Human Sperm Motility Research Articles

In vitro ameliorative effects of ellagic acid on vitality, motility and DNA quality in human spermatozoa.

Oxidative stress (OS) plays a significant role in the etiology of male infertility, resulting in the impairment of male reproduction. This condition, characterized by an imbalance in the levels of oxidizing and antioxidant species in the seminal fluid, has a harmful impact on sperm functions and DNA integrity. The present study aimed to evaluate the anti-genotoxic action of ellagic acid, a polyphenolic molecule of natural origin having a powerful antigenotoxic, anti-inflammatory and antiproliferative role. An OS condition was induced in vitro by incubating normozoospermic human semen samples in benzene for 45, 60 and 90 min. DNA integrity was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling assay, RAPD-PCR was performed to calculate the genome template stability, while the percentage of intracellular reactive oxygen species (ROS) was assessed by the 2', 7'-dichlorofluorescein assay. Our results showed that ellagic acid has a consistent protective effect on DNA integrity, as well as on sperm vitality and motility, by counteracting generation of intracellular ROS. The results of this study suggest ellagic acid as a suitable molecule to protect sperm DNA from oxidative stress, with a potentially significant translational impact on the management of the male infertility.

The sodium/proton exchanger SLC9C1 (sNHE) is essential for human sperm motility and fertility

Asthenozoospermia, defined by the absence or reduction of sperm motility, constitutes the most frequent cause of human male infertility. This pathological condition is caused by morphological and/or functional defects of the sperm flagellum, which preclude proper sperm progression. While in the last decade many causal genes were identified for asthenozoospermia associated with severe sperm flagellar defects, the causes of purely functional asthenozoospermia are still poorly defined. We describe here the case of an infertile man, displaying asthenozoospermia without major morphological flagellar anomalies and carrying a homozygous splicing mutation in SLC9C1 (sNHE), which we identified by whole-exome sequencing. SLC9C1 encodes a sperm-specific sodium/proton exchanger, which in mouse regulates pH homeostasis and interacts with the soluble adenylyl cyclase (sAC), a key regulator of the signalling pathways involved in sperm motility and capacitation. We demonstrate by means of RT-PCR, immunodetection and immunofluorescence assays on patient's semen samples that the homozygous splicing mutation (c.2748 + 2 T > C) leads to in-frame exon skipping resulting in a deletion in the cyclic nucleotide-binding domain of the protein. Our work shows that in human, similar to mouse, SLC9C1 is required for sperm motility. Overall, we establish a homozygous truncating mutation in SLC9C1 as a novel cause of human asthenozoospermia and infertility.

