Sperm Fertility Research Articles

Effects of SARS-CoV-2 infection on embryological outcomes in assisted reproductive technology during the Omicron epidemic

BackgroundThe influence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection on assisted reproductive technology (ART) has received increasing attention. It has been reported that the SARS-CoV-2 RiboNucleic Acid (RNA) cannot be detected in follicular fluid and granulosa cells. However, the detection rate of SARS-CoV-2 RNA in immature oocytes and blastocysts has still unknown. Moreover, the effect of SARS-CoV-2 infection on embryological outcomes in ART during the Omicron epidemic is limited.MethodsA prospective study was performed to explore the detection rate of viral RNA in biological specimens from patients who tested positive for SARS-CoV-2 RNA and the effects of SARS-CoV-2 infection on embryological outcomes. A total of 211 patients underwent transvaginal oocyte retrieval at the Third Affiliated Hospital of Guangzhou Medical University between December 13, 2022 and December 30, 2022. Prior to transvaginal oocyte retrieval, 61 individuals tested positive for SARS-CoV-2 RNA within 24 h. Follicular fluid was preserved during oocyte retrieval. Granular cells were collected after degranulation (Intracytoplasmic sperm injection only). Immature oocytes were collected at the end of the ICSI. Unavailable blastocysts were collected on day 6 (D6). The TIANLONG SARS-CoV-2 RT-PCR-Kit was used to detect SARS-CoV-2 RNA in all samples. The COVID-19 and Non COVID-19 groups were contrasted in the following areas: fertilization rate, 2PN rate, Day 3 (D3) available embryos rate, D3 good-quality embryos rate, blastocyst formation rate, good-quality blastocyst formation rate.ResultsAll samples were negative except for an immature oocytes sample that was positive for SARS-CoV-2 viral RNA with a detection rate of 6.67%. Whether in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI), the rate of fertilization, 2PN, D3 available embryos, D3 good-quality embryos, blastocyst formation, good-quality blastocyst formation was not significantly negative different between the COVID-19 and the Non COVID-19 groups. Our findings were validated by an overview of the embryological outcome from the cycles before SARS- Cov-2 infection from the same patient.ConclusionsExcept for immature oocytes, none of the follicular fluid, granulosa cells, or blastocysts samples contained viral RNA. In addition, SARS-CoV-2 infection had no detrimental effects on the embryological outcomes of ART.

Epididymal RNase T2 contributes to astheno-teratozoospermia and intergenerational metabolic disorder through epididymosome-sperm interaction

BackgroundThe epididymis is crucial for post-testicular sperm development which is termed sperm maturation. During this process, fertilizing ability is acquired through the epididymis-sperm communication via exchange of protein and small non-coding RNAs (sncRNAs). More importantly, epididymal-derived exosomes secreted by the epididymal epithelial cells transfer sncRNAs into maturing sperm. These sncRNAs could mediate intergenerational inheritance which further influences the health of their offspring. Recently, the linkage and mechanism involved in regulating sperm function and sncRNAs during epididymal sperm maturation are increasingly gaining more and more attention.MethodsAn epididymal-specific ribonuclease T2 (RNase T2) knock-in (KI) mouse model was constructed to investigate its role in developing sperm fertilizing capability. The sperm parameters of RNase T2 KI males were evaluated and the metabolic phenotypes of their offspring were characterized. Pandora sequencing technology profiled and sequenced the sperm sncRNA expression pattern to determine the effect of epididymal RNase T2 on the expression levels of sperm sncRNAs. Furthermore, the expression levels of RNase T2 in the epididymal epithelial cells in response to environmental stress were confirmed both in vitro and in vivo.ResultsOverexpression of RNase T2 caused severe subfertility associated with astheno-teratozoospermia in mice caput epididymis, and furthermore contributed to the acquired metabolic disorders in the offspring, including hyperglycemia, hyperlipidemia, and hyperinsulinemia. Pandora sequencing showed altered profiles of sncRNAs especially rRNA-derived small RNAs (rsRNAs) and tRNA-derived small RNAs (tsRNAs) in RNase T2 KI sperm compared to control sperm. Moreover, environmental stress upregulated RNase T2 in the caput epididymis.ConclusionsThe importance was demonstrated of epididymal RNase T2 in inducing sperm maturation and intergenerational inheritance. Overexpressed RNase T2 in the caput epididymis leads to astheno-teratozoospermia and metabolic disorder in the offspring.

