Testicular cancer seminoma is one of the most common types of cancer among men of reproductive age. Patients with this condition usually present reduced semen quality, even before initiating cancer therapy. However, the underlying mechanisms by which testicular cancer seminoma affects male fertility are largely unknown. The aim of this study was to investigate alterations in the sperm proteome of men with seminoma undergoing sperm banking before starting cancer therapy, in comparison to healthy proven fertile men (control group). A routine semen analysis was conducted before cryopreservation of the samples (n = 15 per group). Men with seminoma showed a decrease in sperm motility (P = 0.019), total motile count (P = 0.001), concentration (P = 0.003), and total sperm count (P = 0.001). Quantitative proteomic analysis identified 393 differentially expressed proteins between the study groups. Ten proteins involved in spermatogenesis, sperm function, binding of sperm to the oocyte, and fertilization were selected for validation by western blot. We confirmed the underexpression of heat shock-related 70 kDa protein 2 (P = 0.041), ubiquinol-cytochrome C reductase core protein 2 (P = 0.026), and testis-specific sodium/potassium-transporting ATPase subunit alpha-4 (P = 0.016), as well as the overexpression of angiotensin I converting enzyme (P = 0.005) in the seminoma group. The altered expression levels of these proteins are associated with spermatogenesis dysfunction, reduced sperm kinematics and motility, failure in capacitation and fertilization. The findings of this study may explain the decrease in the fertilizing ability of men with seminoma before starting cancer therapy.
Acrosin/metabolism* ; Adult ; Case-Control Studies ; Chaperonin Containing TCP-1/metabolism* ; Electron Transport Complex III/metabolism* ; HSP70 Heat-Shock Proteins/metabolism* ; Humans ; Male ; Peptidyl-Dipeptidase A/metabolism* ; Proteasome Endopeptidase Complex/metabolism* ; Proteomics ; Semen Analysis ; Seminoma/metabolism* ; Sodium-Potassium-Exchanging ATPase/metabolism* ; Sperm Count ; Sperm Motility ; Spermatozoa/metabolism* ; Testicular Neoplasms/metabolism*

In this study, we attempted to establish a culture system for in vitro spermatogenesis from spermatogonial stem cells (SSCs) of Bama mini-pig. Dissociated testicular cells from 1-month-old pigs were co-cultured to mimic in vivo spermatogenesis. The testicular cells were seeded in minimum essential medium alpha (α-MEM) supplemented with Knockout serum replacement (KSR). Three-dimensional colonies formed after 10 days of culture. The colonies showed positive staining for SSC-associated markers such as UCHL1, PLZF, THY1, OCT4, Dolichos biflorus agglutinin, and alkaline phosphatase. Induction of SSCs was performed in α-MEM + KSR supplemented with retinoic acid, bone morphogenetic protein 4, activin A, follicle-stimulating hormone, or testosterone. The results showed that STRA8, DMC1, PRM1, and TNP1 were upregulated significantly in the colonies after induction compared to that in testis from 1-month-old pigs, while expression levels of those genes were significantly low compared to those in 2-month-old testis. However, upregulation of ACROSIN was not significant. Replacement of α-MEM and KSR with Iscove's modified Dulbecco's medium and fetal bovine serum did not upregulate expression of these genes significantly. These results indicate that SSCs of Bama mini-pig could undergo differentiation and develop to a post-meiotic stage in α-MEM supplemented with KSR and induction factors.
Acrosin ; Activins ; Alkaline Phosphatase ; Bone Morphogenetic Protein 4 ; Dolichos ; Follicle Stimulating Hormone ; Humans ; In Vitro Techniques ; Infant ; Infant, Newborn ; Spermatogenesis ; Stem Cells ; Swine ; Testis ; Testosterone ; Tretinoin ; Up-Regulation

