Adult ; Gene Deletion ; Humans ; Infertility, Male/genetics* ; Male ; Membrane Proteins/genetics* ; Teratozoospermia/genetics* ; Translocation, Genetic

Objective: To investigate the association of a very common mutation of c.144delC in the aurora kinase C (AURKC) gene with idiopathic teratozoospermia in Chinese infertile men in Sichuan. METHODS: Using polymerase chain reaction (PCR) and next-generation sequencing, we analyzed the correlation between c.144delC polymorphism of the AURKC gene and male infertility in 98 idiopathic teratozoospermia patients in comparison with 162 normal fertile men. RESULTS: Neither c.144delC mutation nor other meaningful mutations were detected in the AURKC gene in the 98 idiopathic teratozoospermia patients or the 162 normal controls. CONCLUSIONS Teratozoospermia is not correlated with c.144delC mutation in the AURKC gene in the men of the Sichuan area. Therefore, large-scale genotyping of the AURKC gene may not be necessary clinically among Chinese patients with idiopathic teratozoospermia.
Aurora Kinase C ; genetics ; Humans ; Male ; Mutation ; genetics ; Polymorphism, Genetic ; Spermatozoa ; Teratozoospermia ; genetics

OBJECTIVE: To explore the genetic basis for 4 patients with globozoospermia. METHODS: Semen and blood samples were collected from the patients for the determination of sperm concentration, viability, survival rate, morphology and acrosome antigen CD46. Meanwhile, DNA was extracted for whole exome sequencing (WES), and candidate variants were validated by Sanger sequencing. RESULTS: All of the four patients were found to harbor variants of the DPY19L2 gene. Patients 1 ~ 3 had homozygous deletions of the DPY19L2 gene. Sanger sequencing confirmed that the DPY19L2 gene in patient 3 was disrupted at a recombination breakpoint area BP2, resulting in nonallelic homologous recombination and complete deletion of the DPY19L2 gene. Patients 2 and 3 respectively harbored novel homozygous deletions of exons 2 ~ 22 and exons 14 ~ 15. Patient 4 harbored heterozygous deletion of the DPY19L2 gene, in addition with a rare homozygous deletion of the 3' UTR region. CONCLUSION DPY19L2 gene variants probably underlay the globozoospermia in the four patients, which has fit an autosomal recessive pattern of inheritance and the characteristics of genomic diseases.
Male ; Humans ; Teratozoospermia/genetics* ; Homozygote ; Semen ; Sequence Deletion ; 3' Untranslated Regions ; Membrane Proteins

OBJECTIVE: To study the association of the level of advanced oxidation protein products (AOPPs) in seminal plasma with teratospermia and the outcome parameters of fertilization (IVF). METHODS: We conducted a cross-sectional study among 272 male patients receiving assisted reproduction treatment in the Center for Reproductive Medicine of our hospital between October, 2018 and March, 2019. The levels of seminal AOPPs and reactive oxygen species (ROS), demographic data, sperm parameters and IVF outcome parameters were analyzed for all the patients. According to the percentage of sperms with normal morphology, the patients were divided before IVF into teratozoospermia group and normal sperm morphology group, and those in teratozoospermia group were further divided into 3 subgroups with mild, moderate and severe teratozoospermia. The patients were also divided on the day oocyte retrieval into 2 groups with fertilizing rates lower (group Ⅰ) and higher (group Ⅱ) than the median rate. RESULTS: We found a significant negative correlation of seminal AOPP level before treatment with the percentage of normal sperm morphology (=0.003) and seminal ROS level (=0.013). The seminal levels of AOPPs (= 0.027) and ROS (=0.036) were significantly elevated in patients with teratospermia, and seminal AOPP level was significantly higher in severe teratospermia group than in mild (=0.019) and moderate (=0.015) teratospermia groups. The seminal levels of AOPPs (=0.003) and ROS (=0.017) on the day of oocyte retrieval were negatively correlated with the fertilization rate in IVF cycles, and the levels of AOPPs (=0.049) and ROS (=0.036) were significantly higher in group Ⅰ than in group Ⅱ. CONCLUSIONS An elevated level of seminal AOPPs may indicate an increased risk of severe teratospermia and a lower fertilization rate in IVF.
Advanced Oxidation Protein Products ; Cross-Sectional Studies ; Fertilization in Vitro ; Humans ; Male ; Semen ; Spermatozoa ; Teratozoospermia

