Multiple morphological abnormalities of the flagella (MMAF) represent a severe form of sperm defects leading to asthenozoospermia and male infertility. In this study, we identified a novel homozygous splicing mutation (c.871-4 ACA>A) in the adenylate kinase 7 (AK7) gene by whole-exome sequencing in infertile individuals. Spermatozoa from affected individuals exhibited typical MMAF characteristics, including coiled, bent, short, absent, and irregular flagella. Transmission electron microscopy analysis showed disorganized axonemal structure and abnormal mitochondrial sheets in sperm flagella. Immunofluorescence staining confirmed the absence of AK7 protein from the patients' spermatozoa, validating the pathogenic nature of the mutation. This study provides direct evidence linking the AK7 gene to MMAF-associated asthenozoospermia in humans, expanding the mutational spectrum of AK7 and enhancing our understanding of the genetic basis of male infertility.
Humans ; Male ; Sperm Tail/ultrastructure* ; Homozygote ; Consanguinity ; Asthenozoospermia/pathology* ; Infertility, Male/genetics* ; Mutation ; Pakistan ; Adenylate Kinase/genetics* ; Adult ; Pedigree ; RNA Splicing ; Exome Sequencing ; Spermatozoa

Multiple morphological abnormalities of sperm flagella (MMAF) is a severe form of asthenoteratozoospermia, characterized by morphological abnormalities and reduced motility of sperm, causing male infertility. Although approximately 60% of MMAF cases can be explained genetically, the etiology of the remaining cases is unclear. Here, we identified two novel compound heterozygous variants in the gene, dynein axonemal heavy chain 10 ( DNAH10 ), in three patients from two unrelated Pakistani families using whole-exome sequencing (WES), including one compound heterozygous mutation ( DNAH10 : c.9409C>A [p.P3137T]; c.12946G>C [p.D4316H]) in family 1 and another compound heterozygous mutation ( DNAH10 : c.8849G>A [p.G2950D]; c.11509C>T [p.R3687W]) in family 2. All the identified variants are absent or rare in public genome databases and are predicted to have deleterious effects according to multiple bioinformatic tools. Sanger sequencing revealed that these variants follow an autosomal recessive mode of inheritance. Hematoxylin and eosin (H&E) staining revealed MMAF, including sperm head abnormalities, in the patients. In addition, immunofluorescence staining revealed loss of DNAH10 protein signals along sperm flagella. These findings broaden the spectrum of DNAH10 variants and expand understanding of the genetic basis of male infertility associated with the MMAF phenotype.
Adult ; Humans ; Male ; Alleles ; Asthenozoospermia/pathology* ; Axonemal Dyneins/genetics* ; Dyneins/genetics* ; Exome Sequencing ; Infertility, Male/pathology* ; Mutation ; Pakistan ; Pedigree ; Sperm Tail/pathology*

Asthenoteratozoospermia is one of the most severe types of qualitative sperm defects. Most cases are due to mutations in genes encoding the components of sperm flagella, which have an ultrastructure similar to that of motile cilia. Coiled-coil domain containing 103 (CCDC103) is an outer dynein arm assembly factor, and pathogenic variants of CCDC103 cause primary ciliary dyskinesia (PCD). However, whether CCDC103 pathogenic variants cause severe asthenoteratozoospermia has yet to be determined. Whole-exome sequencing (WES) was performed for two individuals with nonsyndromic asthenoteratozoospermia in a consanguineous family. A homozygous CCDC103 variant segregating recessively with an infertility phenotype was identified (ENST00000035776.2, c.461A>C, p.His154Pro). CCDC103 p.His154Pro was previously reported as a high prevalence mutation causing PCD, though the reproductive phenotype of these PCD individuals is unknown. Transmission electron microscopy (TEM) of affected individuals' spermatozoa showed that the mid-piece was severely damaged with disorganized dynein arms, similar to the abnormal ultrastructure of respiratory ciliary of PCD individuals with the same mutation. Thus, our findings expand the phenotype spectrum of CCDC103 p.His154Pro as a novel pathogenic gene for nonsyndromic asthenospermia.
Asthenozoospermia/pathology* ; Dyneins/genetics* ; Homozygote ; Humans ; Male ; Microtubule-Associated Proteins ; Mutation ; Mutation, Missense ; Sperm Tail/metabolism*

OBJECTIVETo observe the ultrastructural changes of sperm flagella in patients with severe idiopathic asthenospermia.

METHODSUsing the transmission electron microscope, we examined the ultrastructure of sperm flagella from 22 patients with severe idiopathic asthenospermia.

RESULTSUltrastructural anomalies were found in all the 22 patients, 6 with partial or complete absence of internal and external dynamic arms in dedicative of primary ciliary dyskinesia, 1 with hyperplasia, hypertrophy and disordered organization of the fibrous sheath usually referred to as dysplasia of the fibrous sheath, and the other 15 with non-specific flagellar anomalies.

