Number Of Normal Sperm Research Articles

Effects of oxycodone hydrochloride on reproductive functions in male wistar rats

Drug iatrogenicity is a constantly evolving phenomenon, affecting several therapeutic classes. The aim of the present study was to investigate the effects of prolonged administration of oxycodone hydrochloride, a central analgesic, on reproductive functions in male rats. Materials and methods: three groups of 5 rats each were created; distilled water 0.5 ml/100g, oxycodone hydrochloride 5 and 10 mg/kg. The different products were administered orally daily for 30 days. The variables studied were: animal behaviour, prostate, testicular and epididymal weights, spermogram parameters and hormonal biomarkers (FSH, LH and testosterone levels). Results: behaviorally and morphologically, the results obtained show that oxycodone hydrochloride at doses of 5 and 10 mg/kg causes stiffness of the animals' tails, priaprism and atrophy of the prostate, testicles and epididymis, of which intensity was dose-dependent. On spermogram parameters, oxycodone hydrochloride caused a significant decrease in sperm motility (p<0.001) and in the number of normal spermatozoa (p<0.05). Administration of oxycodone hydrochloride to rats significantly decreased testosterone levels (p<0.05) and an increase in LH levels. Conclusion: Oxycodone hydrochloride, administered to animals at doses of 5 and 10 mg/kg, causes morphological disturbances of the gonads and related structures, spermogram parameters and gonadal hormones. Keywords: oxycodone hydrochloide, spermatic and hormonal parameters, gonads, rat

ANALYSIS OF VARIANCE SPERMOGRAM INDICATORS OF MEN DIFFERENT AGE GROUPS WITH ASTENOZOOSPERMIA, TERATOZOOSPERMIA AND AZOOSPERMIA

In recent decades, a decline in male fertility due to deterioration in sperm quality hasbeen noted around the world. This is probably explained by the tendency to increase the diseasesof the male genital organs and, as a result, the increase in the percentage of male infertility.In addition, there is a deterioration of the quantitative and qualitative indicators of spermogramsin practically healthy men. The average number of sperm in the ejaculate of a healthy man hashalved over the past 50 years, and the average volume of ejaculate has decreased by one third. Itis known that the multicomponent composition of the internal male genital organs is in constantrestructuring due to age changes, functional activity and the influence of various factors. That iswhy it is important to take into account both physiological and age-related changes in a man'sability to conceive.Diagnosis of male infertility includes clinical research methods and methods of laboratory-instrumental examination. Among the latter, the most important for finding out the functionalstate of the gonads and the fertilizing ability of sperm is the study of ejaculate. The object ofour research was spermograms obtained during the examination of patients at the "AlternativeClinic" reproductive medicine clinic.The purpose of the work was using biometric analysis to the different indicators of menspermograms of younger, middle and older age groups with normozoospermia (N), asthenozoospermia(AZS), teratozoospermia (T) and azoospermia (A). The task of the research wasto analyze the main indicators of spermograms of men of different age groups in normal andpathological conditions, to conduct a one-factor and two-factor variance analysis of the influenceof the studied diseases and age factor.After analyzing the main indicators of spermograms of men of younger, middle andolder age groups with asthenozoospermia, azoospermia, and teratozoospermia, it was establishedthat the indicators that undergo the greatest deviations in the studied diseases are themobility of spermatozoa according to categories A and B, the morphology of spermatozoa, theFarris fertility index, and the activity of spermatozoa.After conducting a one-factor variance analysis, we established that the share of theinfluence of the studied diseases on the overall variability of such indicators of spermograms asthe mobility of spermatozoa according to category A and B and the morphology of spermatozoain the spermograms of men of the studied age groups is within 63–98% of the total contribution(younger age group), 60–96% (middle age group) and 75–96% (older age group). The shareof influence of unaccounted factors is within 2–40% of the total contribution. The share of theinfluence of the studied diseases in the overall variability of such indicators as the Farris index and sperm activity in the spermograms of men of all studied groups is significantly reducedand is within 22–44% of the total contribution. Instead, the influence of unaccounted factors(56–78% of the total contribution) is growing significantly. This may indicate the presence ofconcomitant diseases and other pathologies in the men who took part in the research.After conducting a two-factor variance analysis, we established that the shares of theinfluence of the studied diseases on the overall variability of such indicators of spermogramsas motility of spermatozoa according to category A and B, morphology of spermatozoa, Farrisfertility index and activity of spermatozoa are decisive and are within the range of 90.7–99.9%of the total contribution, the share of the influence of the age factor on the variability of spermogramindicators of men of different age groups is insignificant.