Preservation of rooster post-thawed sperm epigenetic modifications, fertility potential and other quality parameters in different extenders using reduced glutathione

Although rooster semen cryopreservation is an efficient procedure to spread qualified semen samples for reproductive goals, some post-thawed qualified semen samples resulted in poor fertility rate that could be related to epigenetic modifications during the cryopreservation process. This research was conducted to investigate the effect of reduced glutathione (GSH) in different cryopreservation extenders (Lake and Beltsville) on preservation of epigenetic modifications, fertility potential and other quality parameters of rooster sperm after thawing. Semen samples were collected and diluted in Lake and Beltsville extenders as follows: L-0: Lake without GSH, L-G: Lake with GSH, B-0: Beltsville without GSH, and B-G: Beltsville with GSH. After freeze-thawing process, sperm motility, membrane functionality, mitochondrial activity, acrosome integrity, viability, apoptosis status, lipid peroxidation, DNA fragmentation, ROS concentration, epigenetic modifications and fertility potential were evaluated. In results, the type of extender had no effect (P > 0.05) of post-thawed sperm quality. The treatments containing GSH presented higher (P ≤ 0.05) total motility, progressive motility, membrane functionality, mitochondrial activity, acrosome integrity, viability, DNA methylation, fertility as well as lower (P ≤ 0.05) lipid peroxidation, apoptosis, DNA fragmentation and ROS concentration than other treatments. Extender supplementation with GSH had no effect (P > 0.05) on histone methylation, histone acetylation and hatching rate. In conclusion, supplementation of rooster sperm cryopreservation extender with GSH could be an effective strategy to preserve post-thawed sperm DNA methylation, fertility and other quality parameters during reproductive programs.

Asperosaponin VI protects against spermatogenic dysfunction in mice by regulating testicular cell proliferation and sex hormone disruption

Ethnopharmacological relevanceStudies have found that the causes of male infertility are complex, and spermatogenic dysfunction accounts for 30%–65% of male infertility causes, which is the main cause of male infertility. Asperosaponin VI (ASVI) is a saponin extracted from the traditional Chinese herb Dipsacus asperoides C.Y.Cheng & T.M.Ai. However, the precise protective impact and underlying mechanism of ASVI in the therapy of spermatogenic dysfunction remain unknown. Aim of the studyTo investigate the impact of ASVI on the spermatogenic dysfunction induced by cytoxan (CTX) in mice, as well as explore any potential mechanisms. Materials and methodsPotential ASVI targets were screened using the Pharmapper and Uniprot databases, while genes related to spermatogenic dysfunction were collected from the GeneCards database. The String and Cytoscape databases were then used for PPI analysis for the common targets of ASVI and spermatogenic dysfunction. Meanwhile, the Metascape database was used for KEGG and GO analysis. In vivo experiments, spermatogenic dysfunction was induced in male mice by intraperitoneal administration of CTX (80 mg/kg). To demonstrate the possible protective effects of ASVI on reproductive organs, CTX-induced spermatogenic dysfunction mice with different dosages of ASVI (0.8, 4, 20 mg/kg per day) treatment were collected and gonad weight was detected. The testis and epididymis were detected again by H&E. To assess the impact of ASVI on fertility in male mice, we analyzed sperm quality, serum hormones, sexual behavior, and fertility. The mechanism was investigated using WB, IF, IHC, and Co-IP technology. ResultsThe ASVI exhibited interactions with 239 associated targets. Furthermore, 1555 targets associated with spermatogenic dysfunction were predicted, and further PPI analysis identified 6 key targets. Among them, the EGFR gene exhibited the highest degree of connection and was at the core of the network. Based on the GO and KEGG enrichment analysis, ASVI may affect spermatogenic dysfunction through the EGFR pathway. In vivo experiments, ASVI significantly improved CTX-induced damage to male fertility and reproductive organs, increasing sperm quality. At the same time, ASVI can resist CTX-induced testicular cell damage by increasing p-EGFR, p-ERK, PCNA, and p-Rb in the testis and by promoting the interaction of CyclinD1 with CDK4. In addition, ASVI can also regulate sex hormone disorders and protect male fertility. ConclusionsASVI improves CTX-induced spermatogenesis dysfunction by activating the EGFR signaling pathway and regulating sex hormone homeostasis, which may be a new potential protective agent for male spermatogenic dysfunction.

