Sperm Loss Research Articles

Revisiting cadmium-induced toxicity in the male reproductive system: an update.

Heavy metals like cadmium (Cd) are one of the main environmental pollutants, with no biological role in the human body. Cd has been well-documented to have disastrous effects on both plants and animals. It is known to accumulate in kidneys, lungs, liver, and testes and is thought to affect these organs' function over time, which is linked to a very long biological half-life and a very poor rate of elimination. According to recent researches, the testes are extremely vulnerable to cadmium. The disruption of the blood-testis barrier, seminiferous tubules, Sertoli cells, and Leydig cells caused by cadmium leads to the loss of sperm through various mechanisms, such as oxidative stress, spermatogenic cell death, testicular swelling, dysfunction in androgen-producing cells, interference with gene regulation, disruption of ionic homeostasis, and damage to the vascular endothelium. Additionally, through epigenetic control, cadmium disrupts the function of germ cells and somatic cells, resulting in infertile or subfertile males. A full grasp of the mechanisms underlying testicular toxicity caused by Cd is very important to develop suitable strategies to ameliorate male fertility. Therefore, this review article outlines cadmium's impact on growth and functions of the testicles, reviews therapeutic approaches and protective mechanisms, considers recent research findings, and identifies future research directions.

Genital courtship and female-active roles in mating: sexual selection by mate choice in Caenorhabditis elegans.

A new bridge between studies of sexual selection and the massive literature on Caenorhabditis elegans behaviourand nervous system properties promise to provide important new insights into both fields. This paper shows that mate choice likely occurs in hermaphrodite C. elegans on the basis of stimulation from the male genital spicules, making it possible to apply the toolkit of extensive background knowledge of C. elegans and powerful modern techniques to test in unprecedented detail the leading hypotheses regarding one of the most sweeping trends in all of animal evolution, the especially rapid divergence of genital morphology. The recognition that sexual selection by mate choice may also occur in other contexts in C. elegans suggests additional payoffs from exploring previously unrecognized possibilities that female-active hermaphrodite reproductive behaviours are triggered by male stimulation. These facultative behaviours include attracting males, fleeing from or otherwise resisting males, opening the vulva to allow intromission, guiding sperm migration, avoiding rapid oviposition following copulation that results in sperm loss, expelling recently received sperm, and increasing feeding rates following copulation.

Targeting mosquito X-chromosomes reveals complex transmission dynamics of sex ratio distorting gene drives

Engineered sex ratio distorters (SRDs) have been proposed as a powerful component of genetic control strategies designed to suppress harmful insect pests. Two types of CRISPR-based SRD mechanisms have been proposed: X-shredding, which eliminates X-bearing sperm, and X-poisoning, which eliminates females inheriting disrupted X-chromosomes. These differences can have a profound impact on the population dynamics of SRDs when linked to the Y-chromosome: an X-shredder is invasive, constituting a classical meiotic Y-drive, whereas X-poisoning is self-limiting, unable to invade but also insulated from selection. Here, we establish X-poisoning strains in the malaria vector Anopheles gambiae targeting three X-linked genes during spermatogenesis, resulting in male bias. We find that sex distortion is primarily driven by a loss of X-bearing sperm, with limited evidence for postzygotic lethality of female progeny. By leveraging a Drosophila melanogaster model, we show unambiguously that engineered SRD traits can operate differently in these two insects. Unlike X-shredding, X-poisoning could theoretically operate at early stages of spermatogenesis. We therefore explore premeiotic Cas9 expression to target the mosquito X-chromosome. We find that, by pre-empting the onset of meiotic sex chromosome inactivation, this approach may enable the development of Y-linked SRDs if mutagenesis of spermatogenesis-essential genes is functionally balanced.

