Mitochondrial Membrane Potential Of Spermatozoa Research Articles

Aging of stallion spermatozoa stored in vitro is delayed at 22°C using a 67mm glucose-10mm pyruvate-based media.

Most commerce of equine seminal doses is carried out using commercial extenders under refrigeration at 5°C. To determine if 10mm pyruvate in a 67mm glucose extender and storage at 22°C could be the basis of an alternative storage method to cooling to 5°C. Stallion ejaculates were extendedin: INRA96 (67mm glucose, non-pyruvate control), modified Tyrode's (67mm glucose-10mm pyruvate), supplemented with 0, 10, 50, and 100μM itaconate. As itaconate was vehiculated in DMSO, a control vehicle was also included. Sperm motility, viability, mitochondrial membrane potential, and production of reactive oxygen species were measured after collection and again after 48 and 96h of storage at 22°C. To disclose molecular metabolic changes, spermatozoa were incubated up to 3h in modified Tyrode's 67mm glucose-10mm pyruvate and modified Tyrode's 67mm glucose, and metabolic analysis conducted. After 96h of storage aliquots stored in the control, INRA96 had a very poor total motility of 5.6%±2.3%, while in the 67mm glucose-10mm pyruvate/10μm itaconate extender, total motility was 34.7%±3.8% (p=0.0066). After 96h, viability was better in most pyruvate-based media, and the mitochondrial membrane potential in spermatozoa extended in INRA96 was relatively lower (p<0.0001). Metabolomics revealed that in the spermatozoa incubated in the high pyruvate media, there was an increase in the relative amounts of NAD+, pyruvate, lactate, and ATP. Aliquots stored in a 67mm glucose-10mm pyruvate-based medium supplemented with 10μM itaconate, maintained a 35% total motility after 96h of storage at 22°C, which is considered the minimum acceptable motility for commercialization. Improvements may be related to the conversion of pyruvate to lactate and regeneration of NAD+.

Rapid thawing of frozen bull spermatozoa by transient exposure to 70 °C improves the viability, motility and mitochondrial health.

Up to now, the definitive conclusion of the positive effects of rapid transient thawing at higher temperatures for shorter durations has not been obtained yet and is still under discussion due to some contradictory findings and limited assessment of post-thawed parameters. The purpose of the current study was to evaluate the effectiveness of rapid thawing in water at 70 °C by using various post-thawed parameters of frozen bull spermatozoa. Experiment 1, monitoring the change of temperature inside frozen bull straw thawed in water at different temperatures. Experiment 2, evaluation of various post-thawed characteristics of frozen bull spermatozoa thawed in water at different temperatures by using a computer-assisted sperm analysis, flow cytometry and immunocytochemistry. The time it took for the temperature inside the straw to warm up to 15 °C was nearly twice as faster when the straw was thawed in 70 °C water compared with 39 °C. Although there were differences among bulls, viability, motility, and mitochondrial membrane potential of spermatozoa thawed at 70 °C for 8 seconds and stabilized at 39 °C for 52 seconds were significantly higher than those of controls (thawed at 39 °C for 60 seconds) at 0 and 3 h after thawing. Just after thawing, however, there were no differences in acrosome integrity and distribution of phospholipase C zeta1, whereas mitochondrial reactive oxygen species production was significantly lower in spermatozoa thawed at 70 °C. From these results, we conclude that rapid thawing at 70 °C and then stabilization at 39 °C significantly improves viability, motility and mitochondrial health of bull spermatozoa rather than conventional thawing at 39 °C. The beneficial effect of rapid transient thawing could be due to shorter exposure to temperatures outside the physiological range, consequently maintaining mitochondrial health.

