Sperm Fertility Research Articles

Effect of cooling rate, extender type, and storage temperature on chilled sperm quality and pregnancy rate in Kail sheep

This study aimed to evaluate how cooling rates, extender type, and storage temperatures affect sperm quality and fertility in Kail sheep. In the first experiment, pooled semen collected from 5 adult Kail rams was extended with a Tris-based extender and divided into three aliquots. The temperature was reduced from 37 °C to 5 °C in 30 min (rapid cooling), 60 min (moderate cooling), and 90 min (slow cooling). In the second experiment, pooled semen was extended with three extenders: Tris, skim milk, and sodium citrate-based, and stored at 5 °C and 12 °C for three days. In the third experiment, ewes (n = 63) were synchronized for estrus using a progestin-based protocol, and fixed-time cervical insemination (FTCI) was performed at 48 h (n = 25) and 60 h (n = 38) after the sponge removal using semen extended in Tris-based extender. Results revealed that slow cooling rates had less (P < 0.05) detrimental impact on sperm motility and kinematics when compared to both moderate and rapid cooling rates. Moreover, semen extended with Tris-based extenders had a higher (P < 0.05) percentage of motile, progressive motile, rapid progressive, and medium progressive motile sperm compared to extenders containing skim milk and sodium citrate. Storage of semen at 5 °C better preserved sperm quality compared to 12 °C. Storage duration exerted a negative influence (P < 0.05) on sperm quality across all three extender types. The overall 55.5% pregnancy rate was achieved. The season of insemination had a significant impact while the timing of insemination did not influence the pregnancy rate. In conclusion, sperm quality is better preserved when semen is stored at 5 °C and subjected to a slow cooling rate using a Tris-based extender, and the success of FTCI is contingent upon the season of insemination in Kail sheep.

Advances in microfluidic technology for sperm screening and in vitro fertilization.

About 18% of reproductive-age adults worldwide are affected by infertility. In vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) are widely used assisted reproductive technologies (ARTs) aimed at improving clinical outcomes. Efficient and noninvasive selection and isolation of highly motile sperm with intact DNA are essential for the success of IVF and ICSI and can potentially impact the therapeutic efficacy and the health of the offspring. Compared to traditional methods, microfluidic technology offers significant advantages such as low sample consumption, high efficiency, minimal damage, high integration, similar microenvironment, and high automation, providing a new platform for ARTs. Here, we review the current situation of microfluidic technology in the field of sperm motility screening and evaluation and IVF research. First, we focus on the working principle, structural design, and screening results of sperm selection microfluidic platforms. We then highlight how the multiple steps of the IVF process can be facilitated and integrated into a microfluidic chip, including oocyte capture, sperm collection and isolation, sperm sorting, fertilization, and embryo culture. Ultimately, we summarize how microfluidics can complement and optimize current sperm sorting and IVF protocols, and challenges and possible solutions are discussed.

Second ejaculation produces good quality sperm and blastocyst and decreases the rate of unexpected ICSI cycle: a propensity score-matched analysis