Pre-freezing selection of Holstein bull semen with the BoviPure colloid as double- or single-layer centrifugation improves the post-thawing quality

Artificial insemination (AI) is critical for breeding in the dairy industry. High-merit bulls can present low freezability, hampering genetic dissemination. Thawed semen can be improved using density gradient centrifugation (DGC) with colloids, but little information deals with the pre-freezing application. Thus, the BoviPure colloid (optimized for bull spermatozoa) was tested for pre-freezing application as the usual double-layer (DLC) versus single-layer (SLC, quick and economical). Semen from twelve Holstein-Friesian bulls was extended with OPTIXcell extender, frozen (Control), or processed by SLC or DLC and frozen. Sperm were assessed pre-freezing for motility and viability and post-thawing (directly and after 4 h 38 °C) for apoptosis, capacitation status, acrosomal damage, mitochondrial activity, cytoplasmic and mitochondrial reactive oxygen species (ROS), and chromatin status (SCSA for DNA fragmentation and chromatin compaction and monobromobimane, mBBr, for disulfide bridges evaluation). The DGC improved parameters post-thawing (e.g., 57.5%±10.1 motility vs. control 53.3% ± 11.2) at the cost of sperm loss (sperm recovery of DGC 14.4% ± 2.5 and SLC 17.4% ± 2.5). DNA fragmentation (%DFI) decreased (0.21% ± 0.53 vs. control 1.30% ± 0.10), and SLC reduced chromatin compaction. A clustering procedure separated lesser (LF) and greater freezability (GF) bulls. LF samples were especially benefited by DGC, with SLC providing better post-thawing results for this group. In conclusion, pre-freezing DGC improved sperm parameters post-thawing, potentially improving the cryopreservation of low-freezability semen from high-merit bulls. SLC, quicker and economical, would be preferable since it showed similar or higher performance than DLC.

Decreased AKAP4/PKA signaling pathway in high DFI sperm affects sperm capacitation.

The sperm DNA fragmentation index (DFI) is a metric used to assess DNA fragmentation within sperm. During in vitro fertilization-embryo transfer (IVF-ET), high sperm DFI can lead to a low fertilization rate, poor embryo development, early miscarriage, etc. A kinase anchoring protein (AKAP) is a scaffold protein that can bind protein kinase A (PKA) to subcellular sites of specific substrates and protects the biophosphorylation reaction. Sperm protein antigen 17 (SPA17) can also bind to AKAP. This study intends to explore the reason for the decreased fertilization rate observed in high sperm DFI (H-DFI) patients during IVF-ET. In addition, the study investigates the expression of AKAP, protein kinase A regulatory subunit (PKARII), and SPA17 between H-DFI and low sperm DFI (L-DFI) patients. SPA17 at the transcriptional level is abnormal, the translational level increases in H-DFI patients, and the expression of AKAP4/PKARII protein decreases. H 2 O 2 has been used to simulate oxidative stress damage to spermatozoa during the formation of sperm DFI. It indicates that H 2 O 2 increases the expression of sperm SPA17 protein and suppresses AKAP4/PKARII protein expression. These processes inhibit sperm capacitation and reduce acrosomal reactions. Embryo culture data and IVF outcomes have been documented. The H-DFI group has a lower fertilization rate. Therefore, the results indicate that the possible causes for the decreased fertilization rate in the H-DFI patients have included loss of sperm AKAP4/PKARII proteins, blocked sperm capacitation, and reduced occurrence of acrosome reaction.

Effects of grape seed proanthocyanidin extract on side effects of high-dose methylprednisolone administration in male rats.