8 Effects of magnetic-activated cell sorting on human sperm motility and DNA fragmentation index

Selection of spermatozoa before their use for assisted reproductive techniques is an important step in therapy of human infertility. The DNA fragmentation index of sperm plays a major role in pregnancy rates following IVF and intracytoplasmic sperm injection (ICSI). Sperm analyses and standard sperm selection methods in many cases do not eliminate a sufficient proportion of sperm with apoptosis and DNA fragmentation. Magnetic-activated cell sorting (MACS) is a selection method that eliminates apoptotic spermatozoa based on the presence of externalized phosphatidylserine residues. The aim of our study was to evaluate the effect of MACS on human sperm motility and DNA fragmentation index (DFI) in a patient population. The participants were 63 male patients of an IVF clinic, 34 to 45 years old, with 3 years of primary infertility due to male factor. Semen analysis was performed according to the World Health Organization guidelines (2010) and revealed oligoasthenoteratozoospermia in 63 patients. The DFI of fresh semen samples was evaluated using the sperm chromatin structure assay (SCSA) test and revealed DFI 32.4±5.9%. The SCSA test was done on a flow cytometer CyFlow Space (Sysmex-Partec; Evenson 2016 Anim. Reprod. Sci. 169, 56-75; https://doi.org/10.1016/j.anireprosci.2016.01.017). Sperm motility was studied on Hamilton Thorne IVOS. For MACS, we used the MACS® ART Annexin V system (Miltenyi Biotec). The semen sample was diluted to a concentration 10×106 spermatozoa mL−1. After double-density gradient centrifugation, the pellet was resuspended in 100µL of MACS Art Annexin V reagent and added MACS Art Binding Buffer (BB) to 500µL. The sample was gently mixed and incubated for 15min at room temperature. After rinsing the column with BB, the sperm-bead suspension was added on top with BB and, immediately after that, the annexin V-negative and annexin V-positive fractions were obtained (MiniMACS; Miltenyi Biotec). Data were evaluated by ANOVA Student’s t-test. Fresh semen samples collected from the patients had an average sperm concentration of 29.7±5.7×106 mL−1, motility of 32.7±5.9%, and DFI of 32.4±5.9%. Motility of spermatozoa after MACS for the annexin-negative fraction was 47.2±6.3% and for the annexin-positive fraction was 3.5±2.3% (P<0.003). Similarly, the annexin-negative spermatozoa had a lower DFI (10.5±3.8%) rate than did the annexin-positive fraction (67.8±5.9%; P<0.003). The MACS technique allowed a significant reduction of DNA fragmentation levels (from 32.4% for the original sample to 10.5% for the annexin-negative; P<0.01). The separation of a distinct population of nonapoptotic spermatozoa with intact membranes may optimize outcomes from IVF and ICSI procedures. Magnetic-activated cell sorting of human sperm using annexin-V microbeads results in selection of a population with enhanced motility and reduced DFI rates.

Addition of peroxiredoxin 6 (PRDX6) to IVF fertilization medium maintains motility and longevity of human spermatozoa

This study aims to investigate the protective effects of peroxiredoxin 6 on the total motility and progressive motility of human spermatozoa. Semen samples with normal parameters were collected from 23 males and supplemente... | Find, read and cite all the research you need on Tech Science Press

Erythropoitein Increases In Vitro Motility and Vitality of Human Spermatozoa.

Erythropoietin and its receptor are expressed in the male reproductive system. Initial studies have shown that erythropoietin affects the motility of spermatozoa. The aim of the present study was to investigate the in vitro effect of erythropoietin in the motility and vitality of human spermatozoa. Forty-three semen samples, obtained after 2-4 days of abstinence from sex, were analyzed and processed using density gradient centrifugation. Aliquots containing one million of spermatozoa were treated with either erythropoietin, at concentrations of 10 and 100 mIU/μl or standard culture medium for one hour. Progressive motility and vitality of spermatozoa significantly increased following treatment with erythropoietin. The effect was not dose-dependent. The supplementation of culture medium with erythropoietin improves sperm processing in terms of vitality and motility. Future research should focus on the effects of erythropoietin on sperm capacitation as well as on the signal transduction pathways activated by erythropoietin and its receptor in spermatozoa.

Expression of PPP3CC and not PPP3R2 is associated with asthenozoospermia

Background and objective: Protein phosphatase 3 catalytic subunit gamma (PPP3CC) and protein phosphatase 3 regulatory subunit B, beta (PPP3R2) are respectively the catalytic and regulatory subunits of calcineurin in sperm. Deficiency in either protein causes impaired sperm motility leading to male infertility. Many cases of sterility are attributed to asthenozoospermia (AZS); however, it remains unknown whether PPP3CC and PPP3R2 are related to AZS. Material and methods: Quantitative PCR and Western blotting were used to investigate the expression levels of PPP3CC and PPP3R2 in the spermatozoa of patients with AZS and to explore the clinical significance. Results: Two calcineurin inhibitors cyclosporine A (CsA) and tacrolimus (FK506) markedly impaired the total motility and progressive motility of human sperm, indicating that PPP3CC or PPP3R2 might be involved in AZS. PPP3CC mRNA and protein expression was lower in the ejaculated spermatozoa of patients with AZS than in normal sperm (NS). Correlation analysis showed that PPP3CC protein expression correlated positively with progressive motility (r = 0.2592, P < 0.05); however, there were no significant differences in PPP3R2 mRNA and protein levels between AZS and NS. Conclusion: These findings suggest that the abnormal expression of PPP3CC rather than PPP3R2 might be a pathological factor or indicator in AZS. Thus, PPP3CC may be a potential therapeutic or diagnostic target for some cases of male infertility.