Efficacy of GnRH Pulses in Hypogonadism Secondary to Primary Empty Sella: Case Report.

This study aims to assess the effectiveness of pulsed gonadotropin-releasing hormone (GnRH) micropump replacement therapy in the treatment of hypogonadotropic hypogonadism (HH) caused by primary empty sella (PES).The efficacy of pulsed GnRH replacement therapy using the micropump was evaluated in a middle-aged male patient with HH who had experienced the loss of his only child. Relevant literature was also consulted to compare the differences between pulse GnRH treatment and conventional treatment in terms of the development of secondary sexual characteristics, sex hormone levels, sperm production rate, and sperm activity rate in male patient with HH.In this report, a 45-year-old male diagnosed with HH and PES presented with fatigue and decreased libido. The main characteristics included decreased follicle stimulating hormone (FSH) levels of 0.03 mIU/mL, luteinizing hormone (LH) levels of 0.02 mIU/mL, and testosterone (T) levels of 0.72 nmol/L. Magnetic resonance imaging (MRI) revealed an empty sella. Semen analysis showed a small number of normal sperm with reduced motility. During treatment with the micropump pulse GnRH, the patient experienced no side effects and showed improvements in fatigue, reduced libido, sexual urge, anxiety, and feelings of inferiority. LH, FSH, and T levels returned to normal, while sperm activity rate increased to 79.9%. Ultimately, the patient's spouse achieved a natural pregnancy.Pulsed gonadotropin delivery using the micropump demonstrates good efficacy and tolerability, and aligns more closely with the physiological rhythm of GnRH secretion in the human body.

Genetic background of semen parameters in Italian Simmental bulls

The aim of this study was to estimate genetic parameters and investigate the genomic background of scrotal circumference and semen parameters in the Italian Simmental bulls. Scrotal circumference, number of normal spermatozoa, ejaculate volume, spermatozoa motility, and total spermatozoa were measured on 622 bulls, of which 603 had genotypes for 42,141 SNP. Variance components of scrotal circumference were estimated with an animal model that included the fixed effects of birth year, animal age, and measurement method, and the random effects of day of measurement and animal. In the model for the other traits, the scrotal circumference was added as a covariate to account for its influence on the semen parameters. A genome-wide association study was carried out using the ssGBLUP-approach. Heritabilities ranged from 0.07 ± 0.05 (spermatozoa motility) to 0.50 ± 0.14 (scrotal circumference). A total of 13 SNP passed the Bonferroni correction threshold and the number of significantly associated markers ranged from 1 (ejaculate volume and spermatozoa motility) to 5 (total spermatozoa). Genes already associated with reproduction parameters were retrieved close to the significant SNP. Results of the present study gave preliminary insights about the genetic determinism of semen quality in Italian Simmental bulls. Highlights Low to moderate heritabilities were estimated for semen traits. Few markers were associated with the phenotypes, suggesting their polygenic determinism. Candidate genes already associated in literature with sperm traits were found.

Effect of freeze-dried quail egg white and yolk addition tosemen extender on viability of rooster sperm stored for 6hat 4°C.