In this study, we investigated the effects of grape seed proanthocyanidin extract (GSPE) against the side effects of high-dose administration of methylprednisolone (MP) in male rats. A total of 32 adult Wistar male albino rats were divided into four groups: (1) control (CON), received standard food only; (2) MP, received standard food + intraperitoneal injection of 60mg/kg MP on day 7; (3) GSPE, received standard food + 200mg/kg/day GSPE; and (4) MP + GSPE, received standard food + 200mg/kg/day of GSPE + intraperitoneal injection of 60mg/kg MP on day 7. All animals in the GSPE and GSPE + MP groups were treated once a day by oral gavage for 14 consecutive days. The feed intake of rats in the MP and MP + GSPE groups decreased significantly by 24.14% and 13.52%, respectively (p < 0.05). Administration of MP resulted in significant increases in serum concentrations of blood urea nitrogen (p < 0.001), glucose (p < 0.01), alkaline phosphatase, and adrenocorticotropic hormone (p < 0.05). High-dose MP administration significantly reduced catalase (p < 0.001) and glutathione peroxidase (p < 0.05) concentrations in the liver and kidney tissues of rats, while glutathione concentrations were only reduced in liver tissue (p < 0.05). The expression levels of Bcl-2 and TNF-α in liver, kidney, and testicular tissue were significantly increased, while the expression levels of caspase-3 were reduced (p < 0.001). Furthermore, sperm concentration was significantly affected by GSPE in rats induced by high-dose MP, and sperm loss was significantly reduced in MP + GSPE (p < 0.05). These findings suggest that GSPE could be useful as a supplement to alleviate MP-induced toxicity in rats.

Defining an Optimal Range of Centrifugation Parameters for Canine Semen Processing.

Our objective was to determine a clinically relevant range of centrifugation parameters for processing canine semen. We hypothesized that higher gravitational (g) force and longer time of centrifugation would result in improved spermatozoa recovery rate (RR) but poorer semen quality. Cooled storage under standard shipping conditions was used as a stressor to evaluate long-term treatment effects. Individual ejaculates collected from 14 healthy dogs were split into six treatment groups (400 g, 720 g, and 900 g for 5 or 10 min). Sperm RR (%) was calculated post-centrifugation, and plasma membrane integrity (%, Nucleocounter® SP-100™), total and progressive motility (%, subjective and computer-assisted sperm analysis), and morphology (%, eosin-nigrosin staining) were assessed on initial raw semen (T0), post-centrifugation (T1), and 24 h (T2) and 48 h (T3) after cooling. Sperm losses were minimal, and RRs were similar across treatment groups (median >98%, p ≥ 0.062). Spermatozoa membrane integrity was not different between centrifugation groups at any time point (p ≥ 0.38) but declined significantly during cooling (T1 vs. T2/T3, p ≤ 0.001). Similarly, total and progressive motility did not differ across treatments but declined in all groups from T1 to T3 (p ≤ 0.02). In conclusion, our study showed that centrifugation within a range of 400 g-900 g for 5-10 min is appropriate for processing canine semen.

Panax ginseng C.A. meyer alleviates benign prostatic hyperplasia while preventing finasteride-induced side effects.

Panax ginseng C.A. Meyer, a widely used traditional medicine in East Asia, shows many beneficial effects on immune function, male erectile dysfunction, cancer, excessive oxidants, and aging issues. However, its effect on benign prostatic hyperplasia (BPH) and its potential in the treatment of side effects related to finasteride (Fi), an FDA-approved drug for BPH, are less known. This study aimed to verify the therapeutic effects of a water extract of P. ginseng (PGWE) on BPH in testosterone propionate (TP)-induced BPH rats and TP-treated RWPE-1 human epithelial cells, and the inhibitory potential on the Fi-induced side effects is also explored. In the TP-induced BPH rat model, PGWE alleviated the pathological markers of BPH such as weight and epithelial thickness of the prostate, and the serum level of dihydrotestosterone. PGWE downregulated androgen-related BPH factors such as 5α-reductase 2 and androgen receptor. PGWE also showed prostatic cell apoptosis accompanied by increased expression of Bax and decreased expression of Bcl-xL and cleaved-caspase 3, respectively, in addition to increasing mitochondrial dynamics in both in vivo and in vitro BPH models. Notably, reduced sperm count, one of the serious side effects of Fi, in the epididymis of BPH rats was recovered with PGWE treatment, suggesting less toxicity to sperm development by PGWE. PGWE also protected against Fi-induced sperm loss when PGWE was administered in combination with Fi without compromising the therapeutic effects of Fi on BPH. Based on these findings, we propose that PGWE could be an alternative therapeutic agent for BPH.