The synthesis of a novel pentoxifylline derivative with superior human sperm motility enhancement properties

mPTF1 demonstrated lower genotoxicity, higher affinity for PDEs, & reasonable pharmacokinetic, druglikeness & ADMET properties compared to Pentoxifylline.

Testis developmental related gene 1 (TDRG1) encodes a progressive motility-associated protein in human spermatozoa.

Is there an association between the human testis-specific gene, testis developmental related gene 1 (TDRG1) and human sperm motility? TDRG1 is associated with asthenozoospermia and involved in regulating human sperm motility. Many testis-specific proteins potentially regulate spermatogenesis and sperm motility. We have identified a novel human testis-specific gene, TDRG1, which encodes a 100-amino-acid protein localized in the human sperm tail, yet little is known about its role in human spermatozoa. Sperm samples were obtained from normozoospermic men and asthenozoospermic men who visited the reproductive medical center at Jiangxi Maternal and Child Health Hospital, Nanchang, Jiangxi, China between February 2018 and January 2019. In total, 27 normozoospermic men and 25 asthenozoospermic men were recruited to participate in the study. The level of TDRG1 in sperm of normozoospermic and asthenozoospermic men was examined by immunoblotting and immunofluorescence assays. Progressive motility was examined by computer-aided sperm analysis. The correlation between the TDRG1 protein level and progressive motility was analyzed by linear regression. TDRG1 was imported into the sperm of normozoospermic and asthenozoospermic men using a cell-penetrating peptide (CPP)-fused TDRG1 recombinant protein (CPP-TDRG1), and the progressive motility was examined. Also, the altered proteins associated with TDRG1 in asthenozoospermic sperm were detected using label-free quantification method-based quantitative proteomic technology. TDRG1-interacting proteins were identified by co-immunoprecipitation coupled with tandem mass spectrometry analysis. The mean level of TDRG1 was significantly decreased in sperm of asthenozoospermic men compared with normozoospermic men (P < 0.05) and was positively correlated with percentage of progressively motile sperm (r2 = 0.75, P = 0.0001). The introduction of TDRG1 into human sperm, using CPP, significantly increased progressive motility (P < 0.05) and improved the progressive motility of sperm from asthenozoospermic men to the normal level. TDRG1 forms a protein complex with sperm-motility related proteins in human sperm and its downregulation was associated with a decrease in other motility-related proteins. N/A. The sample size was limited and larger cohorts are needed for verifying the positive effect of CPP-TDRG1 on human sperm motility. Furthermore, the caution should be paid that a comprehensive safety examination would be performed to evaluate whether CPP-TDRG1 is a possible treatment approach for asthenozoospermia. Our results provide new insights into the mechanisms of sperm motility which may contribute to the diagnosis and treatment for asthenozoospermia. National Natural Science Foundation of China (81501317 and 81871207 to H.C.; 81771644 to T.L.; 31671204 to X.Z.; 81571432 to Y.T.). The authors have no conflicts of interest to declare.