The effect of freeze-dried quail egg white and yolk addition to basic EK extender on morphology and motility of chicken broiler breeder semen was investigated. Fresh pooled semen was divided into eight parts: fresh, undiluted (control), diluted in 1:2 ratio (v/v) with basic EK extender, EK + 200 mg/ml of egg white, EK + 100 mg/ml of egg white, EK + 50 mg/ml of egg white, EK + 100 mg/ml of egg yolk, EK + 50 mg/ml of egg yolk, EK + 25 mg/ml of egg yolk. Semen samples were evaluated 15min after dilution and after 6h of storage at 4°C. In the fresh semen, the number of live normal sperm was the highest in semen diluted with EK + 200 mg of egg white and EK + 100 mg of egg yolk, while the highest sperm motility was in the neat semen. Semen storage reduced the number of normal sperm in all analysed semen samples. In the neat semen, the number of normal sperm decreased, in relation to the fresh not-stored samples, by 36.8% (from 72.3% to 35.5%), with EK extender by 9.2%, in samples enriched with egg white, from 8.4% (EK + 200 mg) to 10.0% (EK + 100 mg), and in EK with egg yolk addition, from 1.2% (EK + 50 mg) to 10.6% (EK + 100 mg). The highest percentage of motile sperm was observed in EK extender enriched with 50 mg of egg white (77.1%) and EK + 25 mg of egg yolk (65.3%).

Two acquired mouse Y chromosome-linked genes, Prssly and Teyorf1, are dispensable for male fertility‡.

Prssly (Protease, serine-like, Chr Y) and Teyorf1 (Testis expressed, chromosome Y open reading frame 1) are two acquired single-copy genes located on the distal tip of the non-pairing short arm of the mouse Y chromosome adjacent to telomeric sequence. Both genes lack X chromosome-linked homologues and are expressed in testicular germ cells. We first performed analysis of Prssly and Teyorf1 genomic sequences and demonstrated that previously reported Prssly sequence is erroneous and the true Prssly sequence is longer and encodes a larger protein than previously estimated. We also confirmed that both genes encode pseudogenes that are not expressed in testes. Next, using CRISPR/Cas9 genome targeting, we generated Prssly and Teyorf1 knockout (KO) mice and characterized their phenotype. To create Prssly KO mice, we targeted the conserved exon 5 encoding a trypsin domain typical for serine proteases. The targeting was successful and resulted in a frame shift mutation that introduced a premature stop codon, with the Prssly KO males retaining only residual transcript expression in testes. The Teyorf1 targeting removed the entire open reading frame of the gene, which resulted in no transcript expression in KO males. Both Prssly KO and Teyorf1 KO males were fertile and had normal testis size and normal sperm number, motility, and morphology. Our findings show that Prssly and Teyorf1 transcripts with potential to encode proteins are dispensable for male fertility.

20 Trials and Tribulations of Countering Insufficient Sperm Quantity with Sperm Quality, Fertility, Synchrony and Deeper Site of Insemination

Abstract Superior tools and techniques for boar semen evaluation, semen preservation and artificial insemination (AI) have helped build industry confidence and competence to reduce sperm per female inseminated without compromising reproductive results. Over the last 30 years, the average number of sperm per AI dose has decreased from about 5 to 2 billion in the US and doses containing 1.5 billion sperm have become common in some other countries. While even more genetic lag can be eliminated by further reduction of sperm dose, the level at which fecundity and fertility loss occurs is not a constant and likely varies between different farms and production systems. In general, uterine delivered sperm doses near or lower than 1 billion tend to reduce litter size and further sperm dose reduction eventually reduces pregnancy rate. Deeper catheter deposition of semen, beyond the uterine body, does not seem to effectively compensate for a limiting quantity of sperm. The exception is that surgical injection of sperm into the oviducts requires much less sperm than intrauterine insemination. Reduction of the AI-to-ovulation interval, from 24 to 8 h for example, does not seem to ameliorate the negative effects of a limiting number of sperm when sperm quality is high. The exception may be that a shorter AI-to-ovulation interval can increase reproductive results when both sperm quantity and quality are limiting. Examples include low boar fertility, advanced semen age, or sperm quality compromised by cryopreservation or sex-sorting. A better understating of boar fertility, the interaction of sub-populations of sperm with the oviductal reservoir and their participation in fertilization is likely requisite to continued reduction of the number of sperm required per female. Technologies that predict or regulate sperm quality, fertility and ovulation could prove useful in the quest to use sperm more efficiently and add value to swine AI.