BackgroundSecond ejaculation can influence sperm quality which may define the first-line treatment. The purpose of this study was to evaluate the effectiveness of a second ejaculation in decreasing the unexpected intracytoplasmic sperm injection (ICSI) rate by a propensity score-matched (PSM) analysis.MethodsPatients who were projected to undergo IVF were included between January 2016 and November 2021 in this monocentric, retrospective analysis. 2782 patients included in the study, 143 and 2639 patients were non-randomized in the unexpected ICSI and IVF groups, respectively. One hundred fourteen patients with unexpected ICSI produced two semen samples on the day of ovum pick-up. After 1:4 PSM, we matched 61 patients in the second ejaculation IVF group to 238 patients in the conventional IVF group. Outcomes of sperm quality, fertilization rate, embryo quality, and pregnancy were compared.ResultsSecond ejaculation significantly improved sperm concentration, progressive motility before and after sperm swim-up, total progressive motility sperm count after swim-up, and decreased sperm DNA fragmentation (SDF). Sixty-one of 114 (53.5%) unexpected ICSI couples had enough total progressive motility sperm for IVF with the second ejaculation. There were no differences in basic clinical characteristics between couples in second ejaculation IVF and matched-conventional IVF group. For the two groups, no differences were observed in IVF outcomes. However, a significant increase in good-quality blastocyst rate was observed for second-ejaculation IVF couples. Univariate and multivariate linear regression analysis also confirmed that the second ejaculation was an independent risk factor for the good quality blastocyst rate.ConclusionSecond ejaculation could be an economical and secure alternative to get good quality sperm, and blastocyst and decrease the rate of unexpected ICSI. Multicenter studies should be conducted to confirm the potential advantages of using second ejaculation IVF in effectively reducing the rate of ICSI.

Comparison of outcomes between intracytoplasmic sperm injection and in vitro fertilization inseminations with preimplantation genetic testing for aneuploidy, analysis of Society for Assisted Reproductive Technology Clinic Outcome Reporting System data

To evaluate whether insemination via intracytoplasmic sperm injection provides any benefit over in vitro fertilization for non-male factor infertility with respect to pre-implantation genetic testing results and pregnancy outcome. Retrospective Cohort Study of the Society for Assisted Reproductive Technology database. Patients undergoing in vitro fertilization or intracytoplasmic sperm injection in non-male factor pre-implantation genetic testing for aneuploidy cycles. In vitro fertilization vs. intracytoplasmic sperm injection MAIN OUTCOME MEASURES: Primary outcomes were percentage of embryos suitable for transfer and live birth rates. Secondary outcomes included subgroup analysis for embryos suitable for transfer on cycles from patients ≥ 35 y/o vs. < 35 y/o, ≤ 6 oocytes retrieved vs. > 6 oocytes retrieved, and unexplained infertility. Additionally, gestational age at delivery and birthweight between in vitro fertilization and intracytoplasmic sperm injection were evaluated. A total of 30,446 non-male factor preimplantation genetic diagnosis for aneuploidy cycles were evaluated, of which 4867 were in vitro fertilization and 25,579 were intracytoplasmic sperm injection. Following exclusion criteria and adjustment for any necessary confounding variables, no significant differences existed in embryos suitable for transfer between in vitro fertilization and intracytoplasmic sperm injection cycles, 41.6% (40.6%, 42.6%) vs. 42.5% (42.0%, 42.9%), respectively or live birth rates, 50.1% (37.8, 62.4%) vs. 50.8% (38.5%, 62.9%), respectively. There were no significant differences in the rates of embryos suitable for transfer and live birth rates between in vitro fertilization and intracytoplasmic sperm injection in non-male factor cycles undergoing pre-implantation genetic testing for aneuploidy.

Multiple ageing effects on testicular/epididymal germ cells lead to decreased male fertility in mice

In mammals, females undergo reproductive cessation with age, whereas male fertility gradually declines but persists almost throughout life. However, the detailed effects of ageing on germ cells during and after spermatogenesis, in the testis and epididymis, respectively, remain unclear. Here we comprehensively examined the in vivo male fertility and the overall organization of the testis and epididymis with age, focusing on spermatogenesis, and sperm function and fertility, in mice. We first found that in vivo male fertility decreased with age, which is independent of mating behaviors and testosterone levels. Second, overall sperm production in aged testes was decreased; about 20% of seminiferous tubules showed abnormalities such as germ cell depletion, sperm release failure, and perturbed germ cell associations, and the remaining 80% of tubules contained lower number of germ cells because of decreased proliferation of spermatogonia. Further, the spermatozoa in aged epididymides exhibited decreased total cell numbers, abnormal morphology/structure, decreased motility, and DNA damage, resulting in low fertilizing and developmental rates. We conclude that these multiple ageing effects on germ cells lead to decreased in vivo male fertility. Our present findings are useful to better understand the basic mechanism behind the ageing effect on male fertility in mammals including humans.