Quinoline Derivative Enhances Human Sperm Motility and Improves the Functional Competence

In this study, we aimed to explore the beneficial properties of novel quinoline derivatives on human sperm motility and its functional competence. Nine novel quinoline derivatives were screened for their effect on motility in human spermatozoa from normozoospermic ejaculates. Compounds with impressive sperm motility enhancement properties were further assessed for their effect on functional competence of human spermatozoa. To determine the effect on the fertilizing ability of spermatozoa processed with quinoline derivatives and to assess developmental competence of embryos derived, in vitro fertilization (IVF) was performed using mouse model. Among the nine quinoline derivatives, 2 compounds (6MQT and 2,6DQT) exhibited significant enhancement in sperm progressive motility and survival at 24 h. Further, non-significant increase in curvilinear velocity (VCL), straight line velocity (VSL), and amplitude of lateral head displacement (ALH) was observed. Capacitation, intracellular cAMP level and tyrosine phosphorylated sperm proteins were significantly higher in 6MQT (P < 0.05) and 2,6DQT (P < 0.001) compared to control. In vitro fertilization (IVF) experiments using Swiss albino mice revealed that spermatozoa processed with 6MQT had non-significantly higher blastocyst rate and a superior blastocyst quality, while, 2,6DQT resulted in significantly lower blastocyst rate (P < 0.05) compared to control. Quinoline derivative 6MQT has significant motility enhancement property under in vitro conditions.Graphical abstract

Effect of pyruvate on human sperm &lt;i&gt;in vitro&lt;/i&gt;

The study objective is to investigate the biological effect of energy substrate (pyruvate) on human sperm motility in vitro. Materials and methods . The study was conducted using human sperm from 14 ejaculate samples obtained by the standard method. Motility and viability were evaluated per the World Health Organization guidelines under 400x magnification. After dilution, 1 ml was sampled from the ejaculate for the experiment and control. The experiment consisted of addition of pyruvate to the test tube with the ejaculate in final concentration of 1 mmol. Results. The level of initial sperm motility corresponded to normozoospermia. The percentage of progressive motile sperm increased by 24, 67 and 84 % after 30, 60 and 120 min of the experiment. The percentage of non-progressive motile sperm and total percentage of motile sperm showed no significant change during this time period. The number of live spermatozoa showed no significant change during the experiment. Conclusion. The study showed that pyruvate can stimulate motility of human sperm in vitro.

Green tea extract as a cryoprotectant additive to preserve the motility and DNA integrity of human spermatozoa.

Cryopreservation impairs sperm quality and functions, including motility and DNA integrity. Antioxidant additives in sperm freezing media have previously brought improvements in postthawed sperm quality. Green tea extract (GTE) is widely considered as an excellent antioxidant, and its beneficial role has been proven in other human cells. This study aims to evaluate the GTE as a potential additive in cryopreservation media of human spermatozoa. In part one, the semen of 20 normozoospermic men was used to optimize the concentration of GTE that maintains sperm motility and DNA integrity against oxidative stress, induced by hydrogen peroxide (H2O2). Spermatozoa were treated with GTE at different concentrations before incubation with H2O2. In part two, the semen of 45 patients was cryopreserved with or without 1.0 ng ml−1 GTE. After 2 weeks, the semen was thawed, and the effect on sperm motility and DNA fragmentation was observed. Our data showed that GTE significantly protected sperm motility and DNA integrity against oxidative stress induced by H2O2 when added at a final concentration of 1.0 ng ml−1. We found that the addition of 1.0 ng ml−1 GTE to cryopreservation media significantly increased sperm motility and DNA integrity (both P < 0.05). More interestingly, patients with high sperm DNA damage benefited similarly from the GTE supplementation. However, there was no significant change in the reactive oxygen species (ROS) level. In conclusion, supplementing sperm freezing media with GTE has a significant protective effect on human sperm motility and DNA integrity, which may be of clinical interest.

