OBJECTIVETo investigate the expression of carbonic anhydrase II (CA2) in human testes and spermatozoa, and to compare the expressions of CA2 in ejaculated spermatozoa between normozoospermic and asthenozoospermic men.

METHODSThe localization of CA2 in human testes was observed by immunohistochemistry, and that in human sperm by immunofluorescence. Western blot was used to detect the expression of CA2 in the semen samples obtained from 16 normozoospermic and 16 asthenozoospermic volunteers.

RESULTSThe CA2 protein was shown to be localized in the tail of elongating spermatids by immunohistochemistry and in the flagellum of human sperm by immunofluorescence. Western blot revealed an obviously increased expression of CA2 in the spermatozoa of asthenozoospermic patients, with statistically significant difference from the normozoospermic group (1.84 +/- 0.32 vs 1.41 +/- 0.26, P < 0.05).

CONCLUSIONThe CA2 protein is expressed in the spermatogenic stage of elongating spermatids in human testes and localized in the sperm tail. The expression of CA2 is significantly increased in the spermatozoa of asthenozoospermic men, which might be responsible for low sperm motility.

Asthenozoospermia ; metabolism ; Carbonic Anhydrase II ; metabolism ; Humans ; Male ; Sperm Motility ; Spermatozoa ; metabolism ; Testis ; metabolism

OBJECTIVETo investigate the role of the cation channel of sperm 1 (CatSper1) protein in the pathogenesis of idiopathic asthenozoospermia.

METHODSSperm samples from patients with idiopathic asthenozoospermia were separated by Percoll discontinuous density gradients, and the distribution and expression of the CatSper1 protein were determined by immunocytochemistry. Western blotting was used to detect the different expressions of CatSper1 in the ejaculated sperm from the normal control, mild asthenozoospermia, moderate asthenozoospermia and severe asthenozoospermia groups, followed by statistical analyses.

RESULTSThe expression of CatSper1, located in the principle piece of the sperm tail, was reduced significantly in the samples from the idiopathic asthenozoospermia patients as compared with the normal controls (t = 2.188, P = 0.042). The relative contents of the CatSper1 protein in the sperm of the control, mild asthenozoospermia, moderate asthenozoospermia and severe asthenozoospermia groups were 0.806 +/- 0.266, 0.669 +/- 0.207, 0.505 +/- 0.214 and 0.295 +/- 0.162, respectively, significantly decreased in the asthenozoospermia patients in comparison with the normal controls (P <0.05). There was a positive correlation between the percentage of progressively motile sperm and the relative content of the CatSper1 protein (r = 0.633, P = 0.000).

CONCLUSIONThe decreased or abnormal expression of the CatSper1 protein may be a factor involved in the pathogenesis of idiopathic asthenozoospermia.

Adult ; Asthenozoospermia ; metabolism ; Calcium Channels ; metabolism ; Case-Control Studies ; Humans ; Male ; Spermatozoa ; metabolism ; Young Adult

OBJECTIVETo compare the expressions of ODF1 (outer dense fiber of the sperm tail 1) in ejaculated spermatozoa from normozoospermic and asthenozoospermic men with low sperm motility.

METHODSSemen analyses were performed on the semen samples obtained from normozoospermic (n=20) and asthenozoospermic (n=20) volunteers according to the WHO criteria. To rule out the contamination of germ cells and leucocytes, the human ejaculated spermatozoa were purified by a discontinuous Percoll density gradient centrifugation. RT-PCR and Western blot were used to detect the expressions of ODF1 in the spermatozoa from the two groups.

RESULTSRT-PCR showed that the expression of ODF1 mRNA was significantly lower in the spermatozoa from the asthenozoospermic patients than in those from the normozoospermic men (1.35 +/- 0.25 vs. 2.79 +/- 0.28, P < 0.05). Western blot confirmed the results from RT-PCR and revealed an obviously decreased expression of ODF1 in the spermatozoa of the asthenozoospermic patients, with statistically significant difference from the normozoospermic group (1.44 +/- 0.26 vs. 3.64 +/- 0.34, P < 0.05).

CONCLUSIONThe expression of ODF1 was significantly decreased in the ejaculated spermatozoa of asthenozoospermic men, which might be responsible for low sperm motility.