Lipid alterations by oxidative stress increase detached acrosomes after cryopreservation of semen in Holstein bulls

The nearly exclusive use of cryopreserved semen in cattle breeding enables long shipping distances, higher storage times, quarantine to avoid germ transmission and easy dispersal of high genetic value bulls. Spermatozoa from bulls are well freezable and improvement of cryopreservation protocols over decades has led to high semen quality. However, there is still some loss of spermatozoa in each semen dose due to detached acrosomes after thawing. There are even individual bulls with extremely high numbers of detached acrosomes after cryopreservation, called “bad freezers”. This study screened 1092 ejaculates from 59 Holstein bulls for the difference in detached acrosomes before and after cryopreservation (ΔAC). The individual bull influenced ΔAC (P < 0.001) and allowed selection for individuals with repeatedly low ΔAC (good freezers) or high ΔAC (bad freezers). Good freezers were superior to bad freezers in a thermo-resistance test (78.2% vs. 33.6% total motility, respectively, P = 0.047) and had higher non-return rates (NRR: 46.8% vs. 40.8%, respectively, P = 0.016). Since oxidative stress is one possible explanation for premature acrosome reaction, the radical reduction capacity of the seminal fluid was measured, finding that this parameter was reduced in bad freezer bulls during cryopreservation (P = 0.043). Analysis of lipid species in sperm cells by matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) showed a reduction of ether lipids and plasmalogens as well as an increase in formyl-lysophosphatidylcholines only within the bad freezers during cryopreservation (P = 0.043). In conclusion these findings show, that lipid alteration caused by oxidative stress is one essential reason for highly augmented acrosome reacted spermatozoa in bad freezer bulls. Therefore, increased use of antioxidants in the extender could be a possible starting point for developing individualized extenders for bad freezer bulls of high genetic value, in order to raise sperm quality after cryopreservation even in those bulls.

Mechanisms of Cadmium-Induced Testicular Injury: A Risk to Male Fertility.

Cadmium is a heavy toxic metal with unknown biological functions in the human body. Over time, cadmium accretion in the different visceral organs (liver, lungs, kidney, and testis) is said to impair the function of these organs, which is associated with a relatively long biological half-life and a very low rate of excretion. Recently studies have revealed that the testes are highly sensitive to cadmium. In this review, we discussed the adverse effect of cadmium on the development and biological functions of the testis. The Sertoli cells (SCs), seminiferous tubules, and Blood Testis Barrier are severely structurally damaged by cadmium, which results in sperm loss. The development and function of Leydig cells are hindered by cadmium, which also induces Leydig cell tumors. The testis's vascular system is severely disturbed by cadmium. Cadmium also perturbs the function of somatic cells and germ cells through epigenetic regulation, giving rise to infertile or sub-fertile males. In addition, we also summarized the other findings related to cadmium-induced oxidative toxicity, apoptotic toxicity, and autophagic toxicity, along with their possible mechanisms in the testicular tissue of different animal species. Consequently, cadmium represents a high-risk factor for male fertility.

An optimized centrifugation protocol for ram sperm ensuring high sample yield, quality and fertility

