Sperm Function Research Articles

Evaluation of the Detrimental Impact of Low-Intensity Laser Radiation on the Characteristics of Sperm Movement, Motion, and DNA Damage

It is generally accepted that low-level laser therapy promotes cellular energy production and increases the synthesis of ATP by triggering the mitochondrial electron transport chain's photosensitive cytochrome c oxidase complex. Nevertheless, this investigation aimed to examine the possible adverse impact on sperm function and DNA integrity of low-intensity laser radiation with a wavelength of 980 nm. Following standard analysis, forty semen samples were collected and categorized as either asthenospermic, oligospermic, or normospermic. The remaining semen was subjected to standard semen analysis, and the aliquots were divided into treatment and control groups. A continuous-wave 980 nm laser with an output power of 100 mW and an energy density of 10 J/cm2 was applied to the treated samples for 30 s. Divided samples underwent incubation at 37oC, and semen analysis was used to assess aliquots at 30 s and 2 h intervals. Following the incubation time, each sample was frozen in 250 mL at -80oC until DNA fragmentation was examined using flow cytometry. Thirty minutes after treatment, there was a noticeable increase in sperm movement; the rise was largest in oligospermic and asthenospermic samples (88%) while samples of normal sperm displayed the least increment (7.5%). There was no discernible increase in DNA damage in the treatment samples compared to the control samples. The treatment samples had 21.8% and the control samples had 25.3% of the DNA fragmentation index. The dynamics of sperm motion were noticeably altered. Short-term exposure to low-level laser radiation appeared to improve the motion and movement of treated sperm, and after 2 h, there was no increase in DNA damage. Asthenospermia and mitochondrial dysfunction may be related in some circumstances. To fully understand the ramifications of these results for possible medicinal applications, more research is needed. Our research's conclusions suggest that low-level laser stimulation is a useful and safe technique for identifying viable immotile spermatozoa in a particular sample. It may also cause these spermatozoa to move about, which could have positive therapeutic effects.

Global DNA methylation level in spermatozoa is not associated with ICSI fertilization outcome and embryo quality in donor oocyte programme

Epigenetic mechanisms, including DNA methylation and histone modifications, govern chromatin arrangement in sperm, enhancing motility and safeguarding DNA integrity for accurate epigenetic inheritance. Abnormal methylation is linked to poor sperm quality and fertility issues, underscoring the need to study sperm DNA methylation and its impact on sperm function and embryo development in assisted reproductive technology. In this study, processed spermatozoa from 75 normozoospermic and 15 abnormal ejaculates were examined for sperm global DNA methylation levels using a colourimetric absorbance method. Although semen characteristics were poor in abnormal ejaculates, no significant correlation was found between sperm global DNA methylation levels and sperm characteristics in either normozoospermic or abnormal cohorts. However, mean global DNA methylation levels were significantly lowered in abnormal sperm samples compared to normozoospermic samples (p < 0.05). Furthermore, injecting spermatozoa from these patients (N = 50) into donor oocytes did not show a significant relationship between sperm global DNA methylation and embryo developmental competence. These findings highlight the limitation of sperm global DNA methylation as a biomarker for embryo development and quality.

Pharmacological inhibition of KSper impairs flagellar pH homeostasis of human spermatozoa.

Sperm-specific potassium channel (KSper) comprised of pore-forming subunit SLO3 and auxiliary subunit LRRC52 is of importance for sperm fertility. The deficiency of KSper in both mice and humans resulted in severe impairments of sperm functions including sperm hyperactivity and acrosome reaction. Previous reports suggested that mouse KSper modulated sperm function possibly by affecting sperm intracellular pH (pHi). However, the precise signaling mechanism of human KSper (hKSper) on the regulation of sperm functions was largely unclear. To explore the regulatory role of hKSper on sperm flagellar pHi. More than 50 sperm donors were recruited during a period of 1 year. As reported in our previous work, we quantitatively measured flagellar pHi by employing a single-cell pH fluorescent recording on human spermatozoa loaded with pH indicator pHrodo Red. Three different hKSper antagonists including clofilium, quinidine, and a polyclonal antibody of LRRC52 (LID1) were utilized to evaluate the effect of hKSper inhibition on sperm flagellar pHi. Given the predominant role of hKSper on the regulation of membrane potential (Em), we first detected a considerable depolarization (about 25-30mV) of Em evoked by clofilium and quinidine. Subsequently, it was shown that flagellar pHi values of human spermatozoa were significantly decreased by the treatment of clofilium (50 µM, from 7.13±0.11 to 6.43±0.12), quinidine (500 µM, from 7.00±0.11 to 6.64±0.08) and LID1 (20 µg/mL, from 6.98±0.16 to 6.67±0.22). Moreover, we found that when human spermatozoa were pre-incubated with a high K+ solution (135mM), both the depolarization of Em and the acidification of flagellar pHi evoked by clofilium and quinidine were abolished. In addition, we found that extracellular substitution of N-methyl-D-glucamine for Na+ abolished pHi acidification induced by hKSper inhibition. Our results demonstrate that hKSper inhibition evokes flagellar pHi acidification of human spermatozoa, suggesting that flagellar pHi maintenance is an important signaling mechanism of hKSper on the regulation of sperm functions.