Use of Biological and Synthetic Polymers for Human Spermatozoa Cryopreservation

BACKGROUND: Spermatozoa cryopreservation is an integral part of the assisted reproductive technologies for treatment of infertility. It is also used to preserve the reproductive potential of men. However, using a standard freezing method with glycerol leads to a decrease in morphological and functional characteristics of spermatozoa in the case of oligoasthenoteratozoospermia (OAT). Therefore, it is relevant to develop effective methods of cryopreservation for such sperm. The use of various biopolymers can stabilize the membrane and bind excess water, which forms ice crystals in the medium that causes cell damage when temperature decreases. OBJECTIVE: To study the effectiveness of using cryoprotectant mixtures based on biological and synthetic polymers [serum albumin, polyvinylpyrrolidone (PVP) and insulin] for the cryopreservation of human spermatozoa with OAT. MATERIALS AND METHODS: Human spermatozoa with OAT were cryopreserved using different cryoprotectant media containing 10 % glycerol or 10 % PVP, 20 % albumin and 1 μg/m L human insulin. The viability, motility and mitochondrial membrane potential of spermatozoa were assessed after rewarming. RESULTS: A cryoprotectant solution containing 10 % PVP, 20 % human serum albumin and 1 μg/m L insulin enabled a similar level (%) of viable gametes compared with the standard method using glycerol, while the number of motile cells was significantly lower (p &lt; 0.008). The membrane mitochondrial potential did not differ significantly from fresh sperm. CONCLUSION: The data obtained in this study show the effectiveness of a biopolymer mixture containing PVP, serum albumin and insulin for the cryopreservation of human OAT spermatozoa.

Association between Fatty Acid Composition, Cryotolerance and Fertility Competence of Progressively Motile Bovine Spermatozoa.

Simple SummarySpermatozoa move forward in the female reproductive tract in a linear pattern defined as progressive motility (PM). PM is associated with fertilization competence and used as a parameter to evaluate the spermatozoa’s quality. The livestock-breeding industry is based on artificial insemination with cryopreserved spermatozoa, making the relationship between PM and cryosurvival important. This study explores the association between PM and spermatozoa’s quality, cryotolerance and fertilization competence. The number of progressively motile spermatozoa within a standard insemination straw positively correlates with conception rate, making progressively motile spermatozoa’s survival during cryopreservation a potentially relevant parameter for spermatozoa evaluation.An association between progressive motility (PM) and spermatozoa fertility competence has been suggested. However, the mechanism that underlies PM is not clear enough. We examined physiological characteristics and fatty acid composition of fresh spermatozoa with high and low PM. Additional analysis of fatty acid composition and structural characteristics was performed on spermatozoa samples with high and low progressively motile spermatozoa’s survival (PMSS), i.e., the ratio between the proportion of progressively motile spermatozoa after and before cryopreservation. Finally, a fertility field trial was conducted to examine the association between the number of PM spermatozoa within the insemination straw post thawing and conception rate. Analysis of fresh spermatozoa revealed a higher omega-6 to omega-3 ratio in ejaculates with low PM relative to those with high PM (p < 0.01). The proportion of polyunsaturated fatty acids was higher in low-PMSS fresh samples (p < 0.05) relative to their high-PMSS counterparts. Fresh samples with high-PMSS expressed a higher mitochondrial membrane potential (p < 0.05) and a higher proportion of viable cells that expressed reactive oxygen species (ROS; p < 0.05). Post-thawing evaluation revealed a reduced proportion of progressively motile sperm, with a prominent effect in samples with high PM relative to low PM, defined before freezing (p < 0.01). No differences in spermatozoa mitochondrial membrane potential or ROS level were found post-thawing. A fertility study revealed a positive correlation between the number of progressively motile spermatozoa within a standard insemination straw and conception rate (p < 0.05). Considering these, the bull PMSS is suggested to be taken into account at the time of straw preparation.

Zinc oxide nanoparticles alter the membrane potential of mitochondria from post-thawed ram spermatozoa

• Zinc oxide nanoparticles increases mitochondrial membrane potential of spermatozoa from ram semen. • Sperm kinetics are not altered by zinc oxide nanoparticles. • Zinc oxide nanoparticles do not protect the plasma membrane of ram sperm during freezing. The objective of this study was to evaluate how the addition of zinc oxide nanoparticles (nano-ZnO) to freezing extender affect the characteristics of post-thawed ram semen. For this research, seminal pools from three Santa Inês rams were used. Each seminal pool was diluted in Tris-egg yolk (5 % glycerol) extender, supplemented with nano-ZnO (0, 10, 50, 100 or 200 μg/mL), at the final concentration of 200 × 10 6 spermatozoa/mL. Subsequently, samples were filled in 0.25-mL straws, frozen in an automated system and stored in liquid nitrogen (−196 °C). At the time of analyses, semen samples were thawed in a water bath at 37 °C for 30 s and processed for evaluation. All samples were evaluated for sperm kinetics by computer-assisted sperm analysis (CASA), as well as assessed for mitochondrial membrane potential (MMP), plasma membrane integrity (PMi) and acrosomal membrane integrity (ACi), by epifluorescence microcopy. The sperm kinetics results showed that when adding nano-ZnO (10, 50, 100 and 200 μg/mL) to freezing extender, there were no differences (P > 0.05) in post-thawed ram semen between control and treatment groups, nor among the treated groups themselves. It is emphasised that all groups treated with nano-ZnO presented more gametes (P ≤ 0.05) with high MMP than the control group. However, no differences were observed (P > 0.05) in the percentage of sperm with PMi between control and treatment groups (nano-ZnO). In conclusion, the addition of nano-ZnO (10, 50, 100 or 200 μg/mL) to freezing extender increases MMP but does not alter the kinetics or protect membranes of ram spermatozoa after the freezing-thawing process.