Testicular ACE regulates sperm metabolism and fertilization through the transcription factor PPARγ

Testis angiotensin-converting enzyme (tACE) plays a critical role in male fertility, but the mechanism is unknown. By using ACE C-domain KO (CKO) mice which lack tACE activity, we found that ATP in CKO sperm was 9.4-fold lower than WT sperm. Similarly, an ACE inhibitor (ACEi) reduced ATP production in mouse sperm by 72%. Metabolic profiling showed that tACE inactivation severely affects oxidative metabolism with decreases in several Krebs cycle intermediates including citric acid, cis-aconitic acid, NAD, α-ketoglutaric acid, succinate, and L-malic acid. We found that sperms lacking tACE activity displayed lower levels of oxidative enzymes (CISY, ODO1, MDHM, QCR2, SDHA, FUMH, CPT2, and ATPA) leading to a decreased mitochondrial respiration rate. The reduced energy production in CKO sperms leads to defects in their physiological functions including motility, acrosine activity, and fertilization in vitro and in vivo. Male mice treated with ACEi show severe impairment in reproductive capacity when mated with female mice. In contrast, an angiotensin II receptor blocker (ARB) had no effect. CKO sperms express significantly less peroxisome proliferators-activated receptor gamma (PPARγ) transcription factor, and its blockade eliminates the functional differences between CKO and WT sperms, indicating PPARγ might mediate the effects of tACE on sperm metabolism. Finally, in a cohort of human volunteers, in vitro treatment with the ramipril or a PPARγ inhibitor reduced ATP production in human sperm and hence its motility and acrosine activity. These findings may have clinical significance since millions of people take ACEi daily, including men who are reproductively active.

Molecular dynamics of JUNO-IZUMO1 complexation suggests biologically relevant mechanisms in fertilization

JUNO-IZUMO1 binding is the first known physical link created between the sperm and egg membranes in fertilization, however, how this initiates sperm-egg fusion remains elusive. As advanced structural insights will help to combat the infertility crisis, or advance fertility control, we employed all-atom Molecular Dynamics (MD) to derive dynamic structural insights that are difficult to obtain experimentally. We found that the hydrated JUNO-IZUMO1 interface is composed of a large set of short-lived non-covalent interactions. The contact interface is destabilized by strategically located point mutations, as well as by Zn2+ ions, which shift IZUMO1 into the non-binding “boomerang” conformation. We hypothesize that the latter might explain how the transient zinc spark, as released after sperm entry into the oocyte, might contribute to block polyspermy. To address a second mystery, we performed another set of simulations, as it was previously suggested that JUNO in solution is unable to bind to folate despite it belonging to the folate receptor family. MD now suggests that JUNO complexation with IZUMO1 opens up the binding pocket thereby enabling folate insertion. Our MD simulations thus provide crucial new hypotheses how the dynamics of the JUNO-IZUMO1 complex upon solvation might regulate fertility.

Infertility and Risk of Autism Spectrum Disorder in Children

Previous studies on the risk of childhood autism spectrum disorder (ASD) following fertility treatment did not account for the infertility itself or the mediating effect of obstetrical and neonatal factors. To assess the association between infertility and its treatments on the risk of ASD and the mediating effect of selected adverse pregnancy outcomes on that association. This was a population-based cohort study in Ontario, Canada. Participants were all singleton and multifetal live births at 24 or more weeks' gestation from 2006 to 2018. Data were analyzed from October 2022 to October 2023. The exposure was mode of conception, namely, (1) unassisted conception, (2) infertility without fertility treatment (ie, subfertility), (3) ovulation induction (OI) or intrauterine insemination (IUI), or (4) in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). The study outcome was a diagnosis of ASD at age 18 months or older. Cox regression models generated hazard ratios (HR) adjusted for maternal and infant characteristics. Mediation analysis further accounted for the separate effect of (1) preeclampsia, (2) cesarean birth, (3) multifetal pregnancy, (4) preterm birth at less than 37 weeks, and (5) severe neonatal morbidity. A total of 1 370 152 children (703 407 male [51.3%]) were included: 1 185 024 (86.5%) with unassisted conception, 141 180 (10.3%) with parental subfertility, 20 429 (1.5%) following OI or IUI, and 23 519 (1.7%) following IVF or ICSI. Individuals with subfertility or fertility treatment were older and resided in higher-income areas; the mean (SD) age of each group was as follows: 30.1 (5.2) years in the unassisted conception group, 33.3 (4.7) years in the subfertility group, 33.1 (4.4) years in the OI or IUI group, and 35.8 (4.9) years in the IVF or ICSI group. The incidence rate of ASD was 1.93 per 1000 person-years among children in the unassisted conception group. Relative to the latter, the adjusted HR for ASD was 1.20 (95% CI, 1.15-1.25) in the subfertility group, 1.21 (95% CI, 1.09-1.34) following OI or IUI, and 1.16 (95% CI, 1.04-1.28) after IVF or ICSI. Obstetrical and neonatal factors appeared to mediate a sizeable proportion of the aforementioned association between mode of conception and ASD risk. For example, following IVF or ICSI, the proportion mediated by cesarean birth was 29%, multifetal pregnancy was 78%, preterm birth was 50%, and severe neonatal morbidity was 25%. In this cohort study, a slightly higher risk of ASD was observed in children born to individuals with infertility, which appears partly mediated by certain obstetrical and neonatal factors. To optimize child neurodevelopment, strategies should further explore these other factors in individuals with infertility, even among those not receiving fertility treatment.

