Affect Sperm Motility Research Articles

Antifertility Effect of Polyaniline with Iron α-Fe2O3 on Epididymal Sperm of Mice

Chemical polymerization produces polyaniline in two concentrations (0.02,0.05g) at room temperature. The reaction of polymerization was exothermal in nature. The characterization peak was observed in the FTIR range between 1470 and 1560cm-1which defined benzenoid and quinoid ring. To find out the effects Fe2O3, this investigation was carried out with two weights of Fe2O3 (0.05 and 0.02 g). In the experiment part, 32 mice were separated into eight groups, as indicated. The experiment lasted for 35 days. The results showed a significant (P < 0.05) decrease in sperm motility, concentration, percentage of live sperm and testosterone concentration with a significant (P < 0.05) increase in the percentage of abnormal sperm in group treated with PANI-Fe2O3-Cys without magnetic field in comparison with control group. The results also showed a significant (P < 0.05) increase in sperm motility, Relative sperm count of live sperm and testosterone concentration, with a significant (P < 0.05) decrease in the percentage of abnormal sperm in group treated with PANI-Cys and PANI-Fe2O3-Cys exposed to magnetic field in comparison with the samples without magnetic field. The group treated with PANI alone showed an improvement in all parameters studied except the percentage of abnormal sperms which showed a significant (P < 0.05) decrease. Using the alternating magnetic field, it was found that it has a obvious effect on the work of the material PANI- Fe2O3-Cys and PAN-Cys. Thus, it affects sperm motility

Microbiome–Metabolome Reveals the Contribution of the Gut–Testis Axis to Sperm Motility in Sheep (Ovis aries)

A close association exists among testicular function, gut microbiota regulation, and organismal metabolism. In this study, serum and seminal plasma metabolomes, and the rumen microbiome of sheep with significant differences in sperm viability, were explored. Serum and seminal plasma metabolomes differed significantly between high-motility (HM) and low-motility (LM) groups of sheep, and 39 differential metabolites closely related to sperm motility in sheep were found in seminal plasma metabolomes, while 35 were found in serum samples. A 16S rRNA sequence analysis showed that the relative abundance of HM and LM rumen microorganisms, such as Ruminococcus and Quinella, was significantly higher in the HM group, whereas genera such as Rikenellaceae_RC9_gut_group and Lactobacillus were enriched in the mid-LM group. Serum hormone assays revealed that serum follicle-stimulating hormone (FSH) and MT levels were significantly lower in the LM group than in the HM group, whereas serum glucocorticoid (GC) levels were higher in the LM group than in the HM group, and they all affected sperm motility in sheep. Ruminococcus and other rumen microorganisms were positively correlated with sperm motility, whereas Lactobacillus was negatively correlated with FSH and GCs levels. Our findings suggest that rumen microbial activity can influence the host metabolism and hormone levels associated with fertility in sheep.

Analytical validation of five diagnostic tests for the detection of polymorphonuclear cells in stallion semen.

The objectives of this study were to evaluate the ability of five diagnostic tests to detect polymorphonuclear cells (PMNs) in stallion semen, and to determine the concentration of PMNs that affects sperm motility. We hypothesized that all tests have diagnostic value, and even low concentrations of PMNs affect motility. One ejaculate was obtained from six stallions. Aliquots of 50×106 purified sperm were incubated, in triplicate, with six concentrations of purified PMNs: 1) no PMNs, 2) 0.25×106 PMN/ml, 3) 0.5×106 PMN/ml, 4) 2.5×106 PMN/ml, 5) 5×106 PMN/ml, 6) 10×106 PMN/ml. The PMNs were quantified using a hemacytometer, cytology, a leucocyte esterase dipstick test (LEDT), a peroxidase test, and CD13 immunolabeling. Sperm motility was evaluated after 4 h at 38 °C. The number of leucocytes detected with the LEDT differed among treatments (P<0.0001), from negative results in control samples to moderate or large numbers in the samples with the highest PMN concentration. The hemacytometer count and CD13 immunostaining detected differences with the control treatment at the lowest PMN concentration (2.5×106 PMN/ml; P<0.001). Sperm motion was lower in samples with ≥5×106 PMN/ml (P<0.0001). Thus, a sample was considered leucospermic if it contained ≥5×106 PMN/ml. The LEDT had the best sensitivity (100%), followed by cytology (78%), peroxidase test (60%), CD13 immunostaining (56%) and hemacytometer count (47%). The LEDT had the lowest specificity (65%), which was 95% for all other tests. In conclusion, the LEDT was a simple, economic and sensitive stall-side test to screen semen for presence of PMNs. Because of the lower specificity, positive LEDT results should be confirmed with the identification of peroxidase-positive cells or CD13-positive cells.