Role of Rab proteins in PFOA-induced changes in boar sperm motility and capacitation

Perfluorooctanoic acid (PFOA), a pervasive environmental contaminant, elicits adverse effects on sperm functions, including sperm motility and capacitation status. However, the specific mechanisms by which PFOA disrupts sperm functions during capacitation remain poorly elucidated. Therefore, this study aimed to investigate the molecular mechanisms underlying the PFOA-induced inhibition of sperm motility and capacitation in boar spermatozoa by focusing on Ras-related (Rab) proteins, which regulate membrane trafficking and play key roles in male sperm development, acrosome formation, and the acrosome reaction. Results showed significant reductions in sperm motility and various kinematic parameters following PFOA exposure. Correlation analysis revealed that Rab14 was positively correlated with dance mean (DNM) and negatively correlated with wobble (WOB), indicating that PFOA might affect sperm motility through Rab14 and potentially lead to reduced pregnancy rates. Differences in Rab25 were positively correlated with differences in total motility (MOT), progressive motility (PRG), linearity (LIN), and mean angular displacement (MAD), suggesting that PFOA might influence sperm motility by altering Rab25. Differences in Rab34 were positively correlated with differences in acrosome-reacted spermatozoa, implicating its role in the acrosome reaction. These findings provided insights into the molecular mechanism of PFOA-induced reproductive toxicity and highlighted the function of Rab proteins as biomarkers for the assessment of the effects of similar environmental toxins on male fertility.

In Vitro Spermatogenesis on Human Decellularized Testicular Matrix Plates Following Exosome Treatment in a Dynamic Culture System.

Testicular tissue engineering for in vitro spermatogenesis aims to restore fertility, focusing on challenges like efficiency, ethical concerns, and the need for a deeper biological understanding. The use of decellularized scaffolds led to better cell seeding and differentiation, and exosomes led to enhanced spermatogenesis. Also, the dynamic culture systems are being explored to replicate in vivo conditions more accurately. In this study, we aimed to utilize a perfusion mini-bioreactor for the dynamic culture of mouse spermatogonial stem cells on decellularized testicular matrix plates supplemented with exosomes. Our goal was to assess the progression of the spermatogenesis process through histological, immunohistochemical, and molecular analyses over four weeks. Human testicular tissues were decellularized using 1% sodium dodecyl sulfate and were then fabricated into thin plates using a cryostat. Sertoli and spermatogonial stem cells were isolated from neonate mouse testis and seeded onto the decellularized testicular matrix plates. A mini-perfusion bioreactor was employed to create dynamic culture conditions. Also, MSCs-derived exosomes were introduced to the culture medium, alone or in combination with a spermatogenic medium containing numerous chemical factors. The histological, IHC, and molecular analyses were performed at the end of the experiment. Our decellularization procedure successfully preserved the ECM components, while eliminating native cells. The isolated cells expressed PLZF and VIMENTIN markers, confirming the presence of SSCs and Sertoli cells. The seeded scaffolds exhibited proper homing, viability, proliferation, and differentiation of the cells towards in vitro spermatogenesis. Also, exosome treatment is capable of enhancing the spermatogenic potential of SSCs. Our findings indicate that the dynamic culture system significantly promoted the proliferation and differentiation of SSCs into mature spermatozoa. The use of exosomes further enhanced these effects, as evidenced by improved cellular viability, reduced apoptosis, and advanced spermatogenesis to the elongated spermatid stage. The combined treatment of exosomes and spermatogenic medium showed a synergistic effect, yielding superior outcomes in terms of sperm cell maturity and functionality. This study underscores the potential of combining decellularized testicular matrices with exosome therapy in a dynamic culture set up to advance the field of reproductive biology and fertility restoration.