Objective: To investigate the clinical significance of sperm acrosin activity detection in selecting the method of assisted reproduction for patients with unexplained infertility (UI). METHODS: This retrospective study included 49 UI couples treated by IVFET (49 cycles) after three failures in intrauterine insemination (IUI) and another 95 couples with uterine tube obstruction (UTO) treated by IVF (131 cycles). We analyzed the laboratory data, clinical outcomes and sperm acrosin activity in the two groups of patients. According to the level of sperm acrosin activity of the males, we further divided the UI patients into two subgroups, a < 36 IU/106 sperm group (20 cycles) and a ≥36 IU/106 sperm group (29 cycles), and compared the fertilization rates between the two groups. RESULTS: Compared with UI couples treated by IVFET, the UTO couples treated by IVF had a significantly lower rate of fertilization (67.0% vs 76.4%, P < 0.05) and a higher rate of remedial intracytoplasmic sperm injection (ICSI) (20.4% vs 6.1%, P < 0.05), but showed no statistically significant differences in the rates of MII oocytes, available embryos, highquality embryos, implantation, and clinical pregnancy from the latter group (P >0.05). The sperm acrosin activity was remarkably lower in the UI than in the UTO patients (36.03 vs 61.98 IU/106, P < 0.01), and so was the fertilization rate in the < 36 IU/106 than in the ≥36 IU/106 sperm subgroup (47.7% vs 80.3%, P < 0.01). CONCLUSIONS The low fertilization rate caused by decreased sperm acrosin activity may be the main cause of infertility and the potential factor of UI. When sperm acrosin activity is < 36 IU/106 sperm, IVF plus shortterm fertilization by remedial ICSI should be preferred to IUI.
Acrosin ; analysis ; metabolism ; Embryo Implantation ; Fallopian Tubes ; Female ; Fertilization in Vitro ; methods ; statistics & numerical data ; Humans ; Infertility, Female ; Infertility, Male ; Male ; Pregnancy ; Pregnancy Rate ; Reproduction ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; statistics & numerical data ; Spermatozoa ; metabolism

OBJECTIVETo investigate the progressive motility, (PR), total motility (progressive + non-progressive motility, PR + NP), and acrosin activity of sperm from normal and infertile men at different time points after sperm activation.

METHODSBased on the 5th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen and the results of modified Papanicolaou staining, we divided the semen samples into groups A (normal, n = 28), B (oligoasthenoteratospermia, n = 30), and C (asthenoteratospermia, n = 32). At 1, 24, and 48 hours after sperm activation, we detected sperm PR and PR + NP by CASA and chemical colorimetry, and determined sperm acrosin activity using the modified Kennedy method.

RESULTSSperm PR and PR + NP were significantly decreased in all the three groups at 1-24 hours and even more significantly at 24-48 hours after sperm activation as compared with the baseline (P < 0.05). Sperm acrosin activity showed remarkable reduction in group A (P = 0. 013) , even more significant at 1-24 hours than at 24-48 hours after sperm activation, but not in groups B and C (P = 0.519 and 0.979).

CONCLUSIONSperm PR, PR + NP, and acrosin activity are all decreased with the extension of time after sperm activation, each in a specific manner. Examination of sperm acrosin activity should be applied as a routine tool in the assessment of male fertility.

Acrosin ; metabolism ; Asthenozoospermia ; metabolism ; physiopathology ; Biomarkers ; metabolism ; Humans ; Infertility, Male ; metabolism ; physiopathology ; Male ; Semen ; Sperm Motility ; physiology ; Spermatozoa ; metabolism ; physiology ; Time Factors

OBJECTIVETo investigate the correlation of sperm DNA damage and sperm-nucleoprotein transition with acrosin activity and seminal parameters.

METHODSWe collected 535 semen samples, assessed sperm DNA damage by sperm chromatin dispersion test, and analyzed the correlation of sperm DNA damage and sperm-nucleoprotein transition with acrosin activity and seminal parameters according to the WHO criteria.