Diet Influence on Sperm Quality, Fertility, and Reproductive Behavior in Karakul of Botoșani Rams

This study aims to analyze the influence of an improved diet with vitamins and minerals (VM) on the live weight, body condition, quality of sperm, behavior, and fertility of rams. The biological material comprised two groups of rams (L1—control and L2—VM supplemented), each consisting of 15 individuals. After a complete one-year cycle, they received different dietary treatments at the beginning of the preparation for the reproduction period. Although in the onset of the mounting period (SM), no significant differences were observed for live weight (p > 0.05), providing supplemental feeding of a VM complex allowed a better capitalization of body reserves, and, consequently, the rams’ groups differed significantly by the end of mating season (FM), for live weight (+4.1%; p < 0.001) and body condition score (+15.9%; p < 0.05). Adding vitamins and minerals to the L2 diet also improved sperm color (p < 0.001), sperm concentration (+11.8%; p < 0.01), live spermatozoa (+2.6%; p < 0.001), and decreased abnormal spermatozoa proportion (−7.0%; p < 0.01). The increase in the scrotum circumference in L2 (+4.57%) suggests that VM supplements improved testosterone secretion, spermatogenesis, and ejaculate volume (+10.20%; (p < 0.001), with a positive impact (p < 0.001) on mating behavior, on the gestation installation (+11.2%) and on the number of obtained lambs (+14.0%), as well as on the key economic indicators (+13.8% incomes per ram).

Bovine nucleoprotein transitions genes and protein abundance as valuable markers of sperm motility and the correlation with fertility

Bovine nucleoprotein transitions (TNPs), specifically TNP1 and TNP2, are essential molecules in sperm nucleus rich in arginine and lysine. These molecules act in the phase between histone expulsion and before incorporation of protamine in the spermatid nucleus. Therefore, this study aimed to analyze genes and protein abundance of TNP1 and TNP2 in sperm to determine the potential as motility markers and correlation with fertility in the field. An objective evaluation method, CASA-Sperm Vision, was used to separate 22 bulls into two groups (mg-A and mg-B) based on their increasing motility. Sperm quality parameters were also examined including velocity, mitochondrial membrane potential (MMP) by the JC-1 method, head defects using William staining, and DNA fragmentation by Halomax. TNPs genes abundance was performed using the RT-qPCR method, and the protein abundance was examined with the EIA approach. The fertility rate was also analyzed based on the conception rate generated from each bull in the field, with the data obtained from iSIKHNAS. The results showed that TNPs genes and protein abundance were significantly higher (P < 0.05) in mg-A compared to mg-B, followed by various sperm quality parameters and fertility rates (P < 0.05). Positive correlations were found in TNPs genes and protein abundance with motility, velocity, MMP, and fertility (P < 0.01). Meanwhile, a negative correlation (P < 0.01) was found between head defects and DNA fragmentation. These results showed the potential of TNPs as sperm motility markers and bull fertility.