Impact of Reductive Stress on Human Infertility: Underlying Mechanisms and Perspectives

Antioxidants have a well-established effect on general health and are essential in preventing oxidative damage to cells by scavenging free radicals. Free radicals are thought to be neutralized by these substances, which include polyphenols, β-carotene, and vitamins C and E, reducing cellular damage. On the other hand, recent data indicates that consuming excessive amounts of antioxidants may have side effects. Apoptosis and cell signaling are two beneficial physiological processes that are affected by excessive supplementation. Other negative effects include paradoxical enhancement of oxidative stress and unbalanced cellular redox potential. Overdosing on particular antioxidants has been associated with increased medication interactions, cancer progression, and fatality risks. Additionally, the complex impacts they may have on fertility might be both useful and adverse, depending on the quantity and duration of usage. This review delves into the dual role of antioxidants and emphasizes the importance of employing antioxidants in moderation. Antioxidant overconsumption may disrupt the oxidative balance necessary for normal sperm and oocyte function, which is one of the potential negative effects of antioxidants on fertility in both males and females that are also investigated. Although modest usage of antioxidants is generally safe and useful, high levels of antioxidants can upset hormonal balance, impair sperm motility, and negatively impact the outcomes of assisted reproductive technologies (ART). The findings emphasize the need to use antioxidant supplements in a balanced way, the importance of further research to optimize their use in fertility treatments, and the importance of supporting reproductive health to avoid adverse effects.

Cryopreservation of yellowtail kingfish (Seriola lalandi) sperm: Effects on metabolic enzymes and sperm quality

This study aimed to determine the effects of cryopreservation on sperm functionality and assess post-thaw sperm quality in yellowtail kingfish (Seriola lalandi). Milt designated for cryopreservation was frozen in a solution containing Cortland® medium +1.3 M Me2SO + 0.2 M glucose +2 % BSA in a 1:3 ratio (milt:cryoprotectant). Fresh milt was used as the control. The samples were frozen in 0.5 mL straws using a programmable freezing system. After thawing, the samples were evaluated for motility using Computer-Assisted Sperm Analysis (CASA) with a phase-contrast optical microscope. Additionally, flow cytometry and a multimodal reader were employed to assess various sperm parameters, including plasma membrane integrity (SYBR-14/PI), mitochondrial membrane potential (∆ΨMMit: JC-1), DNA fragmentation (TUNEL), ATP content (CellTiter-Glo®), intracellular calcium levels (Fluo-4 AM), lipid peroxidation (LPO), intracellular superoxide (DHE/SYTOX) and glutathione (GSH/GSSG) production, activities of glutathione peroxidase (GPx), catalase (CAT) and 8-oxoguanine glycosylase (OGG1), and detection of 8-hydroxydeoxyguanosine (8-OHdG). In cryopreserved milt, significant differences in motility (64.14 ± 2.2 %), plasma membrane integrity (61.5 ± 2.7 %), mitochondrial membrane potential (55.4 ± 2.4 %), DNA fragmentation (6.7 ± 1.1 %), ATP content (7.5 ± 1.5 nmol/109 spermatozoa), and intracellular calcium levels (59.8 ± 2.5 nM/109 spermatozoa) were observed compared to fresh milt. Differences were also noted in LPO, O2−i, GPx, GSH/GSSG, CAT, 8-OHdG and OGG1 between the cryopreserved and control groups. Additionally, correlations between sperm quality parameters, oxidative stress markers, and enzyme activity in cryopreserved spermatozoa were determined. These results suggest that cryopreservation induces significant alterations derived from oxidative damage in the functionality of yellowtail kingfish spermatozoa, and thus further research is necessary to explore the need for using antioxidants in the cryopreservation process of sperm from this species.