The optimization and implementation of artificial insemination (AI) in sheep is necessary to increase the livestock productivity through enhanced control of reproductive function. Sperm centrifugation is a common procedure in the ejaculate handling in AI and other assisted reproductive technologies (ART), as part of new methods of sperm analysis, selection or preservation. However, our research group previously established that this simple procedure might cause a large sperm loss and induce deleterious effects on the sperm function of the ovine species when high centrifugation forces are employed. To our knowledge, there are no studies on combined effect of extender and different centrifugal forces on ram sperm yield and quality. Furthermore, evidence of in vivo fertility rate using sperm obtained with various centrifugation forces is also lacking in this species. Thus, the objective of this work was to define the ideal conditions for ram semen centrifugation that will achieve the best quantity and quality sample to ensure unaffected fertilization ability of centrifuged ram sperm. The Experiment 1 evaluated the effect of the centrifugation procedure of two extenders (INRA 96 and Tyrode's) and two cooling protocols (Rapid and Slow Refrigeration −35 °C to 15 °C–) on sperm recovery rate and quality (motility and kinetic parameters, viability, apoptosis and mitochondrial activity). INRA 96 combined with Slow Refrigeration and Tyrode's at room temperature registered the highest sperm recovery and quality values (P ≤ 0.05). In Experiment 2, the influence of three centrifugal forces (600, 1200 and 6000×g for 10 min) was assessed immediately after centrifugation on the technical performance and sperm functionality in diluted samples with INRA 96 and Tyrode's at the conditions set out in Experiment 1. The lowest pellet weight (P ≤ 0.05) without harmful effect on sperm physiological status (P > 0.05) was achieved at 1200×g, since 6000×g induced sperm motility damage (P ≤ 0.05) with both extenders. Finally, to ensure the total safety of the centrifugation protocol, Experiment 3 tested in a combined in vitro and in vivo test the effect of these three centrifugal forces on ram sperm quality after dilution (INRA 96) and liquid storage (6–8 h at 15 °C). The damage produced by 6000×g on sperm motility (P ≤ 0.05) was maintained over time, coinciding with a lower fertility (P ≤ 0.05). In conclusion, ram sperm can be centrifuged in INRA 96 extender up to 1200×g for 10 min at 15 °C as secure values with high recovery rates and without detrimental effects on sperm quality and fertility.

Histomorphometric and Histopathological Alterations of Rat Testis Following Exposure to Hydrogen ‎Peroxide: Protective Role of Resveratrol Supplement

&#x0D; &#x0D; &#x0D; &#x0D; Resveratrol (RS) is widely used in medical fields as an antioxidant. Current research investigated ‎the protective role of RS supplement on histomorphometric and histopathological alterations in ‎testes were caused by hydrogen peroxide (H2O2) of rats as an animal model. Thirty-two adult ‎rats were utilized in this study, divided randomly into 4 equal groups as follows. The group C ‎was given tap water only and served as control, the 2nd group (G1) was given 0.5% of H2O2 of ‎tap water, the 3rd group (G2) was given tap water containing 0.5% of H2O2 and intubated RS ‎supplement at 87 mg/kg BW, and 4th group intubated RS supplement 87 mg/kg BW. after 56 ‎days of treatment, rats were euthanized, dissected then, specimens of testes tissue were collected ‎for histomorphometric and histopathological evaluation. Our results showed that administration ‎of H2O2 caused a significant histomorphometric with histopathological changes in the form of a ‎thickness of fibrous tunica albuginea, disarrangement of germ cells, necrosis of spermatogonia, ‎edema, and loss of sperms as compared to other groups. Meanwhile, these histological alterations ‎were partially attenuated in the G2 group that intubated resveratrol. Thus, the current study ‎concluded that resveratrol may have therapeutic value in the treatment of induced testicular injury ‎by H2O2 due to its antioxidant activity and attenuation of harmful effects of oxidative ‎stress through a mechanism that should be elucidated in future studies.&#x0D; &#x0D; &#x0D; &#x0D;

Minimum sperm dose for optimal fertility after artificial insemination in ostriches