Addition of MitoTEMPO to the maturation medium improves in vitro maturation competence of bovine oocytes.

The effects of MitoTEMPO, a mitochondria-targeted antioxidant, and its non-targeted parent, TEMPO, on bovine oocyte maturation competence have not been determined so far. Hence, our study was aimed to investigate the effects of supplementing maturation medium with different concentrations of MitoTEMPO (0.00, 0.10, 1.00 and 10.00 µM) or TEMPO (0.00, 5.00, 10.00 and 15.00 mM) on in vitro maturation (IVM) and fertilization (IVF) of bovine oocytes. The oocytes after IVM and IVF were evaluated for the signs of nuclear maturation and normal fertilization. The average number of spermatozoa penetrated per oocyte and the level of intracellular reactive oxygen species (ROS) were also evaluated. The results showed that percentages of bovine oocytes reached the metaphase II stage of meiosis were significantly higher in the 1.00 µM MitoTEMPO group compared to the control group (without antioxidant supplementation). The normal fertilization rate also tended to be greater in this group than the control group. In comparison with the control group, the medium supplementation with 1.00 µM MitoTEMPO led to a significant decrease in the intracellular ROS level. The average number of spermatozoa penetrated per oocyte was not significantly different among the antioxidant-treated and the non-treated groups. The TEMPO addition to the maturation medium affected neither the rate of maturation/fertilization nor the level of intracellular ROS in bovine oocytes. Based on these results, we concluded that MitoTEMPO at a concentration of 1.00 µM had beneficial effects on the quality and fertilization potential of bovine oocytes.

The mutation c.346-1G > A in SOHLH1 impairs sperm production in the homozygous but not in the heterozygous condition.

Non-obstructive azoospermia (NOA) is an important cause of male infertility, and the genetic pathogenesis is still incompletely understood. The previous study reported that heterozygous mutation of c.346-1G > A in spermatogenesis and oogenesis specific basic helix–loop–helix 1 (SOHLH1) was identified in two NOA patients and suggested it is the pathogenic factor for NOA. However, in our research, this heterozygous mutation was confirmed in three Chinese infertile patients who suffered from teratozoospermia, but they had normal sperm number. Intriguingly, a homozygous mutation of c.346-1G > A in SOHLH1 was detected in a severe oligozoospermia (SOZ) patient, characterized with severely decreased sperm count. Notably, we unprecedently revealed that this homozygous mutation of c.346-1G > A in SOHLH1 leads to the sharp decrease in various germ cells and spermatogenesis dysfunction, which is similar to the phenotype of SOHLH1 knockout male mice. Moreover, western blotting confirmed that the homozygous mutation declined SOHLH1 protein expression. Additionally, we correlated the good prognosis of intracytoplasmic sperm injection (ICSI) in the patients carrying the mutation of c.346-1G > A in SOHLH1. Thus, we suggested that the heterozygous mutation of c.346-1G > A in SOHLH1 is responsible for teratozoospermia, and this homozygous mutation in SOHLH1 impairs spermatogenesis and further leads to the reduced sperm count, eventually causing male infertility, which unveils a new recessive-inheritance pattern of SOHLH1-associated male infertility initially.

Morphological Characteristics of the Sperm of the Peregrine Falcon (Falco peregrinus) during the Reproductive Season.