Gender Control of Mouse Embryos by Activation of TLR7/8 on X Sperm via Ligands dsRNA-40 and dsRNA-DR.

Gender control technologies are promising for enhancing the production efficiency of the farm animal industry, and preventing sex-linked hereditary diseases in humans. It has been shown that the X sperm of mammalian animals specifically expresses X-chromosome-derived toll-like receptor 7/8 (TLR7/8), and the activation of TLR7/8 on the X sperm by their agonist, R848, can separate X and Y sperm via the specific inhibition of X sperm motility. The use of R848-preselected sperm for fertilization resulted in sex-ratio-skewed embryos or offspring. In this study, we aimed to investigate whether two other TLR7/8 ligands, double-stranded RNA-40 (dsRNA-40) and double-stranded RNA-DR (dsRNA-DR), are also effective in the separation of mouse X and Y sperm and the subsequent generation of gender-ratio-skewed in vitro fertilization (IVF) embryos. Our results indicated that cholesterol modification significantly enhances the transfection of dsRNA-40 and dsRNA-DR into sperm cells. dsRNA-40 and dsRNA-DR incubation with mouse sperm could separate X and Y sperm by the specific suppression of X sperm motility by decreasing its ATP level and mitochondrial activity. The use of a dsRNA-40- or dsRNA-DR-preselected upper layer of sperm, which predominantly contains high-motility Y sperm, for IVF caused a male-biased sex ratio shift in resulting embryos (with 65.90-74.93% of embryos being male). This study develops a simple new method for the efficient separation of mammalian X and Y sperm, enabling the selective production of male or female progenies.

Sperm DNA fragmentation index affect pregnancy outcomes and offspring safety in assisted reproductive technology

The role of sperm DNA fragmentation index (DFI) in investigating fertility, embryonic development, and pregnancy is of academic interest. However, there is ongoing controversy regarding the impact of DFI on pregnancy outcomes and the safety of offspring in the context of Assisted Reproductive Technology (ART). In this study, we conducted an analysis of clinical data obtained from 6330 patients who underwent in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) at the reproductive medical center of The First People's Hospital of Shangqiu and The Affiliated Hospital of Zhengzhou University. The patients was stratified into two distinct groups: IVF group and ICSI group, Within each group, patients were further classified into three subgroups. IVF: group A (< 15%) included 3123 patients, group B (15–30%) included 561 patients, and group C (≥ 30%) included 46 patients. ICSI: group A (< 15%) included 1967 patients, group B (15–30%) included 462 patients, and group C (≥ 30%) included 171 patients. Data were collected and subjected to statistical analysis. There were no significant differences in the basic characteristics among the three groups, and the sperm DFI did not significantly affect the fertilization rates, pregnancy rates, stillbirth rates and the number of birth defects. However, the incidences of miscarriage rates in IVF/ICSI groups with DFI > 30% and DFI 15–30% were significantly higher than those in IVF/ICSI groups with DFI < 15%, and the miscarriage rates in ICSI group with DFI > 30% were significantly higher than DFI 15–30% group, the smooth fitting curve shows that there is a positive correlation between miscarriage rates and sperm DFI (OR 1.095; 95% CI 1.068–1.123; P < 0.001). The birth weight of infants in the IVF/ICSI groups with DFI > 30% and DFI 15–30% exhibited a statistically significant decrease compared to those in the IVF/ICSI groups with DFI < 15%. Furthermore, the birth weight of infants in the ICSI group with DFI > 30% was lower than that of the DFI 15–30% group. The smooth fitting curve analysis demonstrates a negative association between birth weight and sperm DFI (OR 0.913; 95% CI 0.890–0.937; P < 0.001). Sperm DFI has an impact on both miscarriage rates and birth weight in assisted reproductive technology. The smooth fitting curve analysis reveals a positive correlation between miscarriage rates and DFI, while a negative correlation is observed between birth weight and DFI.

