Affect Sperm Motility Research Articles

The effect of astaxanthin on human sperm parameters after cryopreservation.

The aim of the study was to examine the protective efficacy of astaxanthin (ASTA) against the damage that occurs during sperm cryopreservation. This experimental study was carried out on waste semen samples of 30 normozoospermic individuals who applied for semen analysis. Semen samples were divided into four equal volumes and 0 μM (control group), 50 μM, 100 μM, and 500 μM ASTA were added to each group. All groups were stored frozen in a liquid nitrogen tank. Semen samples were removed from liquid nitrogen after 72 hours and were thawed. Motility evaluation of sperm was performed. In addition, sperm was stained with acidic aniline blue to detect DNA chromatin condensation. The highest motility loss was found in the control group and the least motility loss was in the 100 μM ASTA group. When examined in terms of sperm chromatin condensation, condensed sperm count was higher in the 100 μM ASTA group than in the other groups. It has been observed that ASTA added to the cryoprotectant substance during sperm cryopreservation positively affects sperm motility and reduces the number of decondensed sperm.

Effect of quercetin or butylated hydroxytoluene on cooled or frozen-thawed ram sperm quality

Cooling and freezing processes cause physical and chemical damage to sperm by cold shock and oxidative stress. This study aimed to evaluate the effect of two antioxidants on sperm parameters of cooled and frozen-thawed ram semen diluted in an egg yolk-based extender. Semen was collected from 30 rams and processed in two consecutive experiments to test the inclusion of different concentrations of quercetin and butylated hydroxytoluene (BHT) in an egg yolk-based semen extender. Dimethyl sulfoxide (DMSO) was added as a solvent to the semen extender in a ratio of 1 mL DMSO for 90 mg of quercetin and 1 mL DMSO for 880 mg of BHT. After collection, semen was diluted at 200 × 106 motile sperm/mL (control) and split into different groups in each experiment. In experiment 1, semen was diluted with the extender containing quercetin (Q5, 5 μg/mL; Q10, 10 μg/mL; Q15, 15 μg/mL) or DMSO alone (DMSO1, 0.055 μL DMSO per mL; DMSO2, 0.165 μL DMSO per mL). In experiment 2, semen was diluted with the extender with BHT (BHT1, 0.5 μg/mL; BHT2, 1 μg/mL; BHT3, 1.5 μg/mL) or DMSO alone (DMSO3, 0.375 μL DMSO per mL; DMSO4, 1.125 μL DMSO per mL). After dilution, the semen was divided into two aliquots. Treated ram sperm samples were also subjected to different storage methods. The first set of samples was cooled at 5 °C for 24 h, whereas the second set of samples was frozen-thawed. Sperm motility parameters and plasma membrane integrity (PMI) were evaluated immediately after dilution (0h) and 24 h after cooling and in the frozen-thawed samples via computer-assisted sperm analysis and epifluorescence microscopy, respectively. The inclusion of quercetin or BHT did not affect sperm motility parameters or PMI of fresh, cooled, or frozen-thawed sperm in this study (P < 0.05). However, further studies are needed to test the effects of these antioxidants on the fertility of cryopreserved ram semen.

The therapeutic effect of co-administration of pentoxifylline and zinc in men with idiopathic infertility.

Pentoxifylline is a derivative of methylxanthine that affects sperm motility. Also, zinc is an antioxidant that is involved in the activation of antioxidant enzymes. This study aimed to evaluate the effect of co-administration of pentoxifylline, and zinc in men with idiopathic infertility. In the present study, men with idiopathic infertility were identified and randomly divided into four groups: pentoxifylline, zinc, pentoxifylline + zinc, and placebo. According to the grouping, the patients received pentoxifylline and zinc for 3months. Then, sperm parameters, biochemical factors, reproductive hormones, inflammatory factors, and DNA damage were evaluated before and after intervention. Data analysis was performed using SPSS software. Pentoxifylline and zinc were significantly effective in improving biochemical parameters, inflammatory factors, concentration, and motility of sperm. Pentoxifylline did not affect sperm morphology and reproductive hormones. However, in the zinc and zinc + pentoxifylline groups, a significant increase in normal morphology and reproductive hormones was observed. In the pentoxifylline group, sperm DNA fragmentation increased significantly, while in the zinc and zinc + pentoxifylline group, DNA fragmentation reduced significantly. Because of the role of zinc in protecting sperm chromatin, it is recommended that zinc and pentoxifyllinebe prescribed simultaneously. Clinical trial code: NCT05156684.