The morphological characteristics of different sperm cells (normal, abnormal, and immature) in the peregrine falcon during the reproductive season were analysed. We also classified the main sperm defects found in semen. Semen samples were collected from mature peregrine falcons via cloacal massage and stained with Diff-Quik stain. The percentages of normal, abnormal, and immature sperm cells were determined by bright-field optical microscopy. The number of normal spermatozoa were greater at the initial stage and subsequently decreased during the middle and later stages of the reproductive season (p < 0.01). In contrast, the percentage of abnormal spermatozoa increased significantly in the middle and end stages of the reproductive season (p < 0.05), whereas the proportion of immature spermatozoa remained stable during the study. Head defects represented the greatest proportion of morphological abnormalities, followed by the defects in the tail and midpiece regions. A small percentage of multiple defects and cytoplasmic droplets were also observed in the falcon spermatozoa. The findings of this study might be important for the development of future conservation protocols for falcon sperm.

Light Environment Effect in the Sperm and Ribonucleic Acid Quality and Body Weight of Male Mus musculus

BACKGROUND: The mice are categorized as the nocturnal animal meaning that most of the activity conducted in dark environment. However, the treatment of mice as the object studied in the laboratory often experiences the lack attention especially the treat to light exposure to mice during the investigation period which potentially affect the result. AIM: The present study aims to investigate the effect of light environment in the sperm quality, RNA quality, and body weight of male Mus musculus. METHODS: We compared in the mice the desired parameter onto the sample of 16 mice which kept in light and dark cages for 6 weeks. RESULTS: The results indicated that there was a significant difference in the number of sperm, concentration, and purity of RNA between the mice exposed in dark and light environment. The average number of sperm from the ones kept in the dark was found twice as much as those in the light condition. The average RNA concentration in the dark was higher than in the light place but the RNA purity in the dark was lower than the light place. However, the motility and morphology of sperm was not showing any significancy different in both conditions. CONCLUSION: The result proved that the natural light of cages for treating the mice in the laboratory influences the amount of sperm and the quality RNA. Therefore, the control of the light room in the experimental laboratory needs to be considered, especially during the further experiment which related to the quality of sperm, RNA, and body weight of mice.

Na,K-ATPase α4, and Not Na,K-ATPase α1, is the Main Contributor to Sperm Motility, But its High Ouabain Binding Affinity Site is Not Required for Male Fertility in Mice.

Mammalian sperm express two Na,K-ATPase (NKA) isoforms, Na,K-ATPase α4 (NKAα4) and Na,K-ATPase α1 (NKAα1). While NKAα4 is critical to sperm motility, the role of NKAα1 in sperm movement remains unknown. We determined this here using a genetic and pharmacological approach, modifying the affinity of NKAα1 and NKAα4 for the inhibitor ouabain to selectively block the function of each isoform. Sperm from wild-type (WT) mice (naturally containing ouabain-resistant NKAα1 and ouabain-sensitive NKAα4) and three newly generated mouse lines, expressing both NKAα1 and NKAα4 ouabain resistant (OR), ouabain sensitive (OS), and with their ouabain affinity switched (SW) were used. All mouse lines produced normal sperm numbers and were fertile. All sperm types showed NKAα isoform expression levels and activity comparable to WT, and kinetics for ouabain inhibition confirming the expected changes in ouabain affinity for each NKA isoform. Ouabain at 1μM, which only block ouabain-sensitive NKA, significantly inhibited total, progressive, and hyperactivated sperm motility in WT and OS, but had no significant effect on OR or SW sperm. Higher ouabain (1mM), which inhibits both ouabain-sensitive and ouabain-resistant NKA, had little additional effect on sperm motility in all mouse lines, including the OR and SW. A similar pattern was found for the effect of ouabain on sperm intracellular sodium ([Na+]i). These results indicate that NKAα4, but not NKAα1 is the main contributor to sperm motility and that the ouabain affinity site in NKA is not an essential requirement for male fertility.