Propolis Protects GC-1spg Spermatogonial Cells against Tert-Butyl Hydroperoxide-Induced Oxidative Damage.

Propolis is a natural resin produced by honeybees with plenty of pharmacologic properties, including antioxidant activity. Oxidative stress disrupts germ cell development and sperm function, with demonstrated harmful effects on male reproduction. Several natural antioxidants have been shown to reduce oxidative damage and increase sperm fertility potential; however, little is known about the effects of propolis. This work evaluated the role of propolis in protecting spermatogonial cells from oxidative damage. Propolis' phytochemical composition and antioxidant potential were determined, and mouse GC-1spg spermatogonial cells were treated with 0.1-500 µg/mL propolis (12-48 h) in the presence or absence of an oxidant stimulus (tert-butyl hydroperoxide, TBHP, 0.005-3.6 µg/mL, 12 h). Cytotoxicity was assessed by MTT assays and proliferation by Ki-67 immunocytochemistry. Apoptosis, reactive oxygen species (ROS), and antioxidant defenses were evaluated colorimetrically. Propolis presented high phenolic and flavonoid content and moderate antioxidant activity, increasing the viability of GC-1spg cells and counteracting TBHP's effects on viability and proliferation. Additionally, propolis reduced ROS levels in GC-1spg, regardless of the presence of TBHP. Propolis decreased caspase-3 and increased glutathione peroxidase activity in TBHP-treated GC-1spg cells. The present study shows the protective action of propolis against oxidative damage in spermatogonia, opening the possibility of exploiting its benefits to male fertility.

Unravelling the epigenetic impact: Oxidative stress and its role in male infertility-associated sperm dysfunction

Male infertility is a multifactorial condition influenced by epigenetic regulation, oxidative stress, and mitochondrial dysfunction. Oxidative stress-induced damage leads to epigenetic modifications, disrupting gene expression crucial for spermatogenesis and fertilization. Paternal exposure to oxidative stress induces transgenerational epigenetic alterations, potentially impacting male fertility in offspring. Mitochondrial dysfunction impairs sperm function, while leukocytospermia exacerbates oxidative stress-related sperm dysfunction. Therefore, this review focuses on understanding these mechanisms as vital for developing preventive strategies, including targeting oxidative stress-induced epigenetic changes and implementing lifestyle modifications to prevent male infertility. This study investigates how oxidative stress affects the epigenome and sperm production, function, and fertilization. Unravelling the molecular pathways provides valuable insights that can advance our scientific understanding. Additionally, these findings have clinical implications and can help to address the significant global health issue of male infertility.

High-concentration bovine serum albumin enhances fertilization ability of cold-stored rat sperm.

Cold transport of the cauda epididymides is a useful technique for shipping laboratory rat sperm. Cold transport of rat sperm avoids potential risks of microbiological infection, animal escape or death, and animal welfare issues. Previously, we reported that a cold-storage solution containing dimethyl sulfoxide and quercetin maintained the fertility of cold-stored rat sperm. However, cold-stored rat sperm exhibited a decreased fertilization rate after 24-h storage. To recover the fertility of cold-stored sperm, we focused on the effects of bovine serum albumin (BSA), a cholesterol acceptor that induces sperm capacitation. We sought to determine the optimal concentration of BSA in fertilization medium based on the fertility of cold-stored rat sperm. High concentrations of BSA (40 mg/ml) enhanced the fertilization rate of cold-stored rat sperm and maintained sperm fertility for 144 h. Embryos derived from cold-stored and BSA-treated sperm normally developed into pups after embryo transfer. In summary, high BSA concentrations enhanced the fertility of cold-stored rat sperm and prolonged the storage period to 144 h, thereby expanding the transportable region for genetically engineered rats.