Ameliorative effect of melatonin on apoptosis, DNA fragmentation, membrane integrity and lipid peroxidation of spermatozoa in the idiopathic asthenoteratospermic men: In vitro.

Fertility loss, recurrent spontaneous abortion and poor outcome in assisted reproductive techniques (ART) have been associated with DNA fragmentation. This work was achieved to evaluate the protective role of melatonin versus apoptosis, DNA fragmentation, membrane integrity and lipid peroxidation of spermatozoa from men with asthenoteratozoospermia (ATS). Some researchers maintain that melatonin can serve as a remedy for apoptosis induction, and it has an impressive effect on boosting the quality and quantity of spermatozoa. For this purpose, semen samples were collected from 50 ATS men and they were divided into control and melatonin (6mM) groups at 2, 4, 6 and 24hr. Concentrating on the reasons for apoptosis is an arduous process, but in the present study for this evaluation mitochondrial membrane potential (MMP), DNA fragmentation by TUNEL and sperm chromatin dispersion (SCD) methods and lipid peroxidation were carried out. Also, sperm viability was performed. In the control group, MDA, TUNEL-positive and SCD were significantly increased but viability and MMP of spermatozoa were significantly decreased. Moreover, in the melatonin group, TUNEL-positive, SCD and MDA levels were significantly decreased and viability and MMP significantly increased, compared to the control group. In outcome, melatonin prescription paves the way for apoptosis down-regulating in the ATS men.

Effect of the oral intake of astaxanthin on semen parameters in patients with oligo-astheno-teratozoospermia: a randomized double-blind placebo-controlled trial.

BackgroundHigher concentrations of seminal reactive oxygen species may be related to male infertility. Astaxanthin with high antioxidant activity can have an impact on the prevention and treatment of various health conditions, including cancer. However, efficacy studies on astaxanthin in patients with oligospermia with/without astheno- or teratozoospermia (O±A±T) have not yet been reported. Our aim was to evaluate the effect of the oral intake of astaxanthin on semen parameters.Patients and methodsIn a randomized double-blind trial, 80 men with O±A±T were allocated to intervention with 16 mg astaxanthin orally daily or placebo. At baseline and after three months basic semen parameters, sperm deoxyribonucleic acid (DNA) fragmentation and mitochondrial membrane potential (MMP) of spermatozoa and serum follicle-stimulating hormone (FSH) value were measured.ResultsAnalysis of the results of 72 patients completing the study (37 in the study group, 35 in the placebo group) did not show any statistically significant change, in the astaxanthin group no improvements in the total number of spermatozoa, concentration of spermatozoa, total motility of spermatozoa, morphology of spermatozoa, DNA fragmentation and mitochondrial membrane potential of spermatozoa or serum FSH were determined. In the placebo group, statistically significant changes in the total number and concentration of spermatozoa were determined.ConclusionsThe oral intake of astaxanthin did not affect any semen parameters in patients with O±A±T.

Cryopreservation Induced Sperm Cryoinjuries in Hariana Bull Semen

Cryopreservation causes breach in the plasma membrane which culminates into loss of membrane permeability, change in membrane fluidity, exudation of acrosomal enzymes, and modification in mitochondrial membrane potential. Keeping these views in mind the current study was conducted to evaluate the apoptosis like changes or cryoinjuries in spermatozoa during cryopreservation. In the present study we collected semen from 4 Hariana bulls and after dilution semen was evaluated for various parameters, thereafter semen was cryopreserved. Various sperm characteristics were further evaluated 24 hrs after cryopreservation. The findings of the current study revealed that there is significant reduction in progressive motility, viability, acrosomal integrity, plasma membrane integrity and mitochondrial membrane potential of spermatozoa after semen cryopreservation. Thus we concluded that cryopreservation induced cryoinjuries in spermatozoa and investigation regarding mechanism involved in these changes is key factor to improve the success of semen cryopreservation.