Analysis of stallion spermatozoa metabolism using Agilent Seahorse XFp Technology

Sperm metabolism consists of a sophisticated network of biochemical reactions and varies between species, resulting in different metabolic strategies for ATP production to maintain sperm functionality. ATP can be produced through glycolysis or in the mitochondria by oxidative phosphorylation (OXPHOS). Since OXPHOS is the predominant metabolic pathway in horses spermatozoa, various assessments of mitochondrial activity are used to evaluate fertility, utilizing techniques such as fluorescent probes analysed via microscopy or flow cytometry, and polarographic electrode assays to measure current flow in response to an applied voltage. Though, these methods are limited by low throughput, as they assess mitochondrial activity at a single time point under a specific treatment condition. This study explores, for the first time, the application of the Agilent Seahorse XFp Technology to evaluate metabolism in stallion spermatozoa. This method enables real-time measurement of cellular metabolism across multiple samples or experimental conditions simultaneously. Ejaculates from eight different stallions were collected, and pools were prepared from three of them. Sperm viability and mitochondrial activity were evaluated by fluorescence microscopy, sperm motility by a computer-assisted sperm analysis system, and sperm metabolism was analysed via the Seahorse XFp analyser. Results confirmed a preference for OXPHOS over glycolysis in ATP production in stallion sperm, with mitochondria contributing significantly to total ATP generation. The Seahorse XFp Technology proved effective in evaluating equine sperm bioenergetics, offering insights into metabolic pathways critical for sperm function. In conclusion, this technology grants a new method for high-throughput analysis of sperm metabolism and quality, which could be applied to future reproductive studies in male equine fertility.

Treatment of human sperm with GYY4137 increases sperm motility and resistance to oxidative stress.

Hydrogen sulfide (H2S) has been shown to play a significant role in oxidative stress across various tissues and cells; however, its role in sperm function remains poorly understood. This study aimed to investigate the protective effect of GYY4137, a slow-releasing H2S compound, on sperm damage induced by H2O2. We assessed the effects of GYY4137 on motility, viability, lipid peroxidation and caspase-3 activity in human spermatozoa in vitro following oxidative damage mediated by H2O2. Spermatozoa from 25 healthy men were selected using a density gradient centrifugation method and cultured in the presence or absence of 10 μM H2O2, followed by incubation with varying concentrations of GYY4137 (0.625-2.5 μM). After 24 h of incubation, sperm motility, viability, lipid peroxidation, and caspase-3 activity were evaluated. The results indicated that H2O2 adversely affected sperm parameters, reducing motility and viability, while increasing oxidative stress, as evidenced by elevated lipid peroxidation and caspase-3 activity. GYY4137 provided dose-dependent protection against H2O2-induced oxidative stress (OS). We concluded that supplementation with GYY4137 may offer antioxidant protection during in vitro sperm preparation for assisted reproductive technology.

The regulation role of calcium channels in mammalian sperm function: a narrative review with a focus on humans and mice.

Mammalian sperm are characterized as specialized cells, as their transcriptional and translational processes are largely inactive. Emerging researches indicate that Ca2+ serves as a crucial second messenger in the modulation of various sperm physiological processes, such as capacitation, hyperactivation, and the acrosome reaction. Specifically, sperm-specific calcium channels, including CatSper, voltage-gated calcium channels (VGCCs), store-operated calcium channels (SOCCs), and cyclic nucleotide-gated (CNG) channels, are implicated in the regulation of calcium signaling in mammalian sperm. Calcium stores located in the sperm acrosomes, along with the IP3 receptors in the neck of the redundant nuclear envelope and the mitochondria in the tail, play significant roles in modulating intracellular Ca2+ levels in sperm. However, the functions and mechanisms of these calcium channels in modulating mammalian sperm physiological functions have not yet been well elucidated. Therefore, by focusing on humans and mice, this study aims to provide a comprehensive review of the current advancements in research regarding the roles of calcium signaling and associated calcium channels in regulating sperm function. This endeavor seeks to enhance the understanding of calcium signaling in sperm regulation and to facilitate the development of drugs for the treatment of infertility or as non-hormonal male contraceptives.

Microplastics are detected in bull epididymal sperm and polystyrene microparticles impair sperm fertilization.