The influence of preincubation time of prepared sperm before IVF on fertilization, embryo developmental competence and the reproductive outcomes.

It has been provided that if incubation time of prepared sperm can affect sperm motility and DNA fragment, but little is known about the influence of sperm preincubation time (SI) on the sperm's fertilizing ability, subsequent embryonic development and pregnancy outcomes in in vitro fertilization (IVF). The aim of this study was to explore the association of SI with fertilization rate, embryo development and clinical outcomes in IVF, further, to find an optimal preincubation time for prepared sperm before insemination in IVF. This retrospective cohort study included a total of 1453 infertile couples undergoing IVF in our center performed from January 2016 to January 2019. Sperm were preincubated at 37℃ 6% CO₂ for different times before insemination. Preincubation time associated with fertilization rate (FR), 2PN rate, D3 good quality embryo rate, fresh embryo implantation rate (IR), blastocyst formation rate, cumulative pregnancy rate (CPR), cumulative ongoing pregnancy rate (COPR), cumulative live birth rate (CLBR), newborn health and gender ratio were analyzed by chi-square analysis. FR and 2PN rate of SI more than four hours SI groups (> 4 h SI group) decreased significantly compared with other SI groups (p < 0.01). There were no significant differences of the D3 high quality embryo rate among five SI groups. The blastocyst formation rate of > 4 h SI group was significantly lower than that of 2-3 h SI group (45.5% vs 56.1%, p < 0.05); and 1-2 h SI group also had significant difference with 2-3 h and 3-4 h SI group (48.9% vs 56.1% and 54.6%, p < 0.05). There were a significant decrease of fresh IR and CPR in ≤1 h SI group compared with 1-2 h SI group (19.6% vs. 38.0%, p < 0.05; 62.7% vs 73.7%, p < 0.05); ≤ 1 h SI group also have the lowest CLBR (45.6%), it had statistic differences with 1-2 SI group and 3-4 SI group (45.6% vs 63.2%, p < 0.01; 45.6% vs 61.2%, p < 0.05). The sperm preincubated time at 37℃ 6% CO₂ before insemination could influence sperm fertilizing ability, blastocyst formation, embryo implantation and CLBR in IVF cycles. The best time for prepared sperm preincubation at 37℃ is one to four hours before insemination in IVF.

Quantitative proteomics of sperm tail in asthenozoospermic patients: exploring the molecular pathways affecting sperm motility.

Asthenozoospermia, characterized by low sperm motility, is one of the most common causes of male infertility. While many intrinsic and extrinsic factors are involved in the etiology of asthenozoospermia, the molecular basis of this condition remains unclear. Since sperm motility results from a complex flagellar structure, an in-depth proteomic analysis of the sperm tail can uncover mechanisms underlying asthenozoospermia. This study quantified the proteomic profile of 40 asthenozoospermic sperm tails and 40 controls using TMT-LC-MS/MS. Overall, 2140 proteins were identified and quantified where 156 proteins have not been described earlier in sperm tail. There were 409 differentially expressed proteins (250 upregulated and 159 downregulated) in asthenozoospermia which by far is the highest number reported earlier. Further, bioinformatics analysis revealed several biological processes, including mitochondrial-related energy production, oxidative phosphorylation (OXPHOS), citric acid cycle (CAC), cytoskeleton, stress response, and protein metabolism altered in asthenozoospermic sperm tail samples. Collectively, our findings reveal the importance of mitochondrial energy production and induced stress response as potential mechanisms involved in the loss of sperm motility in asthenozoospermia.