Factors determining sperm fertility

The purpose of the work is to to systematize scientific data on factors affecting sperm fertility. Sperm fertility is the ability of a sexually mature organism to reproduce offspring. It is determined by parameters such as sperm motility and the quantity of normal and pathological sperm forms in the ejaculate. The fertility of male producers is acquired in stages: spermatogenesis occurs at the beginning, then the spermatozoa mature in the epididymis. The epididymis plays a significant role in making sperm fertile. Maturation occurs due to a complex of substances produced by the epididymis, and the full maturation of spermatozoa occurs in the genital tract of the female. Thus, fructose, produced by the vesicular glands, affects the fertility of animals. Another substance found in seminal plasma is citric acid, which takes part in the acrosomal reaction. Fertility is influenced by such factors as: the age of the animal, breed, the health of the male's reproductive system, heat stress and others. For instance, as animals age, the number of pathological sperm forms increases in the ejaculate, and their motility decreases. The same happens in cases of inflammatory diseases of the male reproductive system. Changes in the qualitative characteristics of sperm were recorded among Simmental and red breeds, as well as Holstein-Frisian and Jersey breeds. Prolonged or severe heat stress can also have a negative impact on fertility, despite the presence of compensatory mechanisms. These mechanisms are maintained by the presence of sweat glands in the scrotum, which are innervated by sympathetic nerves, as well as a reflex arc between the scrotum and the respiratory center. When the scrotum's temperature rises, respiratory rate increases, facilitating cooling. Furthermore, sperm activity can vary depending on the season. In animals with seasonal reproductive activity, sperm fertility parameters are better during the breeding season.In summary, sperm fertility is a multifactorial process, the success of which is mainly due to the health of the male reproductive system and the presence of many compensatory and adaptive mechanisms.

Polystyrene Nanoplastics in Aquatic Microenvironments Affect Sperm Metabolism and Fertilization of Mytilus galloprovincialis (Lamark, 1819).

The continuous and unregulated discharge of wastes and pollutants into the aquatic environment has required constant monitoring of the risks incurred by aquatic ecosystems. Alarmism arises from plastic pollution as larger artifacts release nanoscale fragments that can contact free-living stages such as gametes, embryos, and larvae. Specifically, the interaction between spermatozoa, released in water in externally fertilizing species, and the surrounding microenvironment is essential for successful fertilization. Activation and kinematics of movement, proper maintenance of ionic balance, and chemotactism are processes highly sensitive to even minimal perturbations caused by pollutants such as polystyrene nanoplastics. Spermatozoa of Mytilus galloprovincialis (M. galloprovincialis), an excellent ecotoxicological model, undergo structural (plasma membrane ruptures, DNA damage) and metabolic (reduced motility, fertilizing capacity) damage upon exposure to 50 nm amino-modified polystyrene nanoplastics (nPS-NH2). Nanoplastics of larger diameter (100 nm) did not affect sperm parameters. The findings highlighted the negative impact that plastic pollution, related to nanoparticle diameter and concentration, could have on sperm quality and reproductive potential of organisms, altering the equilibrium of aquatic ecosystems.

379 Impacts of Serecia Lespedeza and Black Seed Meal Dietary Supplementation on Sperm Quality and Fertilityvariables of Male Goats

Abstract Infertility is a significant issue in the livestock industry. Parasitic infestation is a significant cause of infertility in small ruminants. Sericea Lespedeza (SL) and Black seed meals (BSM) have been reported to improve sperm quality and fertility in rodents; however, little is known about their effects on livestock. Our aim was to evaluate the effects of feeding male goats with SL and BSM on their sperm quality and fertility. We conducted an 8-week pen study involving 60 mature, intact Spanish bucks divided into four groups. The animals were randomly assigned to four groups and fed diets of 60% pellets and 40% concentrate. The treatment groups were SL (n = 15), SL+ BSM (n = 15), alpha-alpha with deworming (n = 15), and alpha-alpha without deworming (naturally parasitized n = 15). At the end of the trial, scrotum circumferences (SC) were measured, and the animals were transported to a processing facility for slaughter. Testicles and epididymides were collected after slaughter for measuring their weights and lengths. Epididymal spermatozoa were retrieved and evaluated for their motility, concentration, viability, abnormalities, and membrane integrities. No significant differences were observed in SC and sperm motility between the 4 groups (ranged 56.7% to 67.3%). However, sperm cell concentrations, viable sperm, and the percentage of sperm with intact acrosome were greater (P &amp;lt; 0.05) in SL and SL+BSM groups than in the other 2 groups. Furthermore, the sperm abnormalities were less (P &amp;lt; 0.05) in SL and SL+BSM groups. In conclusion, dietary supplementation with SL and BSM significantly improved sperm viability and reduced sperm abnormalities in goats. This study assures the potential benefits of SL and BSM on the fertility of male goats.