Artificial insemination (AI) in ostriches may present potential solutions to high proportions of infertile eggs commonly recorded on commercial farms and assist in reducing the number of males for breeding purposes thereby leading to a more economical and efficient farming system. Although non-invasive methods to collect semen and to artificially inseminate female ostriches have been developed, the insemination dose for maximum fertility of eggs remains unknown. This study was thus conducted to determine the minimum sperm dose that would promote fertility of eggs following AI in female ostriches. A total of 22 South African black ostriches (7 males and 15 females) aged between 2 and 9 years old were used. Semen samples were collected using the dummy female method and diluted 1:4 (semen: diluent) with an ostrich specific semen diluent. Females were inseminated with various sperm doses of diluted semen from the same male three times a week, every second day resulting in a total sperm dose of A: <2.5 × 109, B: 2.5–5 × 109, C: 5–7.5 × 109 and D: 7.5–9.6 × 109 sperm/week. Eggs produced after insemination were opened to determine the fertilization status of the germinal disc (GD) with an unaided eye. The fertile period was then calculated as the number of days fertilized eggs were laid after the last AI. Furthermore, the number of sperm trapped in the outer perivitelline membrane (spermOPVL) above the GD region was counted under fluorescent light, following staining with 4′,6-diamidino-2-phenylindole to determine the rate of sperm loss and the number of days up to when the last egg containing sperm was laid. On average, a mean (±sd) of 35.34 ± 25.72% eggs produced after AI were fertilized. Fertility was lower (mean ± se) when sperm dose A was used (6.71 ± 9.40%), as compared to sperm dose B (46.01 ± 6.71%), C (37.34 ± 6.60%) and D (37.75 ± 8.36%) (P < 0.05). No significant difference was recorded in the latter three doses (P > 0.05). Furthermore, the length of the fertile period and the rate of sperm loss did not differ significantly between sperm doses (P > 0.05). Hence, a sperm dose of between 2.5 and 5 × 109 sperm/week is recommended to optimize fertility after AI in ostriches, as increasing the sperm dose would not benefit fertility. Further studies are, however, needed to determine the frequency of insemination that would maintain fertility throughout the breeding season as well as hatchability of eggs laid after AI.

Advantages of filtration method for sperm-DNA genotyping in sexual assault cases.

Differential extraction (DE) is a conventional method to isolate sperms from forensic semen samples (e.g. vaginal swab containing semen) for sperm-DNA genotyping. Subsequent to selective digestion of somatic cells in a mixture sample, sperms are collected and purified as a pellet by repetitive centrifugation based on the specific gravity of sperm heads. However, the centrifugation operation requires a technical proficiency and an extensive time to prevent a loss of sperms from the pellet as much as possible. Therefore, we devised a "filtration method (FM)", in which a vacuum filtration operation based on the size of sperm heads is adapted, instead of DE, for isolation of sperms without any loss in principle. Sperms are collected and purified on a polycarbonate membrane filter. In this study, we have compared results of forensic assays by DE and FM for sperm-DNA genotyping from forensic semen samples. Consequently, FM had advantages of easy operation, timesaving, and high yield of sperms from semen samples compared with DE, although FM had a comparable ability to DE for a purification of sperms from mixture samples. Thus, we present that FM could simply lead to success of sperm-DNA genotyping and has a possibility to supersede DE as a gold-standard method.

Tetrapeptides Modelled to the Androgen Binding Site of ZIP9 Stimulate Expression of Tight Junction Proteins and Tight Junction Formation in Sertoli Cells.

Simple SummaryAndrogens such as testosterone act either through the nuclear androgen receptor or through the plasma membrane-bound androgen receptor ZIP9. Androgens are crucial for male fertility. In the testis they stimulate the formation of tight junctions between Sertoli cells, thereby creating an immune-privileged environment for the development of fertile sperm known as the blood–testis barrier. Androgens are also absolutely required for spermatogenesis. Nevertheless, when low testosterone is supplemented by exogenous testosterone (e.g., in testosterone replacement therapy), it triggers a loss of sperm, and thus and infertility, by a negative feed-back mechanism based on the hypothalamic-pituitary-gonadal axis. In order to investigate this discrepancy, the effects of small peptides that fit within the androgen binding site of ZIP9 were tested for their effects on the expression of proteins involved in the formation of tight junctions. Our results provide evidence that all three peptides tested act through ZIP9 as androgens and promote tight junction protein expression and tight junction formation. They can therefore act as surrogates of testosterone for the formation and maintenance of the blood–testis barrier.Androgens stimulate the expression of tight junction (TJ) proteins and the formation of the blood–testis barrier (BTB). Interactions of testosterone with the zinc transporter ZIP9 stimulate the expression of TJ-forming proteins and promote TJ formation in Sertoli cells. In order to investigate androgenic effects mediated by ZIP9 but not by the nuclear androgen receptor (AR), the effects of three tetrapeptides fitting the androgen binding site of ZIP9 were compared with those induced by testosterone in a Sertoli cell line expressing ZIP9 but not the AR. Three tetrapeptides and testosterone displaced testosterone-BSA-FITC from the surface of 93RS2 cells and stimulated the non-classical testosterone signaling pathway that includes the activation of Erk1/2 kinases and transcription factors CREB and ATF-1. The expression of the TJ-associated proteins ZO-1 and claudin-5 was triggered as was the re-distribution of claudin-1 from the cytosol to the membrane and nucleus. Furthermore, TJ formation was stimulated, indicated by increased transepithelial electrical resistance. Silencing ZIP9 expression by siRNA prevented all of these responses. These results are consistent with an alternative pathway for testosterone action at the BTB that does not involve the nuclear AR and highlight the significant role of ZIP9 as a cell-surface androgen receptor that stimulates TJ formation.