Diluent pH affects sperm motility via GSK3 α/β-hexokinase pathway for the efficient enrichment of X-sperm to increase the female kids rate of dairy goats

Dairy goats are the goats bred with the ability to produce large quantities of milk, and the increase of the female kid rate of breeding dairy goats is beneficial for milk production and economic benefits of dairy goat farms. Our previous study revealed that regulating the pH of dairy goat semen diluent to 6.2 or 7.4 respectively, the proportion of X chromosome bearing sperm (X-sperm) in the up and down layers of the tube after incubation was significantly higher than that of Y chromosome bearing sperm (Y-sperm) i.e. enriched X-sperm. In this study, fresh dairy goat semen collected in different seasons was diluted in different pH solutions to calculate the number and rate of X-sperm and to measure the functional parameters of enriched sperm. The artificial insemination experiments were performed with enriched X-sperm. The mechanisms of regulating the pH of diluent affecting sperm enrichment were further studied. The results showed that the proportion of enriched X-sperm in pH 6.2 and 7.4 diluents of sperm collected in different seasons showed no significantly different, but were significantly higher than that of the control group (pH 6.8). The in vitro functional parameters of X-sperm enriched in pH 6.2 and 7.4 diluent solution were not significantly different from those of the control group (P > 0.05). After artificial insemination with X-sperm enriched in pH7.4 diluent, the proportion of female offspring was significantly higher than that of the control group. It was found that the regulating pH of the diluent affected sperm mitochondrial activity and glucose uptake capacity via phosphorylating NF-κB and GSK3α/β proteins. The motility activity of X-sperm was enhanced under acidic conditions and weakened under alkaline conditions, which was conducive to the effective enrichment of X-sperm. This study demonstrated that the number and proportion of X-sperm enriched using pH 7.4 diluent were elevated, and the proportion of female kids was increased. This technology can be used for the reproduction and production of dairy goats in farms at large scales.

Decrease of air pollution during lockdown in Tuscany (Italy): An effect on sperm DNA fragmentation?

In March 2020, the Italian government imposed a national lockdown which was almost completely removed in June 2020. Due to the abrupt stop of human activities, emissions of air pollutants decreased. Air pollution is an environmental risk factor for noncommunicable disease and mortality. Emerging evidence also suggests a role in male infertility. In this study, we compared sperm DNA fragmentation (sDF) levels and conventional semen parameters between subjects undergoing sDF determination and routine semen analysis in a single Italian centre, during about 6 months before (N = 119) and after lockdown (N = 105). After lockdown, we found an improvement of sperm progressive motility (48.00[38.50-58.00]% vs. 42.00[33.00-53.00]%) and sDF levels (as total: 24.79[18.33-33.97]% vs. 35.02[25.04-45.73]%, p < .001; brighter: 14.02[10.69-17.93]% vs 18.54[13.58-25.82]%, p < .001 and dimmer sDF: 9.24[5.64-15.78]% vs. 12.24[8.08-19.10]%, p < .01), mirrored by a decrease of leukocyte semen concentration (p < .01). The improvement of sperm motility and DNA quality was maintained after adjusting for leukocyte concentration and several conditions known to affect sperm motility and/or sDF levels. With a significant decrease in air pollution observed in Tuscany during and after lockdown, associated improvement in sperm motility and DNA quality in patients referred to the infertility clinic is suggestive of the potential role of air pollution in male infertility.

Effect of type and concentration of cryoprotectant on the motility, viability, and fertility of climbing perch Anabas testudineus Bloch, 1792 (Pisces: Anabantidae) sperm

The climbing perch, Anabas testudineus is a freshwater fish that has economic value in Indonesia. It is cultured in the country, but the breeding technology, specifically sperm storage, is not well developed. Sperm cryopreservation is one of the preservation methods that need to be developed to support fish breeding technology. The type of cryoprotectants and its concentration are species-dependent and determines the success of this approach. Therefore, this study is aimed at determining the optimal type and concentration of cryoprotectant for sperm cryopreservation of A. testudineus. Four separate study series were performed, each of which evaluated one type of cryoprotectant at five concentration levels. The cryoprotectants used were DMSO, methanol, glycerol, and ethanol, and the tested concentrations were 0%, 5%, 10%, 15%, and 20%, which were combined with 5% egg yolks. Each treatment was conducted with three replications. The results showed that the type of cryoprotectant and its concentration significantly affected sperm motility, viability, and fertility of climbing perch (P < 0.05). The best outcome was obtained in DMSO, and methanol at a concentration of 10%, glycerol at 5%, and ethanol at 15%. However, the highest motility, viability, and fertility values were observed at 10% DMSO, indicating it is the best type and concentration for sperm cryopreservation of climbing perch A. testudineus.