PSV-9 Cold Exposure Impacts Dna Methylation Patterns of Genes in Cattle Sperm Involved in Early Embryonic and Bone Development

Abstract DNA methylation and other epigenetic modifications are indispensable for maintaining sperm quality, fertilization capacity, and embryonic and postnatal development. In mice, environmental factors, such as stress, nutrition, or exposure to the cold condition, have been demonstrated as factors that alter methylation marks of sperm that are passed to the subsequent generation through transgenerational inheritance and genomic imprinting. Breeding bulls from northern regions of the USA are exposed to extreme cold for 2 to 3 months during the winter; however, no single study has analyzed the consequences of this cold exposure on the methylation pattern of bull sperm, which constitutes a significant knowledge gap. Therefore, this study aimed to explore the effects of cold exposure on the overall and gene-specific methylation status in sperm. We collected semen from 5 bulls during winter seasons after cold exposure and at normal temperatures during late spring. Whole Genome Bisulfite Sequencing (WGBS) was conducted to obtain the DNA methylation profile of these semen samples and to identify unique genes that have differentially methylated regions due to cold exposure (Figure 1). Cold exposure did not change the overall methylation level between the two groups but induced 438 Differentially methylated regions (DMRs) that overlapped with promoters, introns, exons, intergenic regions, shores, and shelves of CpG island (CGI) in 186 unique genes. We also identified nine unique differentially methylated genes (DMGs) (Pax6, Macf1, Mest, Ubqln1, Smg9, Trappc9, Ctnnb1, Lsm4, Peg10) involved in embryonic development and nine unique DMGs (Prmt6, Nipal1, C21h15orf40, Slc37a3, Fam210a, Raly, Rgs3, Lmbr1, Gan) involved in osteogenesis (Table 1). DMRs were located in the promoter regions and introns of these genes which preferentially involve gene silencing and alternative splicing. Among the DMGs involved in embryonic development, Mest and Peg10 are two paternally imprinted genes where only the paternal allele expresses. Peg10, required for optimal placental growth and trophoblast proliferation, overlapped with a hypermethylated DMR in the promoter region. Mest, another paternally imprinted gene, the downregulation of which results in embryonic growth retardation, had hypermethylation in introns. Moreover, Methylation-specific PCR (MS-PCR) verified the methylation changes in the identified regions. It was the first study to investigate the effect of cold exposure on DNA methylation of cattle sperm and suggest its association with altered sperm DNA methylation profiles. Differential methylation appears to alter gene expression and affect early embryonic development, osteogenic activity, and overall offspring growth performance through the imprinting effects.

Semen Evaluation, Preparation of Bangkok Roosters, and Insemination to Indonesian Native Hens

The success of artificial insemination in chickens is very dependent on sperm quality. Sperm temperature and diluent during storage can affect sperm quality. This study evaluated the quality of Bangkok chicken sperm diluted with 5% glucose and NaCl stored at 5 ⁰C and 26 ⁰C. The research design used was completely randomized. The data obtained were analyzed using a variance. The results showed that the motility of sperm preserved for 9 hours at 26 ⁰C with infused NaCl and Glucose 5% differed significantly (P&lt; 0.05) with motility of 50±0.0% and 34±8.94%, respectively. While preservation under 5 ⁰C for 9 hours with NaCl, better than Glucose 5% (P&lt;0.05) with motility of 58.00±10.95% and 38.00±10.95%, respectively. The viability of sperm preserved at 26 ⁰C with glucose 5% extender was better than NaCl (P&lt;0.05) with a value of 58.93±1.27% and 33.43±1.27%, respectively. While preservation at 5 ⁰C of viabilities of sperm under Glucose 10% and NaCl were not significantly different (P&gt;0.05) from values of 52.57±5.15% and 48.14±8.09%, respectively. The abnormality of sperm stored at 26 ⁰C and 5 ⁰C for 9 hours with NaCl and Glucose 5% were not different (P&gt;0.05). Insemination using 100 million sperm with infused NaCl extender from 18 eggs produced 94.44% (n=17) fertility and 72.22% (n=13) hatchability. While insemination using infuse glucose 5% extender, 11 eggs produced 63.64% (n=7) fertility and 54.54% (n=6) hatchability. The infusing NaCl extender produced better sperm quality, fertility, and hatchability than glucose 5% at 5 ⁰C.