Association of Exposure to Particulate Matter Air Pollution With Semen Quality Among Men in China

The health effects of exposure to particulate matter (PM) on male fertility remain unclear. Although PM exposure has been linked with semen quality, the results were inconsistent. To examine the association of different size fractions of PM (<2.5 μm [PM2.5], 2.5-10 μm [PM2.5-10], and ≤10 μm [PM10]) exposure with semen quality in China. This retrospective cohort study was conducted using data on men whose wives underwent assisted reproductive technology procedures in a tertiary hospital in Shanghai, China, from January 1, 2013, to December 31, 2019. Participants were from 340 prefecture-level cities of China and were followed up from the date of their first visit until December 31, 2019. Data were analyzed from December 1, 2020, to May 15, 2021. Exposure to PM2.5, PM2.5-10, and PM10 during the entire period (0-90 days before semen ejaculation) and key periods of sperm development (0-9, 10-14, and 70-90 days before semen ejaculation). Semen quality, including sperm count, concentration, and motility. A total of 33 876 men were included in the final analysis, with a mean (SD) age of 34.1 (5.7) years and large variation of PM exposure. For example, the median exposure to PM2.5 during the entire period of sperm development was 46.05 (IQR, 34.38-61.65) μg/m3. During the entire period of sperm development, exposures to higher levels of PM were significantly associated with lower total and progressive sperm motility. For total sperm motility, an IQR increase in PM2.5 exposure was associated with an estimated effect decrease of -3.60% (95% CI, -3.93% to -3.26%); an increase in PM2.5-10 exposure, with an estimated effect decrease of -0.45% (95% CI, -0.76% to -0.14%); and an increase in PM10 exposure, with an estimated effect decrease of -2.44% (95% CI, -2.91% to -1.96%). Similar results were observed for progressive motility. An IQR increase in PM2.5 or PM10 exposures was associated with an estimated effect decrease on sperm progressive motility of -1.87% (95% CI, -2.37% to -1.36%) and -1.05% (95% CI, -1.45% to -0.64%), respectively. However, no significant associations were observed between PM exposure during the entire period of sperm development and sperm count or concentration. These findings suggest that PM exposure may adversely affect sperm motility and highlight the need to reduce ambient particulate air pollution exposure for reproductive-aged men.

Dysregulation of NrF2 expression mediates testicular injury and infertility in 3-monochloro-1,2-propandiol-intoxicated rats with special reference to accessory gland-related pathology.

3-Monochloropropane-1,2-diol (3-MCPD) is a food contaminant formed during acid hydrolysis of vegetable proteins. The toxicological evaluation of smaller doses of 3-MCPD is essential for safety evaluation of this compound. The present study investigates the toxicologic potential of 3-MCPD on male genital organs of rats, applies a correlation between the induced infertility and developed lesions in testes, epididymis, and accessory glands and study the possible mechanisms of 3-MCPD-induced male infertility. Forty rats were randomly divided into four main groups of ten animals each: the control untreated group and three treated groups that were orally administered 3-MCPD at different doses (3, 7.5 and 15mg/kg b.w) daily via stomach intubation for five successive days per week. Five rats from each group were euthanized after 30days. The remaining rats were euthanized after 90days to establish subacute and chronic toxicity studies. Oxidative stress markers, Nrf2 gene expression, semen analysis, and histopathological examination were performed at the end of each experimental period. Results indicated that 3-MCPD induces infertility in male rat via disruption of Nrf2 expression in the testicular tissue with subsequent increased oxidative stress indicators in the testis that affect spermatogenesis and induced testicular degeneration, in addition, induction of epididymal lesions that affect sperm motility and concentration and finally possible development of hyperplastic tissue reactions in accessory glands of intoxicated rats predicting the carcinogenic potential of this compound.

In vitro fertilization using sperm activated by ML-2-3 isolated from Morinda lucida Bentham leaves.