Comparative transcriptome analysis identified crucial genes and pathways affecting sperm motility in the reproductive tract of drakes with different libido

Libido can affect the semen quality of male, and the sperm motility in semen quality parameters is a reliable index to evaluate the fertility of male. In drakes, the sperm motility is gradually acquired in testis, epididymis, and spermaduct. However, the relationship between libido and sperm motility in drakes has not been reported and the mechanisms of testis, epididymis, and spermaduct regulating the sperm motility of drakes are unclear. Therefore, the purpose of the present study was to compare the semen quality of drakes with libido level 4 (LL4) and libido level 5 (LL5), and tried to identify the mechanisms regulating the sperm motility in drakes by performing RNA-seq in testis, epididymis, and spermaduct. Phenotypically, the sperm motility of drakes (P < 0.01), weight of testis (P < 0.05), and organ index of epididymis (P < 0.05) in the LL5 group were significantly better than those in LL4 group. Moreover, compared with the LL4 group, the ductal square of seminiferous tubule (ST) in testis was significantly bigger in the LL5 group (P < 0.05), and the seminiferous epithelial thickness (P < 0.01) of ST in testis and lumenal diameter (P < 0.05) of ductuli conjugentes/dutus epididymidis in epididymis were significantly longer in the LL5 group. In transcriptional regulation, in addition to KEGG pathways related to metabolism and oxidative phosphorylation, lots of KEGG pathways associated with immunity, proliferation, and signaling were also significantly enriched in testis, epididymis, and spermaduct, respectively. Furthermore, through the integrated analysis of coexpression network and protein-protein interaction network, 3 genes (including COL11A1, COL14A1, and C3AR1) involved in protein digestion and absorption pathway and Staphylococcus aureus infection pathway were identified in testis, 2 genes (including BUB1B and ESPL1) involved in cell cycle pathway were identified in epididymis, and 13 genes (including DNAH1, DNAH3, DNAH7, DNAH10, DNAH12, DNAI1, DNAI2, DNALI1, NTF3, ITGA1, TLR2, RELN, and PAK1) involved in Huntington disease pathway and PI3K-Akt signaling pathway were identified in spermaduct. These genes could play crucial roles in the sperm motility of drakes with different libido, and all data the present study obtained will provide new insights into the molecular mechanisms regulating sperm motility of drakes.

The Effects Of Purple Grape Juice (Vitis Vinifera) On Spermatozoa Motility Of Male Wistar Rats (Rattus Norvegicus) Exposed To Electric Cigarette Smoke

Background: Infertility in men can be caused by decreased motility of spermatozoa. Electric cigarette smoke containing nicotine can produce free radicals that cause a decrease in sperm motility. Purple grape juice which contains flavonoids can act as antioxidants that can increase the motility of spermatozoa. Objective: To prove the effect of administering purple grape juice on sperm motility of male Wistar rats exposed to electric cigarette smoke.Methods: This research is an experimental study with a post-test-only control group design. Sample consisted of 28 male Wistar rats divided randomly into 4 groups, namely negative control, positive control (given electric cigarette smoke), treatment one (given electric cigarette smoke + purple grape juice 3 mL/day), and treatment two (given electric cigarette smoke + purple grape juice 5 mL/day). The adaptation was carried out for 1 week, followed by treatment for 4 weeks, then the sperm of Wistar rats were collected and the sperm motility of Wistar rats was examined. Statistical test used Saphiro Wilk and Kruskal Wallis test.Results: Progressive motility rate of K(-), K(+), P1, P2 groups consecutively: 58.33 ± 7,52; 26,66 ± 8,16; 31,66 ± 7,52; 43,33 ± 12,11. Non progressive motility rate of K(-), K(+), P1, P2 groups consecutively: 26,66 ± 10,32; 55 ± 8,36; 43,33 ± 12,11; 35 ± 12,24. Immotility rate of K(-), K(+), P1, P2 consecutively: 18,33 ± 9,83; 28,33 ± 14,71; 20 ± 6,32; 16,66 ± 5,16. Kruskal Wallis test found a significant difference in progressive motility (p=0.002) and non-progressive motility (p=0.005).Conclusion: Purple grape juice affects sperm motility of male wistar rats exposed to electric cigarette smoke.