CONCLUSIONExamination of the ultrastructure of sperm flagella in severe idiopathic asthenospermia patients can help to distinguish congenital from acquired flagellar structural anomalies and give valuable guidance in the treatment.

Adult ; Asthenozoospermia ; pathology ; Humans ; Male ; Sperm Tail ; ultrastructure

OBJECTIVE: To evaluate the fertility of infertile males of large samples by the routine semen examination, and to provide a scientific basis for clinical treatment of male infertility and improve efficacy. METHODS: We collected 16 835 semen specimens of infertile man in our hospital (the infertility group), and 1 567 semen specimens of sperm donating candidates from the sperm bank of our hospital (the control group). Semen routine examination was performed by computer-assisted semen analysis (CASA) technology, and the data of semen routine examination were analyzed statistically. RESULTS: The infertile patients with semen abnormalities in our hospital mainly suffered asthenospermia. The proportion of azoospermia, asthenospermia and oligoasthenospermia in the infertility group was higher than that in the control group, while the proportion of oligozoospermia was lower than that in the control group (P<0.001). CONCLUSION Azoospermia, asthenospermia and oligoasthenospermia are related to infertility, but simply oligozoospermia is not necessarily related to infertility.
Asthenozoospermia ; Azoospermia ; China ; Humans ; Infertility, Male ; epidemiology ; Male ; Oligospermia ; Semen Analysis ; Sperm Count ; Spermatozoa ; pathology

OBJECTIVETo study the effects of Wuzi Yanzong Pills (WYP) on sperm mitochondrial membrane potential (MMP) and its ultrastructure in oligo-asthenozoospermia model rats.

METHODSOligo-asthenozoospermia models were made in 50 male rats weighing 200 - 220 g by intragastric administration of Tripterygium Glucosides at 30 mg per kg per d for 8 weeks, and then equally allocated to a model control, a Huangjing Zanyu Capsule (HZC) control, a low-dose WYP, a medium-dose WYP, and a high-dose WYP group. Another 10 age-matched normal male rats were included as normal controls. The rats in the model and normal control groups were given intragastrically distilled water at 10 ml/kg, those in the HZC group administered HZC at 3.01 g/kg, and those in the low-, medium- and high-dose WYP groups medicated with WYP at 2.30, 4.60 and 9.20 g/kg, respectively, once daily for 30 days. At 30 minutes after the last administration, we detected the sperm MMP by JC-1 fluorescent staining and flow cytometry, and examined the sperm ultrastructure under the JEM-1230 transmission electron microscope.

RESULTSJC-1 + % and its fluorescence intensity were (33.77 +/- 6.19)% and 1 468 +/- 496 in the model control, (56.34 +/- 10.35)% and 3 277 +/- 895 in the HZC control, (40.80 +/- 10.40)% and 2 016 +/- 767 in the low-dose WYP, (59.40 +/- 6.51)% and 3 897 +/- 643 in the medium-dose WYP, and (60.71 +/- 7.81)% and 3 371 +/- 647 in the high-dose WYP group, significantly reduced in comparison with (70.80 +/- 4.92)% and 4 360 +/- 945 in the normal control group (P < 0.05), but remarkably higher in the medium- and high-dose WYP groups than in the model controls (P < 0. 05). After modeling, the sperm membrane was loose and degenerated, the mitochondria swelling, variously sized and with incomplete membrane, and the axonemal structure unclear or ruptured. After 30 days of WYP administration, compared with the model control group, the rats exhibited integrated sperm membrane and mitochondrial membrane, reduced mitochondrial swelling and basically normal axonemal and microtubular structures.

CONCLUSIONTripterygium Glucosides could decrease the sperm mitochondrial membrane potential and damage the mitochondrial structure, while WYP could significantly increase the sperm mitochondrial membrane potential and reduce the sperm mitochondrial structure damage. The protection of the integrity of sperm mitochondrial structure and function is one of the mechanisms of WYP acting on oligo-asthenozoospermia.

Animals ; Asthenozoospermia ; pathology ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; Male ; Membrane Potential, Mitochondrial ; drug effects ; Oligospermia ; pathology ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Spermatozoa ; drug effects ; physiology ; ultrastructure

OBJECTIVETo investigate the effects of Yijing Recipe on sperm apoptosis and mitochondrial membrane potential (MMP) in patients with idiopathic oligoathenoteratospermia.

METHODSUsing the self-control method, we examined sperm apoptosis and MMP in 30 patients with oligoathenoteratospermia before and after treated with Yijing Recipe.

RESULTSThe rates of early sperm apoptosis (AV +/PI -) and MMP loss were significantly reduced after treatment as compared with pre-medication ([2.86 +/- 1.47]% vs [4.26 +/- 2.79]% and [21.77 +/- 13.46]% vs [41.73 +/- 20.30]%, P<0.05). No statistically significant difference was observed in the sperm death rate (PI+) before and after treatment ([34.10 +/- 16.26]% vs [30.21 +/- 13.50]%, P>0.05).