An increase in global infertility has coincided with the accumulation of microplastics (MPs) in the environment. This trend is particularly troubling because only 10% of male infertility cases can be attributed to identifiable causes, leaving a knowledge gap in our understanding of their underlying factors. To bridge this, it is important to explore the connection between the accumulation of MPs and the observed decline in male fertility. We assessed the presence of microplastics in epididymal sperm from bulls and used it as baseline concentrations for sperm exposure. MPs were detected in all epidydimal sperm (ES) samples, with a mean concentration of 0.37μgmL-1. Next, to investigate the effects of MPs on fertility, bovine sperm was exposed to three different concentrations of a mixture of 1.1, 0.5, and 0.3μm polystyrene (PS) beads: (1) 0.7μgmL-1, blood concentration of PS in cows (bPS); (2) 0.37μgmL-1, based on the concentration of total MPs found in ES (esMP); and (3) 0.026μgmL-1, based on the concentration of PS found in ES (esPS). All sperm samples incubated with PS exhibited reduced motility compared with the control at 0.5h. However, PS exposure did not affect acrosome integrity or induced oxidative stress. Embryos produced from sperm exposed to PS had reduced blastocyst rates, in addition to increased ROS formation and apoptosis. By employing physiological exposure, this research provided evidence of MPs in bovine epididymal sperm and demonstrated the detrimental effect of PS on sperm functionality.

The effect of myo-inositol antioxidant activity on human sperm parameters and DNA damage in ultra-rapid and conventional freezing methods

Male fertility preservation is still challenged by cell damage induced during sperm cryopreservation and impaired sperm structure and function. Sperm ultra-rapid freezing, despite a higher protective effect compared to conventional freezing method, is still associated with suboptimal sperm cryosurvival and needs to be modified to increase its efficiency in sperm protection. Sperm freezing media supplemented with antioxidants can improve sperm parameters following freezing-warming process. In this study, we aimed to investigate the effect of employing ultra-rapid freezing and myo-inositol on sperm cryosurvival. Thirty semen samples with normal sperm parameters were collected and each one was divided into four portions to cryopreserve by conventional freezing, ultra-rapid freezing, conventional freezing + myo-inositol 2 mg/ml, and ultra-rapid freezing + myo-inositol 2 mg/ml. Sperm samples warmed after at least 24 h of freezing and sperm cryosurvival were analyzed by evaluation of sperm motility, viability, morphology and DNA fragmentation index (DFI). Freezing method had a significant influence on post-thaw sperm DFI and morphology (p < 0.05) and the interaction between freezing method and antioxidant supplementation significantly affected sperm morphology (p < 0.05). The highest percentage of sperm normal morphology and minimal DFI was achieved using ultra-rapid freezing supplemented by myo-inositol antioxidant compared to other groups (P < 0.05). The highest sperm DNA damage after freezing-warming was observed following the conventional freezing method. In conclusion, sperm freezing method was identified as factor strongly influencing sperm DFI and morphology after thawing/warming. Sperm samples can be rapidly frozen using the modified freezing media supplemented by myo-inositol without impacting sperm DNA and morphology.

Effect of Oxidative Stress on Sperm TERRA Expression and Chromatin Function in Infertile Males: A Preliminary Finding

Telomeres are vital for cellular function. Oxidative stress is detrimental to telomere function as it causes telomere attrition. Increase in oxidative stress has a negative impact on sperm telomere and chromatin integrity and therefore, on sperm function and male fertility. TERRA, a long non-coding RNA is a fundamental component required for telomere length homeostasis. Aim: The present work aimed to study the effect of H2O2 induced oxidative stress on sperm TERRA expression and chromatin function. Methodology: 5 infertile and 5 fertile males were selected for the study. Baseline semen analysis was done as per WHO manual 2021. Sperm TERRA expression was assessed by real time qPCR. To assess chromatin function nuclear chromatin decondensation test was done. Results: Study showed that 500µM H2O2 increased TERRA expression as compared to untreated groups and the difference was found to be statistically significant. As compared to controls, cases showed more fold increase in TERRA expression. When treated samples of control and cases were compared, cases had more TERRA expression than controls however, the baseline TERRA expressions in untreated samples were less in cases as compared to controls. Both comparisons yielded statistically nonsignificant results. When nuclear chromatin decondensation was compared before and after H2O2 treatment, the percentage of sperm head decondensation decreased in both cases and controls. When treated and untreated samples of cases and controls were compared, the difference was statistically nonsignificant. Conclusion: Oxidative stress led to an increase in the expression of sperm TERRA and had a negative impact on nuclear chromatin decondensation. Similar studies with greater sample size can provide much more insight into TERRA mediated regulation of telomere function and chromatin integrity and how it is affected by oxidative stress.