Homozygous mutation in DNALI1 leads to asthenoteratozoospermia by affecting the inner dynein arms.

Asthenozoospermia is the most common cause of male infertility. Dynein protein arms play a crucial role in the motility of sperm flagella and defects in these proteins generally impair the axoneme structure and affect sperm flagella function. In this study, we performed whole exome sequencing for a cohort of 126 infertile patients with asthenozoospermia and identified homozygous DNALI1 mutation in one patient from a consanguineous family. This identified homozygous mutation was verified by Sanger sequencing. In silico analysis showed that this homozygous mutation is very rare, highly pathogenic, and very conserved. Sperm routine analysis confirmed that the motility of the spermatozoa from the patient significantly decreased. Further sperm morphology analysis showed that the spermatozoa from the patient exhibited multiple flagella morphological defects and a specific loss in the inner dynein arms. Fortunately, the patient was able to have his child via intracytoplasmic sperm injection treatment. Our study is the first to demonstrate that homozygous DNALI1 mutation may impair the integration of axoneme structure, affect sperm motility and cause asthenoteratozoospermia in human beings.

Integrating sperm cell transcriptome and seminal plasma metabolome to analyze the molecular regulatory mechanism of sperm motility in Holstein stud bulls.

Considering that artificial insemination is the most widely used assisted reproductive technique in the dairy industry, the semen quality of bulls is very important for selecting excellent stud bulls. Sperm motility is one of the important traits of semen quality, and related genes may be regulated by environmental factors. Seminal plasma can affect sperm cell transcriptome and further affect sperm motility through exosome or other processes. However, the molecular regulation mechanism of bull sperm motility has not been studied by combining the sperm cell transcriptome with seminal plasma metabolome. The number of motile sperm per ejaculate (NMSPE) is an integrated indicator for assessing sperm motility in stud bulls. In the present study, we selected 7 bulls with higher NMSPE (5,698.55 million +/- 945.40 million) as group H and 7 bulls with lower NMSPE (2,279.76 million +/- 1,305.69 million) as group L from 53 Holstein stud bulls. The differentially expressed genes (DEGs) in sperm cells were evaluated between the two groups (H vs. L). We conducted gene co-expression network analysis (WGCNA) on H and L groups of bulls, as well as two monozygotic twin Holstein bulls with different NMSPE values, to screen candidate genes for NMSPE. The regulatory effect of seminal plasma metabolome on the candidate genes of NMSPE was also investigated. A total of 1,099 DEGs were identified in the sperm cells of H and L groups. These DEGs were primarily concentrated in energy metabolism and sperm cell transcription. The significantly enriched Kyoto encyclopedia of genes and genomes (KEGG) pathways of the 57 differential metabolites were the aminoacyl-tRNA biosynthesis pathway and vitamin B6 metabolism pathway. Our study discovered 14 genes as the potential candidate markers for sperm motility, including FBXO39. We observed a broad correlation between transcriptome of sperm cells and seminal plasma metabolome, such as three metabolites, namely, mesaconic acid, 2-coumaric acid, and 4-formylaminoantipyrine, might regulate FBXO39 expression through potential pathways. The genes related to seminal plasma metabolites expressed in sperm cells are not only located near the quantitative trait loci of reproductive traits, but also enriched in the genome-wide association study signal of sire conception rate. Collectively, this study was the first to investigate the interplays among transcriptome of sperm cells and seminal plasma metabolome from Holstein stud bulls with different sperm motility.