PurposeML‐2–3 is a novel tetracyclic iridoid derived from Morinda lucida Bentham leaves. This compound has anti‐trypanosomal and anti‐leishmanial effects. In this study, the authors investigated effects of ML‐2–3 on in vitro fertilization (IVF) rates, motility, and acrosome reaction of the mouse sperm.MethodsIVF was performed using sperm from BALB/cByJJcl mice treated with ML‐2–3. Computer‐assisted sperm analysis (CASA) was performed on the sperm of C57BL/6J mice to investigate sperm motility. The effect of ML‐2–3 on the acrosome reaction was examined by observing the fluorescence of sperm labeled with the acrosin‐EGFP transgene.ResultsML‐2–3 improved IVF in BALB/cByJJcl mice with low fertilization rates. The optimum concentration of ML‐2–3 in sperm pre‐culture medium was 20 µM, and no significant toxicity of ML‐2–3 was observed in developing embryos at this concentration. ML‐2–3 affected sperm motility but not the acrosome reaction. ML‐2–3 increased the IVF rate of mouse sperm that had been refrigerated for 3 days.ConclusionsML‐2–3 can improve the outcome of IVF and motility without inducing the acrosome reaction in mice. These effects of ML‐2–3 on sperm behaviors are different from those of the similar drugs.

Bilateral Retractile Testis: A Possible Risk Factor for Male Infertility

Healthy youth is an asset and strength for economic and social growth of any society. Developmental changes during puberty of youngsters need proper guidance to avoid future physical, physiological and psychological ailments. Bilateral Retractile Testis (BRT) is a health problem if remains unnoticed till adolescence can lead to tribulations. BRT may affect testicular parenchyma and spermatogenesis. Cremasteric muscles (muscles attached with testes) pull the testes up into groin so it randomly moves up and down and do not reside in its natural pouch always. It has been a controversial issue in male infertility as retraction may affect infertility. Objective: To estimate the prevalence of male infertility among patients with Bilateral Retractile Testis. Methodology: A total of 200 male patients were enrolled after the power calculation, with different reproductive health problems between the age of 18 - 40 years during the year 2016 at Reproductive health clinic Jinnah Post Graduate Medical Center (JPMC), Karachi. 25 male patients were reported with infertility and BRT. The data was collected using self-developed questionnaire including outdoor patients’ physical check-up, manual testicular examination as size and shape were done. Patient height, weight, blood pressure, their lifestyle, and dressing regarding wearing tight jeans habits were asked. Semen analysis was carried out showing abnormality in sperm count, motility or morphology and entered and analysed in the SPSS version 10. Means and percentages were calculated. Result: Out of 200 male infertile patients, a total of 25 (12.5%) patients with a diagnosis of BRT were found in the sample with the mean age of 29 years. Moreover, it was identified that those having BRT also have low motility of the sperms. Based on the results it was concluded that BRT is a possible cause of male infertility in Pakistani population. Conclusion: BRT is a possible risk factor to affect sperm motility and low sperm motility can lead to male infertility. The study estimates the prevalence of 12.5% among our sampled men.

Effects of mobile phone use on semen parameters: a cross-sectional study of 1634 men in China

Mobile phones play an irreplaceable role in modern people's lives. However, the radiofrequency electromagnetic radiation produced by mobile phones has also caused increasing concern. A cross-sectional study was conducted to investigate the effect of radiofrequency electromagnetic radiation produced by mobile phones on semen parameters in 1634 men who underwent semen examination at the Department of Reproductive Endocrinology, Women's Hospital, School of Medicine, Zhejiang University, China. Analysis of variance and multivariate linear regression were used to explore differences among different groups. A P <0.05 was considered statistically significant. The results showed significant associations among different groups of daily mobile phone use time and daily duration of phone calls in the percentage of progressively motile spermatozoa (P =0.004 and P =0.007), rapid progressively motile spermatozoa (P =0.012 and P =0.006) and total motile spermatozoa (P =0.004 and P =0.046). After adjustments for the confounding effects of age and body mass index by multiple linear regression, the results showed that the daily duration of mobile phone use had a negative effect on sperm motility. However, there was no statistically significant correlation between daily phone call duration and sperm motility. Therefore, the daily duration of mobile phone use may negatively affect sperm motility and impair male fertility.