Autophagy is involved in spermatogenesis by regulating germ cell maturation. This catabolic process increases with hyperthermic conditions to prevent the accumulation of damaged organelles. Cryptorchidism is associated with impairment of germ cell maturation revealed by the presence of immature forms of sperm cells in ejaculates. The aim of the present study was to evaluate the status of autophagy in sperm cells from cryptorchid patients. Semen samples of cryptorchid patients and normozoospermic controls were analyzed by immunocytochemistry and electron microscopy. Autophagy proteins, autophagy-related protein 9 (ATG9) and microtubule-associated protein, 1A/1B-light chain 3 (LC3) were localized by immunocytochemistry on the acrosome and on the equatorial segment of sperm cells. LC3 was also detected in the midpiece of cryptorchid sperm tail. Autophagy substrate p62 protein was present in the acrosome and in the postequatorial segment of sperm in control samples, but not in the cryptorchid ones. Transmission electron microscopy revealed double-membrane-limited autophagosomes in postequatorial part of spermatozoa head and midpiece in cryptorchid samples. Partly degraded mitochondria were frequently discerned in autophagic vacuoles. In conclusion, autophagy is increased in sperm cells from patients with cryptorchid history comparatively to control. Our work provides insights into the role of autophagy in the maturation and survival of human male gametes in pathological conditions. Thus, regulating autophagy could represent a potential way to improve sperm quality in cryptorchid men.
Adult ; Autophagy ; Case-Control Studies ; Cryptorchidism/pathology* ; Humans ; Male ; Microscopy, Electron, Transmission ; Spermatogenesis ; Spermatozoa/pathology* ; Teratozoospermia/pathology* ; Testis/pathology*

Thanks to tremendous advances in sequencing technologies and in particular to whole exome sequencing (WES), many genes have now been linked to severe sperm defects. A precise genetic diagnosis is obtained for a minority of patients and only for the most severe defects like azoospermia or macrozoospermia which is very often due to defects in the aurora kinase C (AURKC gene. Here, we studied a subject with a severe oligozoospermia and a phenotypic diagnosis of macrozoospermia. AURKC analysis did not reveal any deleterious variant. WES was then initiated which permitted to identify a homozygous loss of function variant in the zinc finger MYND-type containing 15 (ZMYND15 gene. ZMYND15 has been described to serve as a switch for haploid gene expression, and mice devoid of ZMYND15 were shown to be sterile due to nonobstructive azoospermia (NOA). In man, ZMYND15 has been associated with NOA and severe oligozoospermia. We confirm here that the presence of a bi-allelic ZMYND15 variant induces a severe oligozoospermia. In addition, we show that severe oligozoospermia can be associated macrozoospermia, and that a phenotypic misdiagnosis is possible, potentially delaying the genetic diagnosis. In conclusion, genetic defects in ZMYND15 can induce complete NOA or severe oligozoospermia associated with a very severe teratozoospermia. In our experience, severe oligozoospermia is often associated with severe teratozoospermia and can sometimes be misinterpreted as macrozoospermia or globozoospermia. In these instances, specific AURKC or dpy-19 like 2 (DPY19L2) diagnosis is usually negative and we recommend the direct use of a pan-genomic techniques such as WES.
Animals ; Azoospermia/genetics* ; Humans ; Infertility, Male/genetics* ; Male ; Membrane Proteins/genetics* ; Mice ; Mutation ; Oligospermia/genetics* ; Repressor Proteins/metabolism* ; Teratozoospermia/genetics*

OBJECTIVE: To explore the molecular basis for two brothers affected with globozoospermia. METHODS: Whole exome sequencing was carried out for both patients. Candidate variant was verified by Sanger sequencing and quantitative real-time PCR (qRT-PCR). RESULTS: Whole exome sequencing, Sanger sequencing and qRT-PCR verification revealed a heterozygous c.384dup (p.Glu129*) variant in the DPY19L2 gene in the two brothers and their mother. A large heterozygous deletion, spanning approximately 164.5 kb and encompassing the entire DPY19L2 gene, was detected on chromosome 12 of the two patients and their father. CONCLUSION The c.384dup (p.Glu129*) variant and deletion of the DPY19L2 gene probably underlie the pathogenesis of globozoospermia in the two patients, which was in keeping with the autosomal recessive inheritance of disease in this pedigree.
Gene Deletion ; Genetic Variation ; Humans ; Infertility, Male ; genetics ; Male ; Membrane Proteins ; genetics ; Pedigree ; Siblings ; Teratozoospermia ; genetics ; Whole Exome Sequencing

OBJECTIVETo explore genetic mutation and clinical treatment for a patient with globozoospermia.

METHODSHistomorphology of the sperms was studied by Wright-Giemsa staining and transmission electron microscopy. Potential mutation of the DPY19L2 gene was detected by PCR amplification and Sanger sequencing.