Diagnosis of leucospermia in the stallion

The lack of standardized tests and interpretative criteria makes the diagnosis of stallion leucospermia difficult. Our objectives were to evaluate the ability of six diagnostic tests to detect polymorphonuclear cells (PMNs) in stallion semen, and to determine the concentration of PMNs that affects sperm motility. We hypothesized that all tests have diagnostic value, and that low concentrations of PMNs affect sperm motility. One ejaculate was obtained from six stallions. Aliquots of 50 ×106 purified sperm from each stallion were incubated with six concentrations of purified PMNs: 1) no PMNs, 2) 0.25 ×106 PMN/ml, 3) 0.5 ×106 PMN/ml, 4) 2.5 ×106 PMN/ml, 5) 5 ×106 PMN/ml, and 6) 10 ×106 PMN/ml. Each treatment was performed in triplicate. The PMNs were quantified using a hemacytometer (phase contrast microscopy), cytology (light microscopy), a commercial leucocyte esterase test (dipstick), a commercial peroxidase test (light microscopy), and immunolabeling of the neutrophil marker CD13 (flow cytometry). Sperm motility was evaluated after 4 h at 38 °C. The frequency of results obtained with the esterase test differed among treatments (P<0.0001). Negative or trace results were observed only with ≤ 0.5 ×106 PMNs/ml. A small number was observed in intermediate samples. In samples with ≥ 5 ×106 PMN/ml, only moderate or large results were obtained. Among quantitative tests, the hemacytometer count and flow cytometry detected differences with the control treatment at the lowest PMN concentration (2.5 ×106 PMN/ml; P<0.001). Total and progressive sperm motility were lower in samples with ≥5 ×106 PMN/ml than in those with lower concentrations (P<0.0001). Thus, a sample was considered leucospermic with ≥ 5 ×106 PMN/ml. The leucocyte esterase dipstick test had the best sensitivity (100%), followed by cytology (78%), peroxidase test (60%), flow cytometry (56%) and hemacytometer count (47%). The esterase test had the lowest specificity (65%), whereas the specificity of all other tests was 95%. In conclusion, presence of ≥5 ×106 PMNs/ml was associated with impaired sperm motility. The leucocyte esterase dipstick test is a simple, economic and sensitive stall-side test to screen semen for presence of PMNs in quantities capable of affecting sperm motility. Because of the lower specificity, we recommend confirming positive esterase results with the identification of peroxidase-positive cells under light microscopy, or CD13-positive cells using flow cytometry.

Topical reinforcement of the cervical mucus barrier to sperm.

Close to half of the world's pregnancies are still unplanned, reflecting a clear unmet need in contraception. Ideally, a contraceptive would provide the high efficacy of hormonal treatments, without systemic side effects. Here, we studied topical reinforcement of the cervical mucus by chitosan mucoadhesive polymers as a form of female contraceptive. Chitosans larger than 7 kDa effectively cross-linked human ovulatory cervical mucus to prevent sperm penetration in vitro. We then demonstrated in vivo using the ewe as a model that vaginal gels containing chitosan could stop ram sperm at the entrance of the cervical canal and prevent them from reaching the uterus, whereas the same gels without chitosan did not substantially limit sperm migration. Chitosan did not affect sperm motility in vitro or in vivo, suggesting reinforcement of the mucus physical barrier as the primary mechanism of action. The chitosan formulations did not damage or irritate the ewe vaginal epithelium, in contrast to nonoxynol-9 spermicide. The demonstration that cervical mucus can be reinforced topically to create an effective barrier to sperm may therefore form the technological basis for muco-cervical barrier contraceptives with the potential to become an alternative to hormonal contraceptives.

Corrigendum to: 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 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.