Effects of exogenous hormones treatment on spermiation and plasma levels of gonadal steroids in Roughscale sole, Clidoderma asperrimum

Roughscale sole (Clidoderma asperrimum) is only wild caught because basic reproductive research on this species is lacking and gamete production in an artificial setting has not been successful. Exogenous hormone treatment has been used to induce gonadal maturation and final spermiation in wild-caught individuals. In this study, the effects of an exogenous hormone on spermiation in roughscale sole was investigated by implanting different concentrations of a salmon gonadotrophin-releasing hormone analog (sGnRH; 0, 25, 50, and 100 μg/kg body weight) into male fishes. The control group did not produce sperm after 21 days post-implantation, and the duration of spermiation was shorter compared to the other groups. The spermiation period and milt amount differed among the hormone-treated groups according to the hormone concentration used. Milt volumes in the groups treated with 25 and 100 μg/kg sGnRH increased compared to the control group, whereas exogenous hormone treatment had no effect on the movable sperm ratio. The spermatocrit was high at the beginning of spermiation in all groups and then tended to decrease gradually over time except in the experimental group treated with 100 μg/kg sGnRH. Plasma levels of testosterone, 11-ketotestosterone, 17α, 20β-dihydroxy-4-pregnen-3-one were not significantly affected by the sGnRH treatments. Our results demonstrate that it is possible to prolong the spermiation period and increase milt volume by treating male roughscale soles with an exogenous hormone. In addition, the artificial hormone treatment did not affect sperm motility.

MicroRNA-222 Transferred From Semen Extracellular Vesicles Inhibits Sperm Apoptosis by Targeting BCL2L11.

Seminal plasma contains a large number of extracellular vesicles (EVs). However, the roles of these EVs and their interactions with sperm are not clear. To identify the important molecules affecting sperm motility in EVs, we analyzed RNA from seminal plasma EVs of boars with different sperm motility using whole-transcriptome sequencing and proteomic analysis. In total, 7 miRNAs, 67 lncRNAs, 126 mRNAs and 76 proteins were differentially expressed between the two groups. We observed that EV-miR-222 can obviously improve sperm motility. In addition, the results suggested that miR-222 was transferred into sperm by the EVs and that miR-222 affected sperm apoptosis by inhibiting the expression of EGFR, BCL2L11, BAX, CYCs, CASP9 and CASP3. The results of electron microscopy also showed that overexpression of miR-222 in EVs could reduce sperm apoptosis. The study of the whole transcriptomes and proteomes of EVs in boar semen revealed some miRNAs may play an important role in these EVs interactions with Duroc sperm, and the findings suggest that the release of miR-222 by semen EVs is an important mechanism by which sperm viability is maintained and sperm apoptosis is reduced. Our studies provide a new insight of miR-222 in EVs regulation for sperm motility and sperm apoptosis.

Epididymal mRNA and miRNA transcriptome analyses reveal important genes and miRNAs related to sperm motility in roosters

Sperm motility is a crucial trait in chicken production, and the epididymis is an essential organ in the reproductive system. Currently, the molecular mechanisms underlying sperm motility in the epididymis are unclear. In this study, 8 cDNA libraries and eight miRNA libraries were constructed from roosters (4 chickens per group) with diverse sperm motility. After a comparative analysis of epididymal transcriptomes, we detected 84 differentially expressed genes (DEGs) using the edgeR package. Integrated interpretation of DEGs indicated that MMP9, SLN, WT1, PLIN1, and LRRIQ1 are the most promising candidate genes affecting sperm motility in the epididymis of roosters. MiR-146a, mir-135b, and mir-205 could play important regulatory roles in sperm maturation, capacitation, and motility. Additionally, a comprehensive analysis of the mRNA and miRNAs transcriptomes in silico identified a promising gene-miRNA pair miR-135b-HPS5, which may be a vital regulator of sperm motility in the epididymis. Our findings provide novel integrated information of miRNAs and genes that shed light on the regulatory mechanisms of fertility in roosters.

Protein kinases regulate hyperactivated motility of human sperm.

Protein kinases regulate hyperactivated motility of human sperm.

Vitamin D3 Supplementation Effects on Spermatogram and Oxidative Stress Biomarkers in Asthenozoospermia Infertile Men: a Randomized, Triple-Blind, Placebo-Controlled Clinical Trial.