Rapid cryopreservation of small quantities of human spermatozoa by a self-prepared cryoprotectant without animal component.

Cryopreservation of small quantities of human spermatozoa whilst maintaining adequate post‐thawing motility has been found an essential challenge for male fertility preservation. Therefore, the study used an effective, and convenient rapid‐freezing method to freeze small amounts of human spermatozoa by adding self‐prepared cryoprotectant (SPC) without animal component. In the feasibility experiment, no significant differences in progressive motility, normal sperm morphology, vitality or DNA fragmentation index between the conventional slow freezing and rapid freezing were realised. The present study prospectively analysed the effects of sperm freezing and resuscitation in 175 patients with severe oligozoospermia (sperm concentration <1 × 106/ml). We observed the 120 severe oligozoospermia specimens had a mean recovery rate of 60.19% ± 10.43% and a mean cryosurvival rate of 68.0% ± 9.17%. In addition, 55 cryptozoospermia specimens were analysed. The small‐volume cryopreservation showed advantages. The total sperm recovery, motility recovery and sperm loss rates were 98.48%, 50.17% and 1.52% respectively. In short, the SPC is safe and effective, and can be used to rapidly freeze severe oligozoospermia specimens. That is useful for successful sperm freezing whilst avoiding the risk of azoospermia in the later stages and promoting comprehensive fertility preservation.

EBV induced loss of sperm quality.

To analyze the presence of Epstein-Barr-virus (EBV) in sperm samples from patients diagnosed with some impairment of the fertility parameters evaluated using seminogram and to observe if there is any difference with the normozoospermic samples. We hypothesize that an EBV infection is responsible for the upregulation of the miRNA 199-3p, which binds to the 3'UTR of endothelin-1 (ET-1). ET-1 is a key factor to produce Vimentin (Vim3), and therefore, it influences the expression of Vim3. Since Vim3 is predominantly detectable in sperms without any structural defects, the newly identified regulation mechanism can be responsible for the loss of sperm quality. This study was performed from January 2017 to December 2020 and included 27 patients who provided ejaculated samples obtained by masturbation. Ejaculates were evaluated according to the Word Health Organization's criteria. Posteriorly, the samples were sorted according to the seminogram diagnosis and further analyzed using different enzyme-linked absorbed immune assays to determine the level or concentration of Epstein-Barr nuclear antigen (EBNA), ET-1, and Vim3. All sperm samples with the impairment of fertility parameters contained the EBNA and presented a downregulation of ET-1 and Vim3. In addition, sperms located in the swim ups are also partially positive for the EBV virus in different clinical aspects. Based on the regulation mechanism here presented, it seems that the EBV induces changes at the miRNA level, which are responsible for the decreasing of sperm quality.

Heterotrimeric Kinesin II is required for flagellar assembly and elongation of nuclear morphology during spermiogenesis in Schmidtea mediterranea