Biological properties of mouse spermatozoa separated by using the microfluidic sperm sorter

The microfluidic sperm sorter (MFSS) is a novel device that is able to separate spermatozoa with high motility and low DNA fragmentation from raw semen without washing and centrifugation process. However, some biological properties and effect on embryonic development of spermatozoa separated by the MFSS are as yet unexplained. The purpose of this study was to compare and investigate the biological properties and the developmental ability between spermatozoa separated by the MFSS and the swim-up (SU) procedure. Motility parameters and mitochondrial membrane potential (MMP) of spermatozoa prepared by MFSS and SU were studied. Cleavage and blastocyst rate were compared between embryos with spermatozoa separated by MFSS and those with spermatozoa processed by SU. Spermatozoa derived from C57BL/6J mouse were divided into two aliquots. One aliquot was processed by the MFSS and separated into high motility (HM) and low motility (LM) spermatozoa; the second aliquot was centrifuged (1800rpm, 10min) and processed by the swim-up procedure for 20-30 min (SU). Sperm motility parameters of three groups were analyzed by using the sperm motility analyzing system (SMAS). MMP was evaluated with the flow cytometer after JC-1 staining. In addition, spermatozoa of each group were injected into ovulated MII oocytes. After 3.5 hours from ICSI, a part of fertilized oocytes derived from each group was stained immunohistochemically with gamma H2AX antibody to detect DNA damage, and the others were cultured until to the blastocyst stage. By the SMAS analysis, the motile sperm rate was 90.5%, 47.8%, and 89.5% for HM, LM, and SU, respectively. The average speed of HM was significantly higher than that of LM and SU (HM: 116.8±8.6 μm/sec, LM: 68.2±8.2, SU: 83.1±4.1, p<0.05). MMP of spermatozoa in HM and SU were significantly higher than that of LM. After ICSI, there was no significant difference in the rate of pronuclear formation between each group. By immunohistochemical staining, high positive rate of gamma H2AX was observed in male pronucleus of embryos with LM spermatozoa. In addition, the blastocyst rate of embryos with HM was significantly higher than that of LM and was comparable to that of SU (HM: 84.3%, LM: 56.1%, SU: 79.7%, p<0.05). In this study, we demonstrated that spermatozoa with high sperm motility and developmental ability were able to separate by using the MFSS. By optimizing the MFSS system, the most appropriate spermatozoa for IVF/ICSI would be provided efficiently.

Effect of Lepidium meyenii (maca) on testicular function of mice with chemically and physically induced subfertility.

The aim of this study was to evaluate the effect of Lepidium meyenii (maca) in chemically and physically subfertile mice. After 35days, the following groups of mice were evaluated: control, sham, chemical subfertility, chemical subfertility-maca-supplemented, physical subfertility, physical subfertility-maca-supplemented and maca-supplemented only. Motility (32.36%±5.34%) and sperm count (44.4±5.37×10(6) /ml) in the chemically and physically subfertile mice (11.81%±4.06%, 17.34±13.07×10(6) /ml) decreased compared to the control (75.53%±2.97% and 57.4±19.6 10(6) /ml) and sham (53.5%±7.86% and 58.4±14.10 10(6) /ml). Maca was able to reverse the deleterious effect of motility (76.36±1.97) as well as sperm count (53.5±9.18×10(6) /ml) on chemical subfertility. In contrast, maca did not reverse the effects of induced physical subfertility nor motility (18.78%±14.41%) or sperm count (20.17±11.20×10(6) /ml). The percentage of sperm DNA fragmentation in the physically subfertile mice increased (11.1%±19.29%) compared to the control (0.84%±0.85%). However, in the physically subfertile group, maca decreased sperm DNA fragmentation (2.29%±2.30%) closer to the sham (1.04%±0.62%) and the control (0.84%±0.85%). The group supplemented only with maca showed 0.54%±0.50% of spermatozoa with DNA fragmentation. Yet, the differences observed were statistically not significant. In conclusion, it appears that maca activates the cytochrome P450 system after chemically induced subfertility. However, it does not reverse the low mitochondrial membrane potential in spermatozoa compromised in the physical subfertility group.