Asthenozoospermia ; metabolism ; Heat-Shock Proteins ; metabolism ; Humans ; Male ; Sperm Motility ; Spermatozoa ; metabolism

OBJECTIVETo investigate the role of the SEPT4 protein in the pathogenesis of idiopathic asthenozoospermia.

METHODSSamples of ejaculated sperm from idiopathic asthenozoospermia patients and normozoospermic men were separated and purified by Percoll discontinuous density gradients, the distribution and expression of SEPT4 in the sperm samples were determined by immunocytochemistry, and the expressions of SEPT4 mRNA and SEPT4 protein were detected by RT-PCR and Western blot.

RESULTSImmunocytochemistry showed that the expression of SEPT4, located in the annulus, was significantly reduced in the sperm of the idiopathic asthenozoospermia patients (t = 3.452, P < 0.01). RT-PCR revealed that the expression of SEPT4 mRNA was significantly lower in the sperm of the idiopathic asthenozoospermia patients than in those of the normozoospermic men (t = 3.521, P < 0.05). Western blot confirmed the results of RT-PCR (t = 5.872, P < 0.05).

CONCLUSIONThe expression of SEPT4 is significantly decreased in the ejaculated sperm of idiopathic asthenozoospermia patients, which might be one of the causes of idiopathic asthenozoospermia.

Adult ; Asthenozoospermia ; metabolism ; Case-Control Studies ; Humans ; Male ; Septins ; metabolism ; Sperm Motility ; Spermatozoa ; metabolism ; Young Adult

ObjectiveTo investigate the expression and location of IQ motif-containing G (IQCG) in the human testis, compare its expression in normal-motility sperm with that in the sperm of asthenospermia patients, and explore its possible mechanisms and its correlation with fertility.

METHODSThe expression of the IQCG gene in the human testis was detected by RT-PCR and its location in the testis and sperm was determined by immunohistochemistry and immunofluorescence staining. Semen samples were collected from normal males, patients with asthenospermia, and fertile men that succeeded in artificial insemination with donor's sperm (AID), followed by analysis of the IQCG protein expression in different groups of samples by Western blot.

RESULTSImmunohistochemistry showed that IQCG was extensively expressed in the human testis, in the spermatocytes and spermatids, specifically in the sperm tail, weakly expressed or absent in the spermatogonial stem cells, and strongly expressed in the spermatogonial cells. The expression of IQCG was significantly lower in the asthenospermia patients than in the normal males (P= 0.041). Western blot manifested that IQCG was expressed in the semen of all the three groups of subjects, with statistically significant differences between the normal men and severe asthenospermia patients (P = 0.032) as well as between the fertile males and the severe asthenospermia group (P = 0.027) .

CONCLUSIONSIQCG may act on human sperm motility and its abnormal expression possibly reduces sperm motility and fertility. An insight into its action mechanisms may shed some new light on the etiology and treatment of asthenospermia.

Asthenozoospermia ; etiology ; metabolism ; therapy ; Fertility ; Humans ; Male ; Semen ; Sperm Motility ; Spermatozoa ; Testis ; metabolism

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 investigate the mRNA and protein expression levels of cysteine-rich secretory protein 2 (CRISP2) in the sperm of asthenospermia patients, and explore their relationship with sperm motility and related molecular mechanism.

METHODSWe collected 78 semen samples from adult male patients with asthenospermia and another 70 from healthy volunteers as controls. We extracted total RNA and total protein from the sperm following purification of the sperm by Percoll gradient centrifugation, and detected the relative expressions of CRISP2 mRNA and protein in the two groups by RT-PCR, SYBR Green real-time PCR and Western blot.

RESULTSThe expression of CRISP2 mRNA was down-regulated by 4.3 times and that of the CRISP2 protein by 1.71 times in the asthenospermia patients, significantly lower than in the normal control group (P < 0.05).

CONCLUSIONThe down-regulation of CRISP2 mRNA and protein expressions in the sperm of asthenospermia patients may be closely related with decreased sperm motility, which suggests that CRISP2 may serve as a potential molecular target for the research of asthenospermia.