Recent updates of sperm cryopreservation technique: a literature review

Background: Nowadays, the decline in male fertility is a significant concern due to the mechanical, systems disturbances, genetic, diet, lifestyle, and environment. Currently, no strategies for medical protection of the germinal epithelium are available. Sperm cryopreservation technique has been established to overcome those issues. Accordingly, this study aims to informed the recent updates of sperm cryopreservation technique from kinds of literature. Methods: A review of relevant literature was performed to elaborate the recent updates in the sperm cryopreservation technique. A total of 45 qualified published literature of all years until 2019 were collected from several electronic database and manual search and included in this review.Results: According to the literature, the first record of sperm cryopreservation was made by Lazzaro Spallanzani in 1776. He used to snow during winter and observed that the motility of human spermatozoa could be preserved after freezing and thawing. Sperm cryopreservation programs provide temporarily or permanently infertile men an opportunity to seek paternity. Sperm cryopreservation is the most effective and the most strongly recommended option for fertility preservation prior to any treatment or occupational gonadotoxic exposures. Sperm cryopreservation should be discussed and offered prior to therapy for malignant diseases. Freezing sperm before the initiation of treatment provides patients with future fertility potential.Conclusion: Sperm cryopreservation could be offered to the patient before the vasectomy procedure in order to prevent reversal vasectomy or surgical sperm retrieval in the future.Â

The effect of vitamin B12 supplement on post‐thaw motility, viability and DNA damage of human sperm

Sperm cryopreservation may lead to adverse effects on sperm structure and function. Cyanocobalamin (vitamin B12) has antioxidant potential and can protect DNA from free radical-induced damages. Recent studies have shown that vitamin B12 preserves glutathione that leads to modulate oxidative stress responses. Also, vitamin B12 might act directly as a scavenger of ROS. The aim of this study was to investigate the effects of vitamin B12 supplementation on human sperm parameters during the cryopreservation process. Thirty semen samples were obtained from normozoospermic men. Using cryopreservation medium supplemented with different concentrations of vitamin B12 (0, 0.5, 1, 2, 2.5mg/ml), the semen samples were cryopreserved. After thawing, all samples were evaluated for motility and viability. Based on results, 2mg/ml was considered as the optimal concentration of vitamin B12 for evaluating sperm DNA fragmentation. The results showed that 1 and 2mg/ml vitamin B12 significantly increased post-thawing motility and viability compared with the 0mg/ml vitamin B12 (p<.05). Also, by supplementing with 2mg/ml vitamin B12, DNA fragmentation decreased when compared to the control. The present study showed that cryopreservation medium supplemented with vitamin B12 at 2mg/ml couldimprove sperm quality after freeze-thaw process.

Hypotonic challenge reduces human sperm motility through coiling and folding of the tail.

Human ejaculates collected for in vitro procedures show variably rapid increases in osmolality, depending on enzymatic degradation of compounds. Changes in osmolality can affect cell functions due to the energy consuming processes needed to control cell volume. The aim was to examine the effects of a hypotonic challenge for spermatozoa exposed to increased osmolality. Spermatozoa were selected by density gradient centrifugation and washed in media with different osmolalities. Osmolality was measured by freezing‐point depression and sperm velocities by CASA. Swimming pattern observations and assessments of tail morphology of fixed spermatozoa were done with phase contrast microscopy. Increased osmolality did not change the curvilinear velocity (VCL), while decreased osmolality reduced or abolished VCL nonreversibly. For spermatozoa first exposed to 400 mOsm/kg, reversal of osmolality to 290 mOsm/kg reduced the VCL and the average path velocity (VAP) permanently. Hypotonic challenges increased sperm tail coiling and folding in a dose‐response pattern. Spermatozoa once adjusted to high osmolality in the liquefied ejaculate are likely to suffer if exposed to a medium with a lower osmolality. For improved success of Assisted Reproductive Technologies (ART), it appears to be important to minimise the duration of sperm exposure to the ejaculate, by early dilution or sperm preparation.

Mitochondrial Activation and Reactive Oxygen-Species Overproduction during Sperm Capacitation are Independent of Glucose Stimuli.