RESULTSWright-Giemsa staining showed that all spermatozoa from the patient were round-headed and lacked the acrosome, with the nuclei of sperm head stained in dark and full. Transmission electron microscopy revealed large round sperm heads, with an even layer of unit membrane surrounding the nuclei and dispersed cytoplasmic vacuoles but no acrosomal structure. The patient has harbored a homozygous deletion of the DPY19L2 gene. With intracytoplasmic sperm injection (ICSI) treatment, fertilization rate of the oocytes has reached 28.6%, which resulted in a successful pregnancy. A healthy male was born.

CONCLUSIONThe homozygous deletion of DPY19L2 probably underlies the globozoospermia in this case, for which ICSI has provided an effective treatment. However, there is still a risk of low oocyte fertilization rate or fertilization failure. Further studies are required.

Adult ; DNA Mutational Analysis ; Humans ; Infant, Newborn ; Male ; Membrane Proteins ; genetics ; Mutation ; Sperm Injections, Intracytoplasmic ; Teratozoospermia ; genetics

Objective: To investigate the routine semen parameters and sperm morphological indexes of the patients with partial globozoospermia (PGZ). METHODS: We included in this study 100 infertile males with PGZ and another 180 non-PGZ infertile men as controls. According to the proportion of round-headed sperm (RHS) in the semen, we classified the PGZ males into five subgroups: 25%－40%, 41%－55%, 56%－70%, 71%－85%, and 86%－99% RHS. We obtained sperm concentration, total sperm motility, the percentage of progressively motile sperm, teratozoospermia index (TZI), and sperm deformity index (SDI) from the subjects and compared them among different groups. RESULTS: Statistically significant differences were found between the PGZ patients and non-PGZ controls in total sperm motility (［35.76±24.88］% vs ［62.03±10.20］%, P<0.01), the percentage of progressively motile sperm (［26.11±20.39］% vs ［45.62±6.87］%, P<0.01), the percentage of morphologically normal sperm (［1.45±1.45］% vs ［5.98±2.21］%, P<0.01), and SDI (1.33±0.11 vs 1.27±0.57, P<0.01), but not in age (［29.82±4.90］ vs ［30.33±3.59 ］ yr, P>0.05), sperm concentration (［46.01±40.38］ vs ［54.00±25.15］ ×106/ml, P>0.05), or TZI (1.35±0.11 vs 1.34±0.54, P>0.05). There were also significant differences among the five PGZ subgroups in total sperm motility, progressive sperm motility, normal sperm morphology, TZI, and SDI (P<0.01), but not in age or sperm concentration (P>0.05). Morphologically, the sperm head changed from heterogeneous to homogeneous with the increased proportion of round-headed sperm. CONCLUSIONS Different proportions of round-headed sperm are closely related to routine semen parameters and sperm morphological index in PGZ patients, which can help clinicians choose the proper assisted reproductive technology and predict the rate of fertilization for infertile males.
Case-Control Studies ; Humans ; Infertility, Male ; pathology ; Male ; Semen Analysis ; Sperm Count ; Sperm Head ; pathology ; Sperm Motility ; Spermatozoa ; abnormalities ; Teratozoospermia ; pathology

Teratozoospermia is a rare disease associated with male infertility. Several recurrent genetic mutations have been reported to be associated with abnormal sperm morphology, but the genetic basis of tapered-head sperm is not well understood. In this study, whole-exome sequencing (WES) identified a homozygous WD repeat domain 12 (WDR12; p.Ser162Ala/c.484T>G) variant in an infertile patient with tapered-head spermatozoa from a consanguineous Chinese family. Bioinformatic analysis predicted this mutation to be a pathogenic variant. To verify the effect of this variant, we analyzed WDR12 protein expression in spermatozoa of the patient and a control individual, as well as in the 293T cell line, by Western blot analysis, and found that WDR12 expression was significantly downregulated. To understand the role of normal WDR12, we evaluated its mRNA and protein expression in mice at different ages. We observed that WDR12 expression was increased in pachytene spermatocytes, with intense staining visible in round spermatid nuclei. Based on these results, the data suggest that the rare biallelic pathogenic missense variant (p.Ser162Ala/c.484T>G) in the WDR12 gene is associated with tapered-head spermatozoa. In addition, after intracytoplasmic sperm injection (ICSI), a successful pregnancy was achieved. This finding indicates that infertility associated with this WDR12 homozygous mutation can be overcome by ICSI. The present results may provide novel insights into understanding the molecular mechanisms of male infertility.
Humans ; Pregnancy ; Female ; Male ; Animals ; Mice ; Teratozoospermia/pathology* ; Semen/metabolism* ; Infertility, Male/metabolism* ; Spermatozoa/metabolism* ; Mutation ; RNA-Binding Proteins/metabolism* ; Cell Cycle Proteins/genetics*