Expression of sperm microRNAs related to bull fertility: A systematic review

In this study we proposed to address the following question: “Are there differentially expressed sperm microRNAs related to fertility in bulls?”. A systematic review of scientific literature until November 2022 was performed, in accordance with PRISMA guidelines. The main outcome was differentially expressed sperm microRNA from bulls with low versus high fertility profiles identified by using different methods such as field fertility evaluation and sperm laboratory analysis. Were identified 786 documents, of which 13 were selected for qualitative analysis. A total of 182 unique differentially expressed miRNAs were identified, among these, 49 miRNAs were found in common between at least two studies. It is believed that from these 49 miRNAs, it is possible that miRNAs such as miR-10a, −10b, −103, −15b, −122, −125b, −126-5p, −151-5p, −193a-5p, −196a, −27a-5p and -99b could be potential universal biomarkers to assess the reproductive potential of males.

Comparisons of conventional in vitro fertilization versus intracytoplasmic sperm injection in women with thyroid autoimmunity and non-male factor infertility, a propensity score matching analysis

The aim of the study is to compare the outcomes between the insemination methods of conventional in vitro fertilization and intracytoplasmic sperm injection in infertile women with thyroid autoimmunity and non-male factor infertility. This was a retrospective cohort study which included women with thyroid autoimmunity and non-male factor infertility. Reproductive outcomes such as embryo development parameters and clinical outcomes were compared between the two groups. The propensity score matching was applied to balance the general characteristics with significant differences between the two groups. Generalized estimating equations were used to explore the impact of ICSI on the embryo development potential of the inseminated oocytes. Sensitivity analysis using E-values was used to account for unknown confounders. After 1:2 propensity score matching, the general characteristics were all comparable. The good cleavage embryo rate, blastocyst utilization rate, and good blastocyst rate were significantly lower in the intracytoplasmic sperm injection group than those in the conventional in vitro fertilization group. After controlling for the confounding factors, intracytoplasmic sperm injection was significantly negatively associated with development of usable blastocysts and good blastocysts, while showed no impact on fertilized oocytes, usable cleavage embryos and good cleavage embryos. Although limited by the limited sample size, there were comparable clinical and obstetrical outcomes between conventional in vitro fertilization and intracytoplasmic sperm injection groups. Intracytoplasmic sperm injection neither improved the embryo development potential nor increased the clinical pregnancy and live birth rates compared to conventional in vitro fertilization in the studied population. Prospective studies that randomly divide the studied population in two the two groups and compare the reproductive outcomes are warranted.

Small Non-Coding RNAs in Male Reproduction.

Male reproductive functions are strictly regulated in order to maintain sperm production and fertility. All processes are controlled by precise regulation of gene expression, which creates specific gene expression programs for different developmental stages and cell types, and forms the functional basis for the reproductive system. Small non-coding RNAs (sncRNAs) are involved in gene regulation by targeting mRNAs for translational repression and degradation through complementary base pairing to recognize their targets. This review article summarizes the current knowledge on the function of different classes of sncRNAs, in particular microRNAs (miRNAs) and PIWI-interacting RNAs (piRNAs), during male germ cell differentiation, with the focus on sncRNAs expressed in the germline. Although transcriptionally inactive, mature spermatozoa contain a complex population of sncRNAs, and we also discuss the recently identified role of sperm sncRNAs in the intergenerational transmission of epigenetic information on father's environmental and lifestyle exposures to offspring. Finally, we summarize the current information on the utility of sncRNAs as potential biomarkers of infertility that may aid in the diagnosis and prediction of outcomes of medically assisted reproduction.

Mitopherogenesis, a form of mitochondria-specific ectocytosis, regulates sperm mitochondrial quantity and fertility.

Mitochondrial export into the extracellular space is emerging as a fundamental cellular process implicated in diverse physiological activities. Although a few studies have shed light on the process of discarding damaged mitochondria, how mitochondria are exported and the functions of mitochondrial release remain largely unclear. Here we describe mitopherogenesis, a formerly unknown process that specifically secretes mitochondria through a unique extracellular vesicle termed a 'mitopher'. We observed that during sperm development in male Caenorhabditis elegans, healthy mitochondria are exported out of the spermatids through mitopherogenesis and each of the generated mitophers harbours only one mitochondrion. In mitopherogenesis, the plasma membrane first forms mitochondrion-embedding outward buds, which then promptly bud off and thereby result in the generation of mitophers. Mechanistically, extracellular protease signalling in the testis triggers mitopher formation from spermatids, which is partially mediated by the tyrosine kinase SPE-8. Moreover, mitopherogenesis requires normal microfilament dynamics, whereas myosin VI antagonizes mitopher generation. Strikingly, our three-dimensional electron microscopy analyses indicate that mitochondrial quantity requires precise modulation during sperm development, which is critically mediated by mitopherogenesis. Inhibition of mitopherogenesis causes accumulation of mitochondria in sperm, which may lead to sperm motility and fertility defects. Our findings identify mitopherogenesis as a previously undescribed process for mitochondria-specific ectocytosis, which may represent a fundamental branch of mechanisms underlying mitochondrial quantity control to regulate cell functions during development.