The Effect of Coffee on The Quantity of Spermatozoa of Diabetic Wistar Rats Inducted By Aloxan

Hyperglycemia in people with diabetes mellitus causes oxidative stress which can interfere with the provision of nutrients through blood vessels to the tissues that form spermatozoa so that it interferes with the process of spermatogenesis in the testicular organs. Coffee contains caffeine and chlorogenic acid which functions as an antioxidant to improve spermatogenesis abnormalities which causes decreased number of normal spermatozoa. The purpose of this study was to determine the ability of coffee in improving the number of spermatozoa of alloxan induced wistar white rats.. Methods this study uses Post-test only laboratory experimental research with control group design. Submitting 30 rats divided into 5 groups: normal control (given aquades), negative control (induced alloxan + aquades), setting 1 (induced alloxan + coffee 0.054 gram / 200 gramBB), aid 2 (alloxan induced + coffee 0.108 gram / 200 gramBB ) and treatment 3 (alloxan + coffee induced 0.162 gram / 200 gramBB). Provision of coffee carried out for 14 days, rats terminated on the 15th day. Testicles are taken then observed and counted spermatozoa. One-way ANOVA test results show the value of p = 0.000. In the LSD test between the normal control group (aquades) and negative control (diabetic rats without treatment) had a value of p = 0.042, whereas in the treatment group 1.2.3 with a negative control had a value of p = 0.000. Provision of coffee can increase the number of spermatozoa white wistar strain alloxan induced.

V-Set and Immunoglobulin Domain-Containing 1 (VSIG1), Predominantly Expressed in Testicular Germ Cells, Is Dispensable for Spermatogenesis and Male Fertility in Mice.

Simple SummaryV-set and immunoglobulin domain-containing 1 (VSIG1) is a newly discovered member of the junctional adhesion molecule (JAM) family encoded by a gene located on the X chromosome in humans and mice. Expression of VSIG1 in normal mammalian tissues was originally reported to be highly tissue-specific and was detected in organs such as the testis. However, as little is known about its physiological function, we aimed to investigate the function of VSIG1 in mammalian spermatogenesis and fertilization.To elucidate the functional role of V-set and immunoglobulin domain-containing 1 (VSIG1) in spermatogenesis and fertilization, we knocked out (KO) VSIG1 in a mouse embryo using CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9) -mediated genome editing. Reverse transcription PCR was performed using cDNA synthesized from VSIG1 KO testis RNA. Although Western blot analysis using a specific antibody to VSIG1 confirmed VSIG1 protein defects in the KO mice, hematoxylin-eosin staining analysis was similar in the KO and wild-type mice. Additionally, computer-assisted sperm analysis and in vitro fertilization experiments were conducted to confirm the activity and fertilization ability of sperm derived from the KO mouse. Mice lacking VSIG1 were viable and had no serious developmental defects. As they got older, the KO mice showed slightly higher weight loss, male mice lacking VSIG1 had functional testes, including normal sperm number and motility, and both male and female mice lacking VSIG1 were fertile. Our results from VSIG1 KO mice suggest that VSIG1 may not play essential roles in spermatogenesis and normal testis development, function, and maintenance. VSIG1 in sperm is dispensable for spermatogenesis and male fertility in mice. As several genes are known to possess slightly different functions depending on the species, the importance and molecular mechanism of VSIG1 in tissues of other species needs further investigation.

Rapid Induction of Female-to-Male Sex Change in Adult Zebrafish by Injection of an Aromatase Inhibitor.

Previously, we examined whether aromatase inhibitor (AI) treatment induces a sex change in adult female zebrafish. A 5-month AI treatment regime resulted in the retraction of the ovaries and testis formation. Eight weeks after changing the diet to AI-free food, a large number of normal sperm were obtained. Artificial fertilization using sperm from the sex-changed females was successful. These results demonstrated that sex plasticity remains in the mature ovaries of zebrafish. However, >7 months of treatment was necessary; thus, pairing was unsuccessful. In this study, we tried to induce sex change through the injection of an AI to shorten the time course of sex change. When the AI solution was directly injected into the abdomen of zebrafish, retraction of the ovary was induced within 2 months. The natural mating of sex-changed females with normal females was successful at 3 months. Although the fertilization rate was low, juveniles resulting from these matings developed normally. We succeeded in establishing a method for inducing sex changes in adult zebrafish within 3 months. The procedure will support the study of how sexual plasticity persists in adult zebrafish following sex differentiation and the identification of undifferentiated stem cells.