RESULTSStatistically significant differences were observed in sperm DNA damage among sperm-nucleoprotein transition, acrosin activity, sperm concentration and the percentage of grade a + b sperm (P < 0.01). Sperm DNA damage was positively correlated with age, sperm-nucleoprotein transition, sperm concentration and the percentage of grade d sperm (P < 0.01 or P < 0.05), but negatively correlated with acrosin activity (P < 0.001). Stepwise linear regression analysis demonstrated that age, sperm concentration, the percentage of grade d sperm, sperm-nucleoprotein transition and acrosin activity were independent variables related to the DNA fragmentation index (DFI). The abnormality rates of sperm-nucleoprotein transition, acrosin activity, sperm concentration and graded a + b sperm were significantly higher in the sperm DNA damage group (DFI > or = 30%) than in the normal control (DFI < 30%) (P < 0.01).

CONCLUSIONSperm DNA damage is closely related with sperm-nucleoprotein transition, acrosin activity and seminal parameters, which may become another important independent parameter for the evaluation of sperm quality.

Acrosin ; genetics ; Adult ; Chromatin ; DNA Damage ; DNA Fragmentation ; Humans ; Infertility, Male ; genetics ; Male ; Nucleoproteins ; genetics ; metabolism ; Sperm Count ; Sperm Motility ; Spermatozoa

OBJECTIVETo investigate the effect of sperm acrosin activity on the IVF-ET outcome.

METHODSWe analyzed sperm parameters, morphology and acrosin activity for 909 infertile husbands by computer-assisted self-assessment (CASA), modified Papanicolaou staining and N-alpha-benzoyl-DL-arginine-p-nitroanilide (BAPNA), respectively, and detected the rates of fertilization, cleavage, quality embryos, embryo cryopreservation, implantation, clinical pregnancy and abortion. The wives were identified as normal or with mere oviduct problems.

RESULTSThe rate of normal sperm morphology and sperm motility, vitality, rapid progressive velocity and concentration were significantly lower in the abnormal acrosin activity group than in the normal one (P < 0.01). Significant positive correlations were observed between acrosin activity and the above-mentioned semen parameters (P < 0.01). There were no significant differences in the number of retrieved eggs, the rates of cleavage, quality embryos, embryo cryopreservation, non-embryo transfer cycles and miscarriages, and the number of transferred embryos between the two groups (P > 0.05). The fertilization rate, the percentage of transfer cycles with only 1 embryo and the rate of implantation and clinical pregnancy were notably higher in the normal acrosin activity group than in the abnormal one (P < 0.01).

CONCLUSIONSperm acrosin activity is closely related with semen parameters, and it helps to predict the sperm fertilizing capacity and IVF-ET outcome.

Acrosin ; metabolism ; Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Infertility, Male ; Male ; Pregnancy ; Pregnancy Rate ; Semen Analysis ; Spermatozoa ; enzymology

OBJECTIVETo evaluate the inhibitory effect of Nandeshi, an acrosin inhibitor, on human acrosin activity.

METHODSWe collected sperm samples from 10 healthy fertile men and cultured them with Nandeshi at 30 degrees C for 5 minutes at the concentrations of 0. 100, 0.120, 0.144, 0.173, 0.207, 0.249, 0.299, 0.358 and 0.430 mmol/L, with the controls treated with a well-known acrosin inhibitor N-alpha-p-tosyl-L-lysine chloromethylketone (TLCK) at 150.0, 189.8, 213.6, 240.3, 270.3, 304.1 and 342.1 mmol/L. Then we determined the residual activity of human acrosin by improved Kennedy assay.

RESULTSThe residual activity of acrosin was negatively correlated with the Nandeshi concentration, and Nandeshi exhibited an inhibition rate about 800 times that of TLCK.

CONCLUSIONNandeshi has a powerful inhibitory effect on human acrosin, and improved Kennedy assay is a simple, practical and highly sensitive technique for the detection of human acrosin activity.