The objective is to evaluate the effects of vitamin D3 (VD3) on sperm parameters and seminal and serum oxidative stress (OS) biomarkers in asthenozoospermia infertile men. This randomized, triple-masking, placebo-controlled clinical trial conducted on 86 asthenozoospermia infertile men with serum 25 hydroxy vitamin D3 (25-OH-D3) < 30ng/ml in the infertility clinic of Ahvaz Jahad Daneshgahi, Iran. Patients were randomly allocated to groups A and B, who received daily 4000IU vitamin D3 (VD3) and matching placebo respectively for 3months. Demographic data, dietary intake, physical activity, sun exposure, anthropometric indices, serum and seminal levels of MDA (Malondialdehyde), 8-hydroxy-2- Dioxy Guanosine (8-OHDG), total antioxidant capacity (TAC) and calcium, sperm DNA fragmentation index (DFI), serum 25-OH-D3, parathyroid hormone (PTH), phosphorus, and sperm parameters were assessed. VD3 supplementation had no significant effects on body weight, body mass index (BMI), waist circumference (WC), body fat (BF), 8-OHDG, DFI, semen volume, sperm count, and normal sperm morphology, but increased post-intervention mean and mean change of serum 25-OH-D3 (P < 0.001, P < 0.001), PTH (P < 0.001, P < 0.001) and phosphorus (P = 0.009, P = 0.049) and seminal calcium (P = 0.035, P = 0.038) and serum calcium (P = 0.008, P = 0.009), seminal TAC (P < 0.001, P < 0.001), and serum TAC (P = 0.007, P = 005), total sperm motility (P < 0.001, P < 0.001) and progressive sperm motility (P < 0.001, P < 0.001) and decreased seminal MDA (P = 0.017, P = 0.004) and serum MDA (P = 006, P = 0.005) significantly compared to the baseline and placebo group respectively. VD3 supplementation may modulate OS and affect sperm motility in men with asthenozoospermia and serum 25-OH-D3 < 30ng/ml. Iran Clinical Trials Registry, ID: IRCT20151128025274N4, registered on 28 March 2018, URL of trial registry record: https://www.irct.ir/trial/29983.

Evaluationof the novel vaginal contraceptive agent PPCM in preclinical studies using sperm hyaluronan binding and acrosome status assays.

BackgroundPolyphenylene carboxymethylene (PPCM) sodium salt is a promising multipurpose technology for prevention of both sexually transmitted infections (STIs) and pregnancy. In preclinical studies, PPCM has demonstrated significant (1) antimicrobial activity against several important viral and bacterial pathogens and (2) contraceptive activity associated with premature acrosome loss.ObjectiveTo further evaluate a vaginal antimicrobial compound as a contraceptive agent in preclinical studies utilizing a repurposed hyaluronan binding assay (HBA).Materials and methodsSemen samples containing either neat semen or washed spermatozoa were treated with increasing concentrations of PPCM or calcium ionophore A23187 (positive control). Sperm inactivation was measured by two methods: (1) double acrosome staining (AS), and (2) a hyaluronan binding assay (HBA®). Percentage of inactivated sperm was compared between untreated control sperm and those treated with PPCM or A23187.ResultsPPCM had a significant (p < 0.05) and dose‐dependent effect on sperm inactivation in both assays, with HBA detecting a higher proportion of inactivated sperm than AS. PPCM did not affect sperm motility and exhibited equivalent responses in the neat and washed samples.DiscussionBoth HBA and AS confirmed that spermatozoa were rapidly inactivated at PPCM concentrations likely present in the vagina under actual use conditions and in a time‐frame comparable to in vivo migration of spermatozoa out of seminal plasma into cervical mucus.ConclusionPPCM vaginal gel may provide contraceptive protection as well as help with STI prevention. HBA may be a sensitive and much needed biomarker for sperm activity in future contraceptive development.

Effects of common Gram-negative pathogens causing male genitourinary-tract infections on human sperm functions

Male genitourinary tract (MGT) bacterial infections are considered responsible for 15% of male infertility, but the mechanisms underlying decreased semen quality are poorly known. We evaluated in vitro the effect of strains of Gram-negative uropathogenic species (two E.coli strains, three K. pneumoniae strains, P. aeruginosa and E. cloacae) on motility, viability, mitochondrial oxidative status, DNA fragmentation and caspase activity of human spermatozoa. All strains, except P. aeruginosa, reduced significantly sperm motility, with variable effects. Sperm Immobilizing Factor (SIF) was largely responsible for deteriorating effects on sperm motility of E. coli strains since they were completely reverted by knockout of SIF coding recX gene. Sequence alignment for RecX showed the presence of high homologous sequences in K. pneumoniae and E. cloacae but not in P. aeruginosa. These results suggest that, in addition to E.coli, other common uropathogenic Gram-negative bacteria affect sperm motility through RecX products. In addition to sperm motility, the E. coli strain ATCC 35218 also affected sperm viability, and induced caspase activity, oxidative stress and DNA fragmentation suggesting an interspecies variability in the amount and/or type of the produced spermatotoxic factors. In general, our results highlight the need for a careful evaluation of semen infections in the diagnostic process of the infertile man.