Why Do We Need New Markers for Male Hypogonadism and How Seminal Proteomics Might Solve the Problem?

Male hypogonadism is "a clinical syndrome that results from failure of the testis to produce physiological concentrations of testosterone and/or a normal number of spermatozoa due to pathology at one or more levels of the hypothalamic- pituitary-testicular axis". The diagnostic protocol of male hypogonadism includes accurate medical history, physical exam, as well as hormone assays and instrumental evaluation. Basal hormonal evaluation of serum testosterone, LH, and FSH is important in the evaluation of diseases of the hypothalamus-pituitary-testis axis. Total testosterone levels < 8 nmol/l profoundly suggest the diagnosis of hypogonadism. An inadequate androgen status is moreover possible if the total testosterone levels are 8-12 nmol/L. In this "grey zone" the diagnosis of hypogonadism is debated and the appropriateness for treating these patients with testosterone should be fostered by symptoms, although often non-specific. Up to now, no markers of androgen tissue action can be used in clinical practice. The identification of markers of androgens action might be useful in supporting diagnosis, Testosterone Replacement Treatment (TRT) and clinical follow-up. The aim of this review is to analyze the main findings of recent studies in the field of discovering putative diagnostic markers of male hypogonadism in seminal plasma by proteomic techniques. The identified proteins might represent a "molecular androtest" useful as a seminal fingerprint of male hypogonadism, for the diagnosis of patients with moderate grades of testosterone reduction and in the follow-up of testosterone replacement treatment.

LYPD4, mouse homolog of a human acrosome protein, is essential for sperm fertilizing ability and male fertility†

Fertilization is one of the fundamental biological processes, but so far, we still do not have a full understanding of the underlying molecular mechanism. We have identified a human acrosome protein, LY6/PLAUR domain containing 4 (LYPD4), expressed specifically in human testes and sperm, and conserved within mammals. Mouse Lypd4, also specific to the testis and sperm, is essential for male fertility. LYPD4 protein first appeared in round spermatids during acrosome biogenesis and became part of acrosomes during spermatogenesis and in mature sperm. Lypd4 knockout mice are infertile with normal sperm number and motility. Mutant sperm, however, failed to reach oviduct during sperm migration inside the female reproductive tract, leading to fertilization failure and infertility. In addition, Lypd4 mutant sperms were unable to fertilize denuded egg via IVF (in vitro fertilization) but could fertilize eggs within intact Cumulus-Oocyte Complex, supporting an additional role in sperm-zona interaction. Out of more than five thousand spermatozoa proteins identified by mass spectrometry analysis, only a small subset of proteins (26 proteins) was changed in the absence of LYPD4, revealing a whole proteome picture of mutant sperm defective in sperm migration and sperm-zona binding. ADAM3, a key component of fertilization complex, as well as other sperm ADAM proteins are significantly reduced. We hence propose that LYPD4 plays an essential role in mammalian fertilization, and further investigation of its function and its interaction with other sperm membrane complexes may yield insights into human fertilization and novel strategy to improve IVF success.