Effects of the Tris, Tes, or skim milk based extender on in vitro parameters of ram spermatozoa during liquid storage

Cryopreservation can seriously damage structure and physiological function of mammalian spermatozoa. However, liquid storage at reduced temperature may be an alternative of cryopreservation if artificial insemination is performed in relatively short time. In this study, effects of the Tris-hydroxymethyl aminomethane (Tris), N-Tris (hydroxymethyl) -methylaminoethane-sulfonic acid (Tes), or skim milk based extender on ram spermatozoa during liquid storage at 4°C were assessed and compared. Semen samples from 6 adult Yunnan semi-fine wool rams with proven fertility were pooled, equally divided into 3 groups according to extenders, and preserved at 4°C for various durations (0, 1, 3, 5, and 7 days). The motility, moving velocity, and hypo-osmotic swelling capability of spermatozoa were evaluated using a computer assisted sperm analyzer system. The acrosome status, membrane integrity, distribution of phosphatidylserine (PS), and mitochondrial membrane potential (MMP) were analyzed with flow cytometry. The results indicated that following liquid storage in the Tris or Tes based extender for 3 days, the motility, hypo-osmotic swelling capability, and MMP of spermatozoa did not show a significant decrease. Additionally, liquid storage in the Tris or Tes based extender for 24h did not deleteriously affect the moving velocity, acrosome status, and membrane integrity of spermatozoa. However, after liquid storage for 24h, the percentage of spermatozoa with membrane exposed PS was significantly increased. On the contrary, the negative effects of skim milk on the above parameters of ram spermatozoa became more serious compared to the Tris or Tes based extender after liquid storage for 24h (P<0.05). There was no significant difference existing between the Tris and Tes based extender as concerning the parameters evaluated in this study. In conclusion, the Tris or Tes based extender may be more suitable for liquid storage of ram spermatozoa at reduced temperature compared to the skim milk based extender. Tris can be substituted by Tes for liquid storage of ram semen. The future study should focus on assessment of molecular changes and fertility of ram spermatozoa after liquid storage.

Simultaneous evaluation of plasma membrane integrity, acrosomal integrity, and mitochondrial membrane potential in bovine spermatozoa by flow cytometry.

The present study aimed to develop an objective evaluation procedure to estimate the plasma membrane integrity, acrosomal integrity, and mitochondrial membrane potential of bull spermatozoa simultaneously by flow cytometry. Firstly, we used frozen-thawed semen mixed with 0, 25, 50, 75 or 100% dead spermatozoa. Semen was stained using three staining solutions: SYBR-14, propidium iodide (PI), and phycoerythrin-conjugated peanut agglutinin (PE-PNA), for the evaluation of plasma membrane integrity and acrosomal integrity. Then, characteristics evaluated by flow cytometry and by fluorescence microscopy were compared. Characteristics of spermatozoa (viability and acrosomal integrity) evaluated by flow cytometry and by fluorescence microscopy were found to be similar. Secondly, we attempted to evaluate the plasma membrane integrity, acrosomal integrity, and also mitochondrial membrane potential of spermatozoa by flow cytometry using conventional staining with three dyes (SYBR-14, PI, and PE-PNA) combined with MitoTracker Deep Red (MTDR) staining (quadruple staining). The spermatozoon characteristics evaluated by flow cytometry using quadruple staining were then compared with those of staining using SYBR-14, PI, and PE-PNA and staining using SYBR-14 and MTDR. There were no significant differences in all characteristics (viability, acrosomal integrity, and mitochondrial membrane potential) evaluated by quadruple staining and the other procedures. In conclusion, quadruple staining using SYBR-14, PI, PE-PNA, and MTDR for flow cytometry can be used to evaluate the plasma membrane integrity, acrosomal integrity, and mitochondrial membrane potential of bovine spermatozoa simultaneously.