CONCLUSIONYijing Recipe can reduce early sperm apoptosis and improve MMP, which may be one of the mechanisms underlying its efficacy on oligoathenoteratospermia.

Adult ; Apoptosis ; drug effects ; Asthenozoospermia ; drug therapy ; pathology ; physiopathology ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Infertility, Male ; drug therapy ; pathology ; physiopathology ; Male ; Membrane Potential, Mitochondrial ; drug effects ; Oligospermia ; drug therapy ; pathology ; physiopathology ; Phytotherapy ; Semen Analysis ; Sperm Motility ; Spermatozoa

OBJECTIVE: To assess how high ligation of varicocele improves sperm DNA integrity in patients with asthenospermia. METHODS: The DNA fragmentation index (DFI) and semen analysis were conducted in 30 subjects with normal sperm samples, and the parameters were used as references. Fifty-six patients from the Department of Urology of the Second Affiliated Hospital of Chongqing Medical University from April 2009 to April 2011 all signed the informed consent. Semen was collected 1 week before the high ligation of varicocele and 3 months after the surgery. We compared the change of the integrity of sperm DNA and semen. RESULTS: Three months after the high ligation of varicocele, DFI and semen indexes significantly improved (P<0.05), but no significant difference was found between open surgery and laparoscopic surgery, unilateral varicocele and bilateral varicocele and Grade I, Grade II, Grade III varicocele (P>0.05). Six months later, 27 spouses were pregnant out of the 53 surgical patients (50.9%). DFI of the pregnancy group and the non-pregnancy group was (13.90±9.70)% and (20.10±10.27)%, with significant different (P<0.05). CONCLUSION Varicocelectomy can improve the sperm DNA integrity in patients with asthenospermia, whose effect is not related to surgical approach, unilateral or bilateral varicocele or varicocele grade.
Adult ; Asthenozoospermia ; etiology ; surgery ; DNA Damage ; DNA Fragmentation ; Humans ; Ligation ; Male ; Middle Aged ; Semen ; Spermatic Cord ; blood supply ; surgery ; Spermatozoa ; pathology ; Urologic Surgical Procedures, Male ; Varicocele ; complications ; surgery ; Young Adult

OBJECTIVETo explore the correlation of the mutation of MTCYB and MTATP6 genes in sperm mitochondria with asthenospermia.

METHODSWe extracted mtDNA from 80 semen samples of asthenospermia and 20 of normal sperm motility, amplified the MTCYB and MTATP6 genes by PCR, and analyzed their mutation by sequencing and BLAST matching.

RESULTSThe deletion of both MTCYB and MTATP6 were detected in 20 of the 80 asthenospermia samples, MTCYB deletion in 16 and MTATP6 deletion in 4, accounting for 20% and 5% respectively. Sequencing and BLAST matching revealed G8887A mutation in the MTATP6 gene in the asthenospermia samples, with a mutation rate of 20%, while no regular mutation was noted in MTCYB. Neither significant deletion nor mutation was observed in any of the 20 samples of normal sperm motility.

CONCLUSIONBoth the deletion and mutation of MTCYB and MTATP6 genes in sperm mitochondria might affect sperm motility in adults.

Adult ; Asthenozoospermia ; genetics ; pathology ; Base Sequence ; Cytochromes b ; genetics ; DNA, Mitochondrial ; genetics ; Humans ; Male ; Mitochondrial Proteins ; genetics ; Mitochondrial Proton-Translocating ATPases ; genetics ; Molecular Sequence Data ; Mutation ; Sequence Homology, Nucleic Acid ; Sperm Count ; Spermatozoa ; metabolism ; pathology

OBJECTIVETo study the relationship between the expression of voltage-dependent Ca2+ channels (VDCCs) cal mRNA and asthenospermia.

METHODSBased on the WHO criteria, we filtered the donated semen by computer-aided sperm analysis (CASA), optimized the ejaculated sperm by discontinuous Percoll grads centrifugation, and examined and quantitated the multitype VDCCs alpha1 mRNA expressions in the sperm of normal men and asthenospermia patients using reverse transcription polymerase chain reaction (RT-PCR).

RESULTSCompared with the normal sperm, there were significant differences among the expressions of alpha1B (39.40 +/- 9.47), alpha1C (48.30 +/- 11.60), alpha1E (3.20 +/- 0.78), alpha1G (8.40 +/- 2.03) and alpha1H (5.70 +/- 1.47) mRNA messages in asthenospermia (P < 0.01).

CONCLUSIONThe abnormal expression of L-type and/or non-L-type VDCCs' mRNAs may be one of the causes of asthenospermia.

Asthenozoospermia ; genetics ; pathology ; Calcium Channels ; genetics ; Gene Expression ; Humans ; Male ; Protein Isoforms ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Semen ; metabolism ; Sperm Motility ; genetics