Uncovering sperm metabolome to discover biomarkers for bull fertility

BackgroundSubfertility decreases the efficiency of the cattle industry because artificial insemination employs spermatozoa from a single bull to inseminate thousands of cows. Variation in bull fertility has been demonstrated even among those animals exhibiting normal sperm numbers, motility, and morphology. Despite advances in research, molecular and cellular mechanisms underlying the causes of low fertility in some bulls have not been fully elucidated. In this study, we investigated the metabolic profile of bull spermatozoa using non-targeted metabolomics. Statistical analysis and bioinformatic tools were employed to evaluate the metabolic profiles high and low fertility groups. Metabolic pathways associated with the sperm metabolome were also reported.ResultsA total of 22 distinct metabolites were detected in spermatozoa from bulls with high fertility (HF) or low fertility (LF) phenotype. The major metabolite classes of bovine sperm were organic acids/derivatives and fatty acids/conjugates. We demonstrated that the abundance ratios of five sperm metabolites were statistically different between HF and LF groups including gamma-aminobutyric acid (GABA), carbamate, benzoic acid, lactic acid, and palmitic acid. Metabolites with different abundances in HF and LF bulls had also VIP scores of greater than 1.5 and AUC- ROC curves of more than 80%. In addition, four metabolic pathways associated with differential metabolites namely alanine, aspartate and glutamate metabolism, β-alanine metabolism, glycolysis or gluconeogenesis, and pyruvate metabolism were also explored.ConclusionsThis is the first study aimed at ascertaining the metabolome of spermatozoa from bulls with different fertility phenotype using gas chromatography-mass spectrometry. We identified five metabolites in the two groups of sires and such molecules can be used, in the future, as key indicators of bull fertility.

Quality of fresh and cryopreserved semen and their influence on the rates of fertilization, hatching and quality of the larvae of Piaractus mesopotamicus.

In this work, the seminal parameters of P. mesopotamicus were evaluated fresh and after cryopreservation, focusing on the sperm variables that affect the rates of fertilization, hatching and post-hatching parameters such as larval survival and morphology. The semen and oocytes from the animals were collected after extrusion, and seminal quality and oocyte fertilization were analyzed. Subsequently, a portion of each semen sample was cryopreserved and, after two days, the oocytes from three new females were fertilized with cryopreserved semen from the males. The analyzes showed that progressive motility, spermatic vigor, motility duration, number of normal sperm and secondary abnormalities were higher in fresh semen than in semen after thawing (P <0.0001). Similarly, fertilization and hatching rates and the percentage of normal and abnormal larvae in fertilized oocytes were higher when fresh semen was used (P <0.0001). The cryopreservation process affected the qualitative parameters of the semen of Piaractus mesopotamicus. The primary abnormality of spermatozoa was the main variable that influenced both fertilization and hatching rates, both in fresh and thawed semen. The second most important variable that influenced, particularly, thawed semen, was the spermatic vigor.

Perubahan Kualitas Sperma Mencit (Mus musculus) Yang Terpapar Asap Rokok Elektrik

Smoking is one of the habits that can affect human health. One type of cigarette that is currently the choice is electric cigarettes. The e-cigarette content includes nicotine, propylene glycol, flavorings and glycerin. The purpose of this study was to determine the effect of exposure to e-cigarette smoke on mice strain balb / c with the parameters of the number of spermatozoa. This study used 15 male mice which were 8-10 weeks old and weighed 25-30 grams of male mice which were divided into 3 treatment groups namely control group, treatment group one and treatment group two. In the control group no exposure to cigarette smoke was carried out while in the treatment group one was exposed to cigarette smoke in mice with a dose of 0 mg nicotine and the second treatment group was exposed to e-cigarette smoke at a dose of 18 mg nicotine. Electric cigarette smoke exposure is carried out for 30 days using a suction device from a syringe. The exposure of cigarette smoke is carried out every day with successive doses of 20 times as much as 3 times with a time lag of 10 minutes (5 minutes the chamber is closed and 5 minutes of the chamber is opened). The results showed that the average number of spermatozoa in the control group was 16.816 million / ml. The average number of spermatozoa in treatment one was 10.432 million / ml and the average number of spermatozoa in treatment two was 5.234 million / ml. This shows that exposure to cigarette smoke can reduce the number of spermatozoa from the control group to the first treatment by 42.56% and the second treatment shows a decrease from the first group by 34.65%.