Comparing reproductive outcomes between conventional in vitro fertilization and nonindicated intracytoplasmic sperm injection in autologous embryo transfer cycles: a Society for Assisted Reproductive Technology Clinic Outcome Reporting System Study

ObjectiveTo compare clinical outcomes between non-indicated ICSI and conventional insemination. DesignAutologous cycles performed at SART member clinics from 2014-2017 were identified, excluding frozen oocyte cycles. Outcomes conventional insemination (IVF) or non-indicated ICSI were analyzed separately for fresh, frozen-thawed pre-implantation genetic testing (PGT) and frozen-thawed non-PGT cycles. Subjects187,520 patients underwent 318,930 cycles, 57,516 (18.0%) using conventional IVF and 261,414 ICSI (82.0%). ExposureICSI, with or without indications [male factor, prior fertilization failure or any PGT (2012 recommendations)/single-gene PGT (2020 recommendations)]. Main Outcome MeasuresOdds ratios (ORs) for live birth rates and clinical pregnancy rates were calculated after multivariable adjustment for maternal age, BMI, infertility etiologies, prior IVF births and number oocytes retrieved. ResultsICSI was indicated in 151,627 (58.0%) of cycles according to 2012 ASRM Practice Committee recommendations, and 108,895 (41.7%) according to 2020 recommendations.In multivariable models, non-indicated ICSI among fresh cycles was associated with reduced odds of completing a blastocyst-stage transfer (OR 0.72, 95% CI (0.7, 0.75), p<0.001), resulting in reduced odds of live birth (OR 0.80, 95% CI (0.78,0.83), p<0.001). Among completed fresh transfers, clinical pregnancy and live birth rates were comparable between non-indicated ICSI and IVF.Non-indicated ICSI in frozen-thawed cycles with PGT and without PGT was associated with comparable live birth and clinical pregnancy rates to IVF in multivariable models. ConclusionNon-indicated ICSI was associated with reduced blastocyst availability in fresh cycles compared with IVF, leading to lower live birth rates. Outcomes from completed transfers were clinically comparable.

Regional abundances of binder of sperm (BSP) proteins are negatively associated with the quality of frozen-thawed bovine spermatozoa.

The localization and abundance of the sperm BSP proteins correlate with in vitro fertility in domestic bulls used in artificial insemination service. Binder of sperm (BSP) proteins, secreted mainly by the accessory sex glands, are the major protein family present in bovine seminal plasma and on the sperm surface after ejaculation. In vivo, BSP proteins facilitate sperm capacitation and sperm reservoir formation; however, their impact on sperm function within the in vitro systems is less clear. Therefore, this biomarker-based study aimed to characterize the localization and abundance of BSP proteins from in vitro processed frozen-thawed bovine spermatozoa. Using image-based flow cytometry and Western blotting, BSP protein localization, abundance, membrane and acrosomal integrity were investigated in the supernatant (nonmotile) and pellet (motile) fractions of gradient-separated bull spermatozoa. Spermatozoa from the supernatant fraction had high enrichment of all BSP proteins investigated (BSP1, BSP3, BSP5; P < 0.05) when compared to the pellet fraction. In the pellet fraction, BSP1 and BSP3 bound predominately to the acrosomal region, whereas BSP5 had a high affinity for the midpiece. However, in the supernatant fraction, BSP proteins predominately coated the entire sperm surface resulting in the loss of regional specificity. High BSP protein abundance in the spermatozoa also correlated with acrosome and membrane damage. Whereas a high abundance of BSP5 correlated with low embryo cleavage rates, high abundance of BSP1 on the sperm head coincided with a high blastocyst rate. Therefore, changes in the quantity and localization of specific BSP proteins could act as potential biomarkers of sperm quality and fertility.