Reproductive toxicity of ritonavir in male: Insight into mouse sperm capacitation

Since COVID-19 began in 2019, therapeutic agents are being developed for its treatment. Among the numerous potential therapeutic agents, ritonavir (RTV), an anti-viral agent, has recently been identified as an important element of the COVID-19 treatment. Moreover, RTV has also been applied in the drug repurposing of cancer cells. However, previous studies have shown that RTV has toxic effects on various cell types. In addition, RTV regulates AKT phosphorylation within cancer cells, and AKT is known to control sperm functions (motility, capacitation, and so on). Although deleterious effects of RTV have been reported, it is not known whether RTV has male reproduction toxicity. Therefore, in this study, we aimed to investigate the effects of RTV on sperm function and male fertility. In the present study, sperm collected from the cauda epididymis of mice were incubated with various concentrations of RTV (0, 0.1, 1, 10, and 100 μM). The expression levels of AKT, phospho-AKT (Thr308 and Ser473), and phospho-tyrosine proteins, sperm motility, motion kinematics, capacitation status, and cell viability were assessed after capacitation. The results revealed that AKT phosphorylation at Thr308 and Ser473 was significantly increased, and the levels of tyrosine-phosphorylated proteins (at approximately 25 and 100 kDa) were significantly increased in a dose-dependent manner. In addition, RTV adversely affected sperm motility, motion kinematics, and cell viability. Taken together, RTV may have negative effects on sperm function through an abnormal increase in tyrosine phosphorylation and phospho-AKT levels. Therefore, individuals taking or prescribing RTV should be aware of its reproductive toxicity.

Relation of seminal plasma trace mineral in the Arabian stallion’s semen with the semen characteristics and subsequent fertility

BackgroundSeminal plasma contains several microelements like Zn, Fe, Se, and Cu that affect sperm motility and male fertility. Biochemical evaluation of seminal plasma trace elements is important for assessing fertility and diagnosing male infertility. AimsThe present study was designed to evaluate the effect of seminal fluid trace elements on sperm parameters and fertility in Arabian horses. MethodsNinety-four ejaculates from 25 Arabian stallions (4–27 years old) were used to investigate the effect of seminal fluid trace elements on semen parameters and fertility. Data divided according to season, stallion age, and fertility of stallions. The concentrations of Zn, Fe, Se, Cu, Cr and Mo were determined using an atomic absorption spectrophotometer. Percentage stallion fertility estimated by mares that conceived on their first cycle. Data were analyzed by ANOVA using SPSS statistical software program (2013), version 22.0. ResultsThere was a significant effect of season on semen volume, pH, Fe, Se, Cu, Cr, and Mo. Stallion age had a significant effect on pH, sperm motility, concentration, total motile sperm count, sperm abnormalities, Zn, and Fe. Sperm motility was higher (P < 0.05) and sperm abnormalities were lower (P < 0.05) in group IV (>70% fertility) than in group I (infertile) and group II (<50% fertility). Sperm abnormalities were low in group IV and high in groups I and II. Seminal plasma Zn and Cu levels were higher (P < 0.05) in groups III (50_70% fertility) and IV than in group I. Fe levels were lower (P < 0.05) in group IV than in groups I, II, and III. Seminal plasma Mo concentrations were higher (P < 0.05) in group III than in group I. ConclusionsHigh seminal plasma concentrations of Zn, Se, Cu, and Mo and low Fe concentrations are associated with improved stallions' semen parameters and fertility.

Seasonal changes in sperm freezability of sex-reversed female rainbow trout

In order to obtain sex-reversed females of Salmonid fish (SRF; neomales, masculinized females) the hormonal treatment of fertilized eggs or larvae is used. As a consequence, masculinized fish possess less-developed testes without functional spermatic ducts. Therefore, the semen of sex-reversed females is generally characterized by low or highly variable quality and reflects testicular rather than ejaculated semen. The aim of our study was to evaluate the alterations in sperm freezability of sex-reversed female rainbow trout during the progression of the spawning season. Sperm concentration, seminal plasma osmolality, semen pH, gonadosomatic index (GSI) and total sperm production (TSP) were monitored in fresh semen. The sperm motility parameters, adenosine triphosphate (ATP) production and sperm viability were examined throughout the spawning season in fresh and frozen/thawed semen. The day of the spawning season did not affect sperm motility of fresh semen. During the equilibration of semen, 2–3-fold increase in the sperm motility compared to fresh semen on each day of the spawning season was recorded. A decrease in the sperm motility after cryopreservation compared to equilibrated semen on each day of the spawning season was recorded. The highest ATP concentrations in fresh semen were noted on the 21st day of the spawning season, whereas in cryopreserved semen, these values were the highest on the 16th and 21st days of the spawning season. The viability of fresh semen did not change during the spawning season; however, the highest sperm viability after cryopreservation was observed between the 11th and 21st days of the spawning season. These results should be taken into account during the planning of cryopreservation of semen of valuable SRF individuals, which will be further used for fertilization of eggs under hatchery conditions.