Sperm histone modifications may predict success in human assisted reproduction: a pilot study.

Currently, assisted reproduction clinics employ various sperm selection techniques to identify the best sperm for fertilization. However, these techniques may not assess crucial sperm traits that can substantially impact embryonic quality. To address this, we propose analyzing diverse histone modifications as potential markers of sperm functionality and success in assisted reproduction techniques. Cross-sectional pilot study including infertile male patients attending an infertility clinic in CABA, Argentina between April and August 2019 was performed. We used immunofluorescence techniques to evaluate post-translational modifications of histones in sperm and established correlations with in vitro fertilization outcome and embryo quality. Our findings indicate a negative correlation between H3K4me3 and H3K4me2 marks and fertilization rate and showed a positive correlation of this parameter with H3K9me mark. In addition, there was a positive correlation between H3K27me3 and good embryo quality. This pilot study proposes a non-invasive strategy to predict embryo quality by analyzing spermatozoa prior to fertilization. The assessment of histone post-translational modifications in sperm samples could provide useful information for the recognition of epigenetic marks that could predict the health of the embryo of an assisted fertilization treatment.

Real-Time Imaging of Calcium Dynamics in Human Sperm After Precise Single-Cell Stimulation.

Calcium signaling is a critical regulator of sperm activation and function during the processes of capacitation and fertilization. Here, we describe a combined method for calcium imaging of single, live human sperm in response to stimuli administered with a precisely targeted delivery technique. This protocol is an adaptation of techniques developed for studies of murine sperm [1, 2], and enables real-time monitoring of human sperm calcium dynamics with high spatiotemporal resolution and concurrent detection of acrosome exocytosis (AE), a functional endpoint of sperm capacitation and requirement for physiological fertilization.The described imaging technique provides a valuable tool for exploration of calcium regulation in human sperm, which is essential to answer important questions and knowledge gaps regarding the link between calcium dynamics, AE, and fertilization. The versatility of this technique can be amplified through use of various indicator dyes or integration with pharmacological strategies such as pre-treating sperm with inhibitors or activators targeting specific receptors, channels, or intracellular signaling pathways of interest. Beyond fundamental inquiries into sperm physiology, this method can also be applied to assess the impact of potential contraceptive compounds on calcium signaling, AE, and membrane integrity.

Voltage-Dependent Anion Channels in Male Reproductive Cells: Players in Healthy Fertility?

Male infertility affects nearly 50% of infertile couples, with various underlying causes, including endocrine disorders, testicular defects, and environmental factors. Spermatozoa rely on mitochondrial oxidative metabolism for motility and fertilization, with mitochondria playing a crucial role in sperm energy production, calcium regulation, and redox balance. Voltage-dependent anion channels (VDACs), located on the outer mitochondrial membrane, regulate energy and metabolite exchange, which are essential for sperm function. This review offers an updated analysis of VDACs in the male reproductive system, summarizing recent advances in understanding their expression patterns, molecular functions, and regulatory mechanisms. Although VDACs have been widely studied in other tissues, their specific roles in male reproductive physiology still remain underexplored. Special attention is given to the involvement of VDAC2/3 isoforms, which may influence mitochondrial function in sperm cells and could be implicated in male fertility disorders. This update provides a comprehensive framework for future research in reproductive biology, underscoring the significance of VDACs as a molecular link between mitochondrial function and male fertility.

Neonatal undernutrition induced by litter size expansion alters testicular parameters in adult Wistar rats.

Several models of maternal undernutrition reveal impairment of testicular development and compromise spermatogenesis in male offspring. The expansion of the litter size model, valuable for studying the impact of undernutrition on early development, has not yet been used to evaluate the consequences of early undernutrition in the adult male reproductive system. For this purpose, pups were raised in either normal litter (ten pups/dam) or large litter (LL; sixteen pups/dam). On postnatal day 90, sexual behaviour was evaluated or blood, adipose and reproductive tissues were collected for biochemical, histological and morphological analysis. Adult LL animals were lighter and thinner than controls. They showed increased food intake, but decrease of retroperitoneal white adipose tissue weight, glycaemia after oral glucose overload and plasma concentration of cholesterol. Reproductive organ weights were