Spermatozoa capacitation is a complex process that requires specific ionic and energetic conditions to support biochemical alterations leading to motility hyperactivation. However, human sperm capacitation is still poorly understood. Herein, we studied the effects of glucose on human sperm capacitation. Healthy men seminal samples (n = 55) were submitted to a density gradient centrifugation and incubated in capacitating conditions in the absence or presence of increasing glucose concentrations (0, 5.5, 11, and 22 mM). Viability and total motility were accessed. Phosphotyrosine levels were measured. Mitochondrial activity and endogenous ROS production were evaluated. Oxidative stress-induced damage was analyzed. Culture media was collected and analyzed by 1H-NMR. Our results show that glucose is essential for human sperm capacitation and motility. Notably, we observed that mitochondrial activity increased even in the absence of glucose. This increased mitochondrial activity was followed by a ROS overproduction, although no oxidative stress-induced damage was detected. Our results show that glucose is essential for capacitation but mitochondrial activation is independent from its stimuli. ROS overproduction may take part on a finely regulated signaling pathway that modulates or even activates capacitation. Taken together, our results constitute a paradigm shift on human sperm capacitation physiology.

Aspirin decreases human sperm motility and vitality, chelates seminal calcium, but insignificantly reduces seminal nitric oxide production

Lately, there is a systematic research consensus that reveals adverse effects of aspirin on semen quality characteristics; however, such consensus is lacking further confirmation by human studies. Therefore, here, we asked whether sperm motility and vitality are affected in the presence of aspirin at 0.1 and 1mM in the ejaculated semen, and whether such effect may be due to an alteration in seminal calcium ions or seminal nitric oxide production. Forty-three semen samples from different normozoospermic men were recruited in this study. Sperm motility was measured by Makler counting chamber, and sperm vitality was measured by Eosin test. Calcium chelating effect of aspirin and seminal nitric oxide production was measured spectrophotometrically. Aspirin at both tested concentrations significantly (p<.05) reduced progressive grade-a motility and vitality of spermatozoa. Additionally, aspirin was found to have significant ability (p<.05) to bind seminal calcium ions, but insignificantly reduced the amount of seminal nitric oxide. In conclusion, sperm motility and vitality were reduced in the presence of aspirin at 0.1 and 1mM in semen. Such reduction may be attributable to the ability of aspirin to chelate seminal calcium ions, but not to an alteration in the amount of nitric oxide produced.

Pantoprazole, a proton-pump inhibitor, impairs human sperm motility and capacitation in vitro.

The effects of PPIs on human sperm fertilizing capacity were poorly investigated although these drugs are widely over-used. Two publications retrospectively studied relationships between any PPI intake and sperm parameters from patients consulting at infertility clinics, but the conclusions of these reports were contradictory. Only two reports investigated the effects of lansoprazole and omeprazole on sperm motility and found lansoprazole to be deleterious and omeprazole to be neutral for sperm motility. The inconsistency of the PPI effect in the previous reports emphasizes the need for more basic research on human spermatozoa, taking into account the hypothesis that the different PPI drugs may have different effects on sperm physiology. Do PPIs, which are among the most widely sold drug in the word, impact negatively human sperm capacitation and sperm motility? The effects of PPIs on human sperm maturation and motility were analyzed by CASA, flow cytometry, and Western blot. We tested the impact of 6 different PPIs on human sperm motility and capacitation. We showed that pantoprazole, but not the other PPIs, decreased sperm progressive motility and capacitation-induced sperm hyperactivation. We therefore investigated further the effects of pantoprazole on sperm capacitation, and we observed that it had a significant deleterious effect on the capacitation-induced hyperpolarization of the membrane potential and capacitation-associated protein phosphorylation. Our results indicate that exposure to pantoprazole has an adverse effect on the physiological competence of human spermatozoa. As the capacitation process takes place within the female tract, our results suggest that PPIs intake by the female partner may impair in vivo sperm maturation and possibly fertilization. Moreover, the absence of adverse effect by PPIs on mouse sperm emphasizes the need to develop reprotox assays using human material to better assess the effects of medication intake on sperm physiology.