Development of sperm requires microtubule-based movements that drive assembly of a compact head and flagellated tails. Much is known about how flagella are built given their shared molecular core with motile cilia, but less is known about the mechanisms that shape the sperm head. The Kinesin Superfamily Protein 3A (KIF3A) pairs off with a second motor protein (KIF3B) and the Kinesin Associated Protein 3 (KAP3) to form Heterotrimeric Kinesin II. This complex drives intraflagellar transport (IFT) along microtubules during ciliary assembly. We show that KIF3A and KAP3 orthologs in Schmidtea mediterranea are required for axonemal assembly and nuclear elongation during spermiogenesis. Expression of Smed-KAP3 is enriched during planarian spermatogenesis with transcript abundance peaking in spermatocyte and spermatid cells. Disruption of Smed-kif3A or Smed-KAP3 expression by RNA-interference results in loss of spermatozoa and accumulation of unelongated spermatids. Confocal microscopy of planarian testis lobes stained with alpha-tubulin antibodies revealed that spermatids with disrupted Kinesin II function fail to assemble flagella, and visualization with 4’,6-diamidino-2-phenylindole (DAPI) revealed reduced nuclear elongation. Disruption of Smed-kif3A or Smed-KAP3 expression also resulted in edema, reduced locomotion, and loss of epidermal cilia, which corroborates with somatic phenotypes previously reported for Smed-kif3B. These findings demonstrate that heterotrimeric Kinesin II drives assembly of cilia and flagella, as well as rearrangements of nuclear morphology in developing sperm. Prolonged activity of heterotrimeric Kinesin II in manchette-like structures with extended presence during spermiogenesis is hypothesized to result in the exaggerated nuclear elongation observed in sperm of turbellarians and other lophotrochozoans.

NGF Rescues Spermatogenesis in Azoospermic Mice.

Nerve growth factor (NGF) plays an important role in regulating the hypothalamus-pituitary-gonadal (HPG) axis. However, the effects of NGF on spermatogenesis remain unclear. This study aimed to assess the potential application of NGF with nasal delivery on spermatogenesis in azoospermic mice. We established a model with azoospermia induced by a single intraperitoneal (i.p.) injection of busulfan. NGF pre-encapsulated with liposomes (25, 50, and 100 μg/kg) was delivered via internasal administration. Three weeks after busulfan injection, NGF treatments were performed twice a week for 8 weeks; the change of sperm quality, testis and epididymis histopathology, and androgenic hormone were analyzed to evaluate sperm regeneration. Furthermore, 30 mg/kg busulfan injection caused severe testicular atrophy of the seminiferous tubules, characterized by a loss of spermatogenic elements and sperms. NGF with nasal administration could significantly upregulate the markers expressing meiotic spermatogonia (Stra8) and spermatocytes (SYCP3), restore spermatogenesis, and improve sperm quality in busulfan-treated mice by increasing the secretion of sexual hormones. The convenient and noninvasive nasal delivery of NGF may be a new potential therapy for spermatogenesis via activating the HPG axis and elevating androgenic hormones. This study opened a new horizon for NGF application in reproductive endocrine.

The Impact of Gemcitabine-induced Reproductive Toxicity on The Sperm count and Morphology of Albino Rats

Cancer treatments can affect sperm production and a significant percentage of cancer patients may develop permanent azoospermia or severe oligozoospermia after chemotherapy. To investigate the influence of Gemcitabine toxicity on the reproductive system of albino male rats (sperm count and morphology). An experimental animal study conducted in the zoology department, College of Science, King Saud University during the period from June to October 2014 using albino rats (Rattusnorvegicus) (Wistar strain). Males were divided into four different groups (control" 0 mg/kg",7 mg/kg,14 mg/kg, and 21 mg/kg). The reproductive organs, testicles and epididymis decreased in weight and atrophied in most of the animals treated with the drug in various doses. The mean absolute and relative epididymal weights were also significantly decreased. In the drug-effects recovery group, neither the testicles nor the epididymis in the animals treated with the three doses recovered fully normal weight. The testis's efficiency in producing sperm was significantly decreased at all doses. In the recovery group, the testis regained its efficiency, as no significant difference was recorded between the drug-treated groups and the control group. The drug caused complete loss of sperm, in a rat treated with the big dose. Gemcitabine caused a significant increase in the percentage of deformed sperms in all treated animals. Gemcitabine drug has high toxicity on the reproductive system of rats with a dose tenth of human dose, with a massive decrease in the count and quantity of sperm, which means that this drug can have more toxicity effects on human.