Acrosin ; antagonists & inhibitors ; metabolism ; Contraceptive Agents, Female ; pharmacology ; Enzyme Inhibitors ; pharmacology ; Humans ; Male ; Spermatozoa ; drug effects ; Tosyllysine Chloromethyl Ketone ; pharmacology

AIMTo develop a method for assessing sperm function by measuring released acrosin activity during the acrosome reaction (AR).

METHODSHuman semen samples were obtained from 24 healthy donors with proven fertility after 3-7 days of sexual abstinence. After collection, samples were liquefied for 30 min at room temperature. Standard semen parameters were evaluated according to World Health Organization (WHO) criteria. Calcium ionophore A23187 and progesterone (P4) were used to stimulate the sperm to undergo AR. After treatment, sperm were incubated with the supravital dye Hoechst33258, fixed in a glutaraldehyde-phosphate-buffered saline solution, and the acrosomal status was determined by fluorescence microscopy with fluorescein isothiocyanate-labeled Pisum sativum agglutinin (FITC-PSA). The percentage of sperm undergoing AR (AR%) was compared to sperm acrosin activities as assessed by spectrocolorimetry. The correlation between AR% and acrosin activity was determined by statistical analysis.

RESULTSThe AR% and released acrosin activity were both markedly increased with A23187 and P4 stimulation. Sperm motility and viability were significantly higher after stimulation with P4 versus stimulation with A23187 (P < 0.001). There was a significant positive correlation between released acrosin activity and AR% determined by FITC-PSA staining (r=0.916, P < 0.001).

CONCLUSIONSpectrocolorimetric measurement of released acrosin activity might serve as a reasonable alternative method to evaluate AR.

Acrosin ; physiology ; Acrosome Reaction ; Adult ; China ; Humans ; Male ; Progesterone ; pharmacology ; Semen ; drug effects ; physiology ; Sperm Motility ; drug effects ; physiology

OBJECTIVETo investigate the etiologic factors of globozoospermia.

METHODSRoutine semen analysis, sperm DNA special staining and chromosomal karyotype detection of peripherical blood lymphocytes were performed in a globozoospermia patient.

RESULTSRound-headed spermatozoa were lack of acrosome and the acrosin activity was low. Meanwhile, there was an additional band located in the Y chromosomal short arm.

CONCLUSIONLack of acrosome, low acrosin activity and abnormality of chromosome may be the main reasons for globozoospermia.

Acrosin ; metabolism ; Adult ; Chromosomes, Human, Y ; genetics ; Humans ; Infertility, Male ; etiology ; genetics ; therapy ; Male ; Sex Chromosome Aberrations ; Sperm Injections, Intracytoplasmic ; Spermatozoa ; abnormalities ; ultrastructure

OBJECTIVETo explore the mechanism of interleukin-6 (IL-6) on acrosome reaction (AR) in human sperm.

METHODSSperm acrosin activity was measured by BAEE/ADH and AR evaluated by FITC-PSA.

RESULTSIL-6 could induce the activity of acrosin and superoxide dismutase (SOD) and enhance AR in human sperm. AR was not induced by extracellular Ca2+, and IL-6-induced AR did not occur in Ca2(+)-free medium. Calphostin C, one inhibitor of the protein kinase C (PKC), could block IL-6-induced AR in human sperm.

CONCLUSIONIL-6 could induce AR by stimulating the activity of acrosin and SOD in human sperm, which also involves the activation of PKC, and requires the presence of extracellular Ca2+.

Acrosin ; metabolism ; Acrosome Reaction ; drug effects ; Adult ; Calcium ; pharmacology ; Cells, Cultured ; Humans ; Interleukin-6 ; pharmacology ; Male ; Naphthalenes ; pharmacology ; Protein Kinase C ; antagonists & inhibitors ; Spermatozoa ; drug effects ; enzymology ; Superoxide Dismutase ; metabolism