Effects of different ages on frozen semen quality and in vitro fertilization efficiency in Wannan black pigs.

According to previous studies, the quality and fertilization rate of fresh sperm from boars of different ages were significantly different. However, the difference of freeze-thaw sperm quality and fertility in boars of different ages is unclear. In this study, boars of a Chinese native breed were assigned into two groups. Each group consisted of five boars aged aged either 2-3 years (young boars = YB) or 5-6 years (aging boars = AB) A total of 60 ejaculates for each group were collected and cryopreserved. Semen quality and in vitro fertility of post-thaw sperm was evaluated. The results showed that the concentration and motility of fresh sperm collected from AB were similar to YB, but their semen volume was higher than that in YB (p < 0.05). Frozen-thawed sperm of AB had lower viability than YB, and higher abnormal rate and reactive oxygen species (ROS) levels of YB (p < 0.05). There was no effect of the age on post-thaw sperm motility and time survival. Functional assessments indicated that increasing age markedly compromises the integrity of the sperm plasma membrane and acrosome, as well as mitochondrial functionality post-thaw, albeit without affecting DNA integrity. Furthermore, increasing age of boars reduces the ability of sperm to bind to the oocyte zona pellucida after thawing, delaying the time of the first embryo cleavage after fertilization. Finally, the early developmental efficiency of in vitro fertilized embryos progressing from 4-cell to blastocyst derived from post-thaw sperm in AB significantly decreased compared to those from YB (p < 0.05). Taken together, these results suggest that increasing age in boars impairs the quality and in vitro fertility of frozen thawed sperm.

DESIGN, SYNTHESIS, SPECTRAL ANALYSIS, ANTIMICROBIAL EVALUATION AND TOXICOLOGICAL ASPECTS OF BIVALENT TIN COMPLEXES WITH MACROCYCLIC MOIETY

The novel complexes Sn(II), were synthesized of macrocyclic ligands derived from various diamine and dicarboxylic acids in the ratio of 2:2 and characterized by elemental analysis, molar conductance measurements, mass, NMR, FT-IR, and electronic spectra. The interactions of macrocyclic ligands (MLn) with SnCl2 in a 1:1 molar ratio in methanol produced the ensuing physiologically active [Sn(MLn)Cl2] type complexes, where n = 1 or 2. The complexes were initially identified using conductivity tests, molecular weight calculations, and elemental investigations. The bonding mode was determined using spectral data from the IR, 1H NMR, and 119Sn NMR. The complexes exhibit hexacoordinated octahedral geometry. The positive findings of Brine shrimp lethality of the ligands and their complexes have been discussed. The antimicrobial effects of each complex on several types of harmful fungus and bacteria have been assessed. The research focused on investigating and analyzing various aspects related to male fertility. It specifically investigated cauda epididymal spermatozoa density, sperm motility, testicular sperm density, fertility, and biological constraints of propagative tissues. The findings and discussions about these factors were presented in the study.

The influence of selection of sires according to the degree of plumage pigmentation on the autosexness of guinea fowl

The studies were carried out on cream and silver guinea fowl populations at Genefond LLC, Moscow region. The purpose of the work was to study the effect of selecting sires with contrasting down pigmentation on the accuracy of sexing guinea fowl offspring. The basis for the creation of an autosex maternal form was cream and silver guinea fowl, carriers of the silver “S” and golden “s” genes, which in preliminary experiments showed the possibility of sexing them according to the degree of plumage pigmentation. To conduct the study, two groups of guinea fowl with cream and silver colored plumage were recruited at 20 weeks of age. In each group there were 6 selection nests; males with weak pigmentation and females with strong plumage pigmentation were taken into the nest. The ratio of males to females in the nest was 1:6. Experiments have shown the absence of significant differences between cream and silver guinea fowl populations in quantitative and qualitative indicators of sperm production, fertilization and egg hatchability. The selection of producers when recruiting a breeding herd with a contrasting plumage color (males with weak, females with strong plumage pigmentation) ensured an increase in the accuracy of sexing cream guinea fowl by 5.1%, silver guinea fowl by 1.8%, while the number of nests with sexing accuracy was more than 95 %, for cream guinea fowl it was 50%, for silver guinea fowl 66.7%, respectively.