Somatic-Immune Cells Crosstalk In-The-Making of Testicular Immune Privilege.

Immunological infertility contributes significantly to the etiology of idiopathic male infertility. Shielding the spermatogenic cells from systemic immune responses is fundamental to secure normal production of spermatozoa. The body's immune system is tuned with the host self-components since the early postnatal period, while sperm first develops during puberty, thus rendering spermatogenic proteins as 'non-self' or 'antigenic.' Development of antibodies to these antigens elicits autoimmune responses affecting sperm motility, functions, and fertility. Therefore, the testes need to establish a specialized immune-privileged microenvironment to protect the allogenic germ cells by orchestration of various testicular cells and resident immune cells. This is achieved through sequestration of antigenic germ cells by blood-testis barrier and actions of various endocrine, paracrine, immune-suppressive, and immunomodulatory mechanisms. The various mechanisms are very complex and need conceptual integration to disclose the exact physiological scenario, and to facilitate detection and management of immunogenic infertility caused by disruption of testicular immune regulation. The present review aims to (a) discuss the components of testicular immune privilege; (b) explain testicular somatic and immune cell interactions in establishing and maintaining the testicular immune micro-environment; and (c) illustrate the integration of multiple mechanisms involved in the control of immune privilege of the testis.

Bi-allelic mutations in DNAH7 cause asthenozoospermia by impairing the integrality of axoneme structure.

Asthenozoospermia is the most common cause of male infertility. Dynein protein arms play a crucial role in the motility of both the cilia and flagella, and defects in these proteins generally impair the axoneme structure and cause primary ciliary dyskinesia. But relatively little is known about the influence of dynein protein arm defects on sperm flagella function. Here, we recruited 85 infertile patients with idiopathic asthenozoospermia and identified bi-allelic mutations in DNAH7 (NM_018897.3) from three patients using whole-exome sequencing. These variants are rare, highly pathogenic, and very conserved. The spermatozoa from the patients with DNAH7 bi-allelic mutations showed specific losses in the inner dynein arms. The expression of DNAH7 in the spermatozoa from the DNAH7-defective patients was significantly decreased, but these patients were able to have their children via intra-cytoplasmic sperm injection treatment. Our study is the first to demonstrate that bi-allelic mutations in DNAH7 may impair the integrality of axoneme structure, affect sperm motility, and cause asthenozoospermia in humans. These findings may extend the spectrum of etiological genes and provide new clues for the diagnosis and treatment of patients with asthenozoospermia.

P–048 Effects of bisphenol S and bisphenol F on human spermatozoa: an in vitro study