Adult ; Asthenozoospermia ; genetics ; metabolism ; Case-Control Studies ; Glycoproteins ; genetics ; metabolism ; Humans ; Male ; Sperm Motility ; Spermatozoa ; metabolism ; physiology

OBJECTIVETo investigate the role of the TEKT4 protein in the pathogenesis of idiopathic asthenozoospermia.

METHODSWe separated and purified the ejaculated sperm from idiopathic asthenozoospermia patients and normozoospermic men by Percoll discontinuous density gradients, and detected the distribution and the expressions of TEKT4 mRNA and TEKT4 protein by RT-PCR and Western blot.

RESULTSRT-PCR revealed that the expression of TEKT4 mRNA was significantly lower in the sperm of the idiopathic asthenozoospermia patients than in those of the normozoospermic men (0.59 +/- 0.13 vs 0.75 +/- 0.15, t = 4.325, P < 0.05), and Western blot confirmed the results of RT-PCR (0.48 +/- 0.14 vs 0.69 +/- 0.13, t = 5.939, P < 0.05).

CONCLUSIONThe expression of TEKT4 is significantly decreased in the ejaculated sperm of idiopathic asthenozoospermia patients, which might be one of the causes of idiopathic asthenozoospermia.

Adult ; Asthenozoospermia ; metabolism ; Blotting, Western ; Case-Control Studies ; Cytoskeletal Proteins ; metabolism ; Humans ; Male ; RNA, Messenger ; genetics ; Sperm Motility ; Spermatozoa ; metabolism

OBJEVTIVETo investigate the expression of cysteine-rich secretory protein 2 (CRISP2) in spermatozoa of patients with asthenozoospermia and explore its clinical significance.

METHOSSemen samples were collected from 24 normal volunteers and 24 patients with asthenozoospermia for detecting CRISP2 mRNA and protein expressions using qRT-PCR and Western blotting, respectively. The correlation of CRISP2 expressions with sperm morphology, progressive motility and fertility prognosis were analyzed in patients with asthenozoospermia.

RESULTSCRISP2 protein expression was obviously lowered in the ejaculated spermatozoa of patients with asthenozoospermia as compared to the normal volunteers, but no significant difference in CRISP2 mRNA expression was found between the two groups. Correlation analysis showed that CRISP2 protein expression was positively correlated with normal sperm morphology (r=0.6182, P=0.0037) and progressive motility (r=0.6309, P=0.0029). Follow-up study of the patients revealed a higher fertility rate in patients with a relatively high CRISP2 protein expression than in those with low CRISP2 protein expression (80.0% vs 20.0%, P=0.0230).

CONCLUSIONThe expression level of CRISP2 protein is positively correlated with normal sperm morphology and progressive motility. A reduced CRISP2 protein expression indicates poor fertility prognosis of patients with asthenozoospermia, suggesting the potential value of CRISP2 as a novel therapeutic target for treating asthenozoospermia.

Asthenozoospermia ; metabolism ; Case-Control Studies ; Fertility ; Follow-Up Studies ; Glycoproteins ; metabolism ; Humans ; Male ; RNA, Messenger ; Sperm Motility ; Spermatozoa ; metabolism

OBJECTIVETo study the differentially expressed genes in asthenospermia to gain a deeper insight into the molecular mechanisms of the disease.

METHODSWe analyzed the differentially expressed genes in asthenospermia using GATHER, PANTHER and ToppGene online bioinformatics tools.

RESULTSOur bioinformatics mining and analyses revealed that the differentially expressed genes in asthenospermia played important roles in the cellular protein and macromolecular metabolism, protein modification, cell death, cell apoptosis and apoptosis induction.

CONCLUSIONAsthenospermia patients experience a decline in sperm activity and the basic life activities of sperm simultaneously, and are also prone to cell apoptosis or death. Such differentially expressed genes as KIF3B, MYO15A, KIF6, KIF26B, KIF3A, DNHD2, DMN, DYNC2H1, STARD9, MYOHD1, and TPM1, which are involved in cytoskeletal structure, microtubule movement and cell movement, may be associated with asthenospermia, and therefore deserve further studies.

Asthenozoospermia ; genetics ; metabolism ; Computational Biology ; Databases, Genetic ; Gene Expression Profiling ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; Spermatozoa ; metabolism