Application of auxin-induced protein degradation technique in&amp;lt;italic&amp;gt;Caenorhabditis&amp;lt;/italic&amp;gt;&amp;lt;italic&amp;gt; elegans&amp;lt;/italic&amp;gt; spermatogenesis

<p indent="0mm">SAC-1, a widely expressed PI4P phosphatase, is an essential PI signal regulator for <italic>Caenorhabditis</italic><italic> elegans</italic> different developmental stages. In order to study its specific functions for <italic>C</italic>.<italic> elegans</italic> spermatogenesis,<italic> </italic>the auxin-induced protein degradation (AID) method was employed to cause the temporal-spatial degradation of SAC-1 in developing sperm. Degron and GFP coded sequences were inserted into the endogenous <italic>sac-1</italic> locus to generate Degron::GFP::SAC-1-expressing <italic>C</italic>.<italic> elegans</italic> strain using the CRISPR/Cas9 gene-editing technique. Then, <italic>TIR1</italic>, encoding the plant-specific F-box protein, driven by the germline-specific promoter<italic> sun-1</italic>, was introduced to construct the AID strain. The sperm motility was measured by sperm activation imaging<italic> in vitro</italic> and fluorescently labeled male sperm localization in hermaphrodite after mating. In addition, we analyzed the fusion of membranous organelles (MOs) with the plasma membrane during sperm activation and examined worm fertility. The results showed that the degradation of SAC-1 in gonads affected sperm motility. The auxin treatment of AID-SAC-1 worms caused sperm to extend shorter pseudopod with less fused MOs. Furthermore, the fertility of AID-SAC-1 worms treated with auxin was significantly decreased. In conclusion, auxin-induced SAC-1 tissue-specific degradation was successfully achieved, and the loss of SAC-1 in gonads attenuated sperm activation and sperm motility. This study provides a new method and insight for studying the functions of more widely expressed developmental regulatory proteins in specific tissues such as germline.

Roselle Calyx Extract as an Antioxidant Supplement on Tris-Base Extender for Goat Semen

In this study, we used extract from Roselle (Hisbiscus sabdariffa L.) Calyx to determine its ability as an antioxidant supplement to improve motility, progressive score and abnormality of goat’s sperms. Pooled semen from four male goats with average of live body weight 34.4 ± 0.6 kg and body condition score 2.5 ± 0.1 were aliquot into two groups: Control group; 0 mg/ml and Treatment groups; (Treatment 1:13 mg/ml, Treatment 2: 20 mg/ml, Treatment 3: 33 mg/ml, Treatment 4: 43 mg/ml of Roselle’s extract. All the semen parameter was observed at 0 hour, 0.5 hour, 1.0 hour, 1.5 hours after semen collection. The supplementation of extender with 13 mg/ml, 20 mg/ml, 33 mg/ml, 43 mg/ml of extract from Roselle Calyx does not affect sperm motility, progressive score and morphology of goat (p&gt;0.05). However, all sperm, regardless with supplementation and without supplementation of Roselle extract were maintained in good quality (60%) up to 1.5 hours. It is quite difficult to conclude that there are no effect of Roselle extract on sperm quality due to arise possibility of factors that may contribute to inefficiency effect of Roselle extract on sperm parameter. There were high possibility that the causes are due to inefficiencies of extraction method and/or degradation of anthocyanin pigment during extraction and storage process. Unsuitable extraction method could results in low yield and unstable of anthocyanin pigment. Moreover, temperature, pH and light causes the pigment to become unstable during processing and storage leading to degradation. Thus we suggest anthocyanin content monitoring in extract Roselle Calyx are required for future study.