Sperm quality and in vitro fertilizing ability of boar spermatozoa stored at 4 °C versus conventional storage for 1 week.

Since boar spermatozoa show a marked deterioration in sperm quality when cooled, insemination doses are usually stored at 16-18 °C. However, maintaining this temperature during transport of semen doses is challenging, particularly during the summer months. An alternative could be to store the doses at 4 °C if cold-shock to the sperm could be prevented. The objective of this study was to evaluate boar sperm quality and fertility in in vitro fertilization after storage in AndroStar Premium at 4 °C for 1 week. Insemination doses (n = 9) in AndroStar Premium from a commercial boar semen collection station were transported to the laboratory at approximately 20 °C. At the laboratory, sperm quality evaluation and was preformed and each dose was split; half of each ejaculate was stored in a climate-controlled box at 16-18 °C, the other was slowly cooled to 4 °C. Both samples were stored for 1 week before further sperm quality evaluation and in vitro fertilization (IVF) were performed. Mean values were tested using generalized linear regression, with treatment and boar as fixed factors; p ≤ 0.05 was considered significant. Sperm membrane integrity (mean ± sem: 91 ± 0.05 and 83 ± 0.09% for 16 and 4 °C, respectively) and superoxide production (6.79 ± 2.37 and 13.54 ± 6.23% for 16 and 4 °C, respectively), were different between treatments. The DNA fragmentation index was lower in cold-stored samples than in conventionally stored samples (3.74 ± 2.25 and 7.40 ± 3.36% for 4 and 16 °C, respectively). The numbers of oocytes developing to blastocyst on Day 6 (mean ± sd: 9.0 ± 8.0 and 6.0 ± 5.0%, for storage at 16 and 4 °C, respectively) were not different between treatments. Therefore, storage of boar semen doses in AndroStar Premium at 4 °C for up to 7 days would be a viable alternative to current praxis.

Mendelian randomization analysis and validation supports MEGF9 and MLLT11 as potential targets for the treatment of varicocele and male infertility.

A growing body of research suggests a link between varicocele and male infertility (MI). However, current evidence is mainly based on retrospective studies, which are prone to interference from confounding factors and cannot establish causal relationships. Mendelian randomization (MR) studies on the causal relationship between varicocele and MI are very limited. Therefore, this study conducted a two-sample MR study to elucidate the causal effect between the two. Download the data set GSE216907 from the GEO database, and use R software to screen differential genes in normal and varicocele tissue samples. The drug targets of Bu Shen Huo Xue Prescription (BSHXP) were derived from the Herb database. All genetic datasets were obtained using publicly available summary statistics based on individuals of European ancestry from the IEU GWAS database. MR analysis was performed using MR Egger, weighted median (WM) and inverse variance weighted (IVW) methods to assess the causal relationship between exposure and outcome and to validate the findings by comprehensively evaluating the effects of pleiotropic effects and outliers. The renal vein constriction method was used to establish a pathological model of varicocele infertility. The drug was administered continuously for 60 days and the relevant indicators of the rats were observed. Obtain two therapeutic targets for varicocele through intersection analysis: MEGF9 and MLLT11, and were verified by molecular docking. MR analysis showed that MEGF9 was positively associated with MI (MR Egger, OR: 1.639, 95% CI: 1.124-2.391, P = 0.024; WM, OR: 1.235, 95% CI: 1.003-1.521, P = 0.047). MEGF9 is also positively associated with MI (IVW, OR: 1.35, 95% CI: 1.069-1.705, P = 0.012). Sensitivity analysis showed no heterogeneity and horizontal pleiotropy. The expression of MEGF9 and MLLT11 increased in the varicocele model group, while the expression decreased after treatment with low, medium, and high doses of BSHXP. In addition, the sperm number, motility, morphology, and fertility of rats in the model group were significantly lower than those in the control group (P<0.05). After BSHXP treatment, all indicators were significantly better than those of the model group (P<0.05). In conclusion, this study indirectly supports that varicocele causes MI. BSHXP inhibiting MEGF9 and MLLT11 may become a potential therapeutic target for alleviating varicocele and MI.

