis and prognosis of male infertility.

The Diagnosis and Prognosis of Male Infertility: A Study of 44 Cases, with Special Reference to Sperm Morphology

The Diagnosis and Prognosis of Male Infertility: A Study of 44 Cases, with Special Reference to Sperm Morphology