Seminal plasma aids the survival and cervical transit of epididymal ram spermatozoa

Seminal plasma purportedly plays a critical role in reproduction, but epididymal spermatozoa are capable of fertilisation following deposition in the uterus, calling into question the biological requirement of this substance. Through a combination of direct observation of spermatozoa in utero using probe-based Confocal Laser Endomicroscopy, in vivo assessment of sperm fertility and in vitro analysis of various sperm functional parameters, this study investigated the role of seminal plasma in spermatozoa transit through the cervix of the ewe. Following deposition in the cervical os, epididymal spermatozoa previously exposed to seminal plasma displayed an enhanced ability to traverse the cervix as evidenced by both significantly higher pregnancy rates and numbers of spermatozoa observed at the utero-tubal junction when compared with epididymal spermatozoa not previously exposed to seminal plasma. The beneficial effect of seminal plasma on sperm transport was clearly localised to transit through the cervix as pregnancy rates of spermatozoa deposited directly into the uterus were unaffected by exposure to seminal plasma. This phenomenon was not explained by changes to sperm motion characteristics, as seminal plasma had no effect on the motility, kinematic parameters or mitochondrial membrane potential of spermatozoa. Rather, in vitro testing revealed that seminal plasma improved the ability of epididymal spermatozoa to penetrate cervical mucus recovered from ewes in oestrus. These results demonstrate that the survival and transport of ram spermatozoa through the cervix of the ewe is not linked to their motility or velocity but rather the presence of some cervical penetration trait conferred by exposure to seminal plasma.

Seminal plasma aids the survival and cervical transit of epididymal ram spermatozoa.

Seminal plasma purportedly plays a critical role in reproduction, but epididymal spermatozoa are capable of fertilisation following deposition in the uterus, calling into question the biological requirement of this substance. Through a combination of direct observation of spermatozoa in utero using probe-based Confocal Laser Endomicroscopy, in vivo assessment of sperm fertility and in vitro analysis of various sperm functional parameters, this study investigated the role of seminal plasma in spermatozoa transit through the cervix of the ewe. Following deposition in the cervical os, epididymal spermatozoa previously exposed to seminal plasma displayed an enhanced ability to traverse the cervix as evidenced by both significantly higher pregnancy rates and numbers of spermatozoa observed at the utero-tubal junction when compared with epididymal spermatozoa not previously exposed to seminal plasma. The beneficial effect of seminal plasma on sperm transport was clearly localised to transit through the cervix as pregnancy rates of spermatozoa deposited directly into the uterus were unaffected by exposure to seminal plasma. This phenomenon was not explained by changes to sperm motion characteristics, as seminal plasma had no effect on the motility, kinematic parameters or mitochondrial membrane potential of spermatozoa. Rather, in vitro testing revealed that seminal plasma improved the ability of epididymal spermatozoa to penetrate cervical mucus recovered from ewes in oestrus. These results demonstrate that the survival and transport of ram spermatozoa through the cervix of the ewe is not linked to their motility or velocity but rather the presence of some cervical penetration trait conferred by exposure to seminal plasma.

Alterations in mitochondria membrane potential and oxidative stress in infertile men: a prospective observational study

Objective To evaluate the mitochondrial membrane potential (MMP) of spermatozoa and its correlation with semen parameters and production of reactive oxygen species (ROS) in infertile men and healthy donors. Design Controlled prospective study. Setting Male infertility clinic, Glickman Urological Institute, The Cleveland Clinic Foundation, Cleveland, Ohio. Patient(s) Nineteen infertile men and 7 healthy volunteers. Intervention(s) Standard semen analysis, assessment of MMP and ROS production in spermatozoa. The MMP was assessed by flow cytometry using the probe carbocyanine DiOC 6(3) and ROS was measured with chemiluminescence assay using luminol. Main outcome measure(s) The results of MMP are reported as the median interquartile range (IQR) number of cells counted in different areas of fluorescence. Results of ROS measurement are expressed as ×10 6 counted photons per minute per 20 million sperm (cpm). Results The patients with abnormal semen parameters had a significantly lower MMP [1337.7 (1066.38, 1879.2)], and higher ROS [1.12 (0.26, 3.86)] than the donors [MMP: 2482.9 (2162.5, 3520.6)] and [ROS: 0.10 (0.01, 0.14)]. The MMP was positively correlated with sperm concentration (r = 0.62) and negatively correlated with the ROS produced (r = −0.45). Conclusion(s) Measuring MMP in spermatozoa provides useful information about a man's fertility potential. Increased ROS production by spermatozoa is associated with a decreased MMP.