Assessing the efficacy of different semen extenders on post-thaw quality of Bangladeshi buffalo sperm under a manual cryopreservation protocol

Cryopreservation has been used extensively for cattle in Bangladesh, albeit no study was conducted on the cryopreservation of buffalo sperm in Bangladesh. This study aimed to evaluate the suitability of different semen extenders for improving the post-thaw quality of Bangladeshi buffalo sperm under a manual cryopreservation protocol. The manual cryopreservation protocol was compared and optimized with a commercial biofreezer protocol. Then, the efficacy of different diluters was evaluated using the optimized manual cryopreservation protocol. Meanwhile, the post-thaw sperm quality in terms of motility and morphology was evaluated by CASA during the optimization process. During manual cryopreservation, the first cooling from 37 °C to 5 °C was done in a BLRI-developed equilibration chamber and the second cooling from 5 °C to -120 °C in a Styrofoam box using liquid nitrogen vapor from different distances (0.5, 1.5, 1.6, 2 and 3 inches). Simultaneously, another batch of sperm was cryopreserved using a programmable freezer. The highest number of motile sperms (62.67±1.12; P&lt;0.01) and progressive motility (38.97±1.10; P&lt; 0.001) was observed at 1.6 inches above liquid nitrogen, which are similar to the results obtained from automated biofreezer protocol (65.94±4.65 and 45.54 ± 3.64, respectively). To evaluate the semen extenders’ efficacy, one locally developed Tris-fructose-egg yolk-based diluter and two commercial diluters (Andromed, Triladyl) were used in the freezing of buffalo sperm. The highest recovery and conception rates were observed in sperm diluted with TFE (82.4% and 80%, respectively). Therefore, it is clear that this manual cryopreservation protocol and the TFE diluter could be a suitable and inexpensive alternative for Bangladeshi buffalo sperm considering post-thaw sperm quality and fertility.

Cpne1 deficiency preserves sperm motility under Ca2+ channel blockade.

Calcium ions (Ca2+) play crucial roles in sperm motility and fertilization. The copine (CPNE) family comprises several Ca2+-dependent phospholipid-binding proteins. Of these, CPNE1 is extensively expressed in mammalian tissues; however, its precise role in testicular development and spermatogenesis is yet to be fully characterized. In this study, we used proteomics to analyze testicular biopsies and found that levels of CPNE1 were significantly reduced in patients with non-obstructive azoospermia (defective spermatogenesis) compared to those in patients with obstructive azoospermia (physiological spermatogenesis). In mice, CPNE1 is expressed at various stages of germ cell development and is associated with the Golgi apparatus. Ultimately, CPNE1 is expressed in the flagella of mature sperms. To further examine the role of CPNE1, we developed a Cpne1 knockout mouse model. Analysis showed that the loss of Cpne1 did not impair testicular development, spermatogenesis, or sperm morphology and motility in physiological conditions. When treated with gadolinium (III) chloride or 2-aminoethoxydiphenyl borate, known inhibitors of store-operated Ca2+ entry, Ca2+ signals and sperm motility were significantly compromised in wild-type mice; however, both mechanisms were conserved in KO mice. These results suggested that CPNE1 is dispensable for testicular development, spermatogenesis or sperm motility in physiological conditions. In addition, CPNE1 may represent a target of Ca2+ channel inhibitors and may therefore be implicated in the regulation of Ca2+ signaling and sperm motility.