Abstract Study question Are plasticizers bisphenol S (BPS) and bisphenol F (BPF) safer alternatives to bisphenol A (BPA) for human sperm function? Summary answer Unlike BPA, the analogues BPS and BPF do not significantly affect human sperm mitochondrial functions, motility and viability What is known already The widespread distribution of BPA, along with its reputation to be an endocrine disruptor has generated concerns about possible adverse effects for human health, thus prompting the European Food Safety Authority and the Food and Drug Administration to ban the use of this chemical in many plastic products. Following such restrictions, several substitutes have been developed, with BPS and BPF representing the main replacements to BPA. While it has been demonstrated that BPA promotes oxidative damages in spermatozoa from different species, including human, the possible effects exerted by BPS and BPF on human sperm, have not yet been investigated. Study design, size, duration We explored the effect of 4 h in vitro exposure to scalar concentrations of BPS and BPF (from 10 to 400 μM), and to 400 μM BPA on sperm motility, viability, mitochondrial membrane potential (Δm) and mitochondrial generation of reactive oxygen species (ROS). In a set of experiments, the effect of a combination of both BPF (400 μM) and BPS (400 μM) on ΔΨm and mitochondrial ROS generation was also assessed. Participants/materials, setting, methods Sperm Δm was analyzed by flow cytometry with the fluorescent lipophilic cationic dye JC–1. Flow cytometric assessment of mitochondrial generation of ROS was carried out using the lipid soluble cation MitoSOX red (MSR). Sperm motility and viability were evaluated by Computer-Aided Semen Analysis (CASA) and eosin assay, respectively. Main results and the role of chance The exposure to scalar concentration of BPS did not significantly affect sperm motility and viability with respect to untreated controls. A lower, albeit not significant, sperm motility was registered in samples exposed to the highest concentrations of BPF (300 μM and 400 μM). As expected, 400 μM BPA produced a complete sperm immobilization along with a dramatically loss in sperm viability. No significant differences were observed in sperm Δm and ROS generation after exposure to scalar concentration of BPS compared to untreated controls and the trend towards lower Δm and higher mitochondrial ROS generation at the highest concentrations of BPF did not reach statistical significance. On the contrary, after 4 h exposure to 400 μM BPA a significant lower Δm and higher mitochondrial ROS generation were observed. Finally, the exposure to a combination of BPF and BPS at high concentrations (400 μM) did not significantly affect sperm Δm, or mitochondrial ROS generation, when compared to 400 μM BPA, used as positive control. Limitations, reasons for caution: The present study only evaluated BPS and BPF effects, but in daily-life people are exposed to several plasticizers containing different bisphenols at different concentrations. Therefore, adverse effects of synergic exposure to BPA analogues other than BPS and BPF, alone or in combination with BPA, cannot be ruled out. Wider implications of the findings: The analogues BPS and BPF, alone or in combination, appeared to be safer alternatives to BPA on sperm biology as they exert a neutral effect on sperm motility, viability, and mitochondrial functions even at high concentrations. These results could be useful to identify more secure plasticizer components. Trial registration number Not applicable

The characteristics of proteome and metabolome associated with contrasting sperm motility in goat seminal plasma

Sperm motility is an index tightly associated with male fertility. A close relationship between seminal plasma and sperm motility has been confirmed. This study was to assess the protein and metabolite profiles of seminal plasma obtained from adult goats with high or low sperm motility using the proteomic and metabolomic strategies. In total, 2098 proteins were found. 449 differentially abundant proteins (DAPs) were identified, and 175 DAPs were enriched in the high motility group. The obtained DAPs primarily exist in cytoplasma and extra-cellular portion. The Gene Ontology enrichment analysis demonstrated the main functional roles of these DAPs in regulating biological process, metabolic process of organic substances, cellular-metabolic process, primary-metabolic process, metabolic process of nitrogen compounds, etc. Additionally, the Kyoto-Encyclopedia of Genes and Genomes (KEGG) analysis revealed that these DAPs were primarily involved in phosphatidylinositol signaling system, salivary secretion, proteasome, apoptosis, mitophagy-animal, etc. Aided by the parallel reaction monitoring technology, the abundance changing pattern of 19 selected DAPs was consistent with that of the corresponding proteins obtained by TMT. A total of 4603 metabolites were identified in seminal plasma. 1857 differential metabolites were found between the high motility group and the low motility group, and 999 metabolites were up-regulated in the high motility group. The KEGG analysis demonstrated the primary involvement of the differential metabolites in metabolic and synthetic activities. In conclusion, we first established the proteome and metabolome databank of goat seminal plasma, detecting some proteins and metabolites which may affect sperm motility. This study will be valuable for understanding mechanisms leading to poor sperm motility.

A differential proteomic study reveals the downregulation of several plasma membrane Ca2+-binding proteins in the sperm of Atlantic salmon (Salmo salar) following cold storage

Sperm motility is a key event in fertilization, which is regulated by different factors. Among the factors that most affect sperm motility in fish are the external concentrations of Ca2+, the influence of which is highly regulated by various plasma membrane Ca2+-binding proteins. Concentrations of this cation have also been shown to be one of the most important factors affecting motility in cold-stored sperm cells. Taking into account the aforementioned aspects, we carried out a differential proteomic study on Salmo salar sperm stored at 4 °C with the aim of evaluating the protein profile on day 0 and day 14. The results of our study showed that 401 proteins were significantly downregulated (p < 0.05) on day 14, where four of them are key in the sperm motility of Salmo salar. The results of this study will allow a better understanding of the sperm activation mechanisms of Salmo salar, which will be of great importance in the design of future cold storage strategies for sperm preservation.