Phthalates and infertility: an issue in hernia meshes?

Background: Based on recent data that sperm motility in humans might be (temporarily) affected when lightweight meshes are used for laparoscopic inguinal hernia repair, the aim of this study was to identify the plasticizers present in meshes used for hernia repair. Methods: Mesh samples (n = 13) were cut into small pieces (1 cm × 1 cm), and, after the addition of internal standard, plasticizers were extracted with hexane:acetone (1:1; v:v) by vortex and ultrasonication. The analysis was performed by gas chromatography and mass spectrometry (GC-MS) in electron ionization mode (EI) with full scan acquisition. The identification of compounds was done by injection of reference standards and using the Wiley mass spectral library. Results: None of the targeted plasticizers, including phthalates, were present in the analyzed mesh samples. In particular, di-(2-ethylhexyl) phthalate (DEHP), a major phthalate plasticizer frequently used in medical devices in the past, was not detected in any of the samples. Conclusion: The implantation of mesh materials affecting male fertility, whether temporarily or not, cannot be explained by the presence of the targeted plasticizers, since none of them were found in the wide range of commonly used meshes that were tested in our study.

Aquaporin3 expression and the potential role of aquaporins in motility and mitochondrial membrane potential in human spermatozoa.

Aquaporins are water selective channels which play important roles in cell volume regulation during the transmission of spermatozoa to female tract. This study investigated the expression of aquaporin3 and determined the role of aquaporins in human sperm motility and mitochondrial membrane potential (MMP). RT-PCR and flow cytometry analysis were done to investigate aquaporin3 expression levels, and immunolocalisation of aquaporin3 in the spermatozoa was detected using immunocytochemical analysis. The sperm suspension was divided into four groups of spermatozoa: (a) Spermatozoa at 0hr, (b) spermatozoa in control group, (c) spermatozoa treated with HgCl2 (as an aquaporin inhibitor) and (d) spermatozoa treated with HgCl2 + and 2-mercaptoethanol. The sperm samples were examined in terms of sperm motility and mitochondrial membrane potential. Results confirmed aquaporin3 expression in human spermatozoa and immunocytochemistry results showed an intense immunoreactivity in whole sperm tail. After 60min, HgCl2 showed a significant decrease in motility and MMP compared to the control group. At this time point, 2-mercaptoethanol in the HgCl2 + 2-mercaptoethanol group reversed the effects of HgCl2 as compared to the HgCl2 group. Present study showed the expression and immunolocalisation of AQP3 in human spermatozoa and the potential role of AQPs in the sperm motility and MMP.

SPANX-A/D protein subfamily plays a key role in nuclear organisation, metabolism and flagellar motility of human spermatozoa

Human sperm protein associated with the nucleus on the X chromosome (SPANX) genes encode a protein family (SPANX-A, -B, -C and -D), whose expression is limited to the testis and spermatozoa in normal tissues and to a wide variety of tumour cells. Present only in hominids, SPANX-A/D is exclusively expressed in post-meiotic spermatids and mature spermatozoa. However, the biological role of the protein family in human spermatozoa is largely unknown. Combining proteomics and molecular approaches, the present work describes the presence of all isoforms of SPANX-A/D in human spermatozoa and novel phosphorylation sites of this protein family. In addition, we identify 307 potential SPANX-A/D interactors related to nuclear envelop, chromatin organisation, metabolism and cilia movement. Specifically, SPANX-A/D interacts with fumarate hydratase and colocalises with both fumarate hydratase and Tektin 1 proteins, involved in meeting energy demands for sperm motility, and with nuclear pore complex nucleoporins. We provide insights into the molecular features of sperm physiology describing for the first time a multifunctional role of SPANX-A/D protein family in nuclear envelope, sperm movement and metabolism, considered key functions for human spermatozoa. SPANX-A/D family members, therefore, might be promising targets for sperm fertility management.