Effects of liquid storage of buck semen at refrigeration temperatures on sperm viability and fertility to develop ready to use goat semen diluent

It is generally acknowledged that lower pH throughout liquid storage can decrease sperm motility and energy production, which may be advantageous for maintaining sperm viability during cryopreservation. Hence, the purpose of this study was to see how pH affected sperm viability, motility, membrane integrity, and oxidative changes after 5 days of storage in chemically specified extenders at refrigerator temperatures to formulate a ready-to-use goat semen diluent. Ejaculates were extended with tris-citric acid-fructose diluent with 20% or without egg yolk and stored at 5°C for 5 days. Sperm motility, live count, acrosome integrity, hypo-osmotic swelling positive spermatozoa, pH, and malondialdehyde all substantially differed significantly (P &lt; 0.05) during different days of liquid storage. The pH of diluted semen decreased considerably (P &lt; 0.05) from its original pH of 6.46 on 0 day to pH 6.06 on the 5th day of liquid storage at refrigeration temperature. Diluted semen at refrigeration temperature having 20% egg yolk had significantly higher (P &lt; 0.05) seminal characteristics compared to liquid semen without egg yolk. The liquid storage of semen quality became worse day by day due to a drop in pH and egg yolks acted as a cryoprotectant against the harmful effects of chilling. Hence, 20% egg yolk may be used to enhance the keeping quality of diluted buck semen at refrigeration temperature for up to 5 days. Based on this result, we formulated a ready to use goat semen diluent for liquid semen storage at room temperature and refrigeration temperature.

Role of sperm morphological parameters in the selection of fertilization methods.

To investigate the role of in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) in cases with isolated teratozoospermia, and to provide reference for infertile patients to choose a reasonable insemination program. From January 2018 to December 2022, 433 couples received assisted reproductive assistance in the Department of Reproductive Medicine of our hospital. Among them, 81 patients presented isolated teratozoospermia (other sperm parameters were normal). These underwent IVF/ICSI, and differences in embryological data were compared between groups. 60 patients with moderate teratozoospermia went IVF, 11 needed rescue ICSI (R-ICSI, 18.33%), and 49 developed up to the blastocyst stage. 14 patients with severe teratozoospermia went IVF, 2 needed R-ICSI (14.29%), and 12 developed up to the blastocyst stage. There were no statistically significant differences in the proportion of R-ICSI between the two groups. Embryological outcomes were not significantly different between groups. The present results suggest that patients with moderate and severe teratozoospermia can be effectively treated with IVF, being R-ICSI performed when necessary.

Clinical Benefits of Decreased Photo-Oxidative Stress on Human Embryo Development

Objective: Early embryonic development is characterized by rapid cell division and gene activation, making the embryo extremely sensitive to environmental influences. Light exposure can affect embryonic development through a direct toxic effect on the embryo via the generation of reactive oxygen species. In a previous study, we demonstrated the positive effect of improved light-protected embryo culture conditions implemented in our laboratory. This study aimed to investigate the changes in human embryo development under light protection during the conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). Materials and Methods: We tested the potential beneficial effect of light filters to reduce the risk of toxic effects of light. IVF outcomes were compared between two experimental conditions, light protection with red light filters versus no light protection as a control. Results: Blastocyst development rate in IVF was significantly higher in the light-protected group than in the group treated under conventional conditions (46.6 vs. 26.7%). In the case of ICSI, we obtained a similar result (44.5 vs. 31.6%). The rate of cryopreservation with at least one embryo was higher in the light-protected phase (32.8%) than in the conventionally manipulated phase (26.8%). The abortion rate was also significantly lower during the light-protected period in IVF, resulting in a higher live birth rate. Conclusions: The implementation of light protection to reduce the embryotoxic wavelengths of light in IVF centers may improve the blastocyst development rate and embryo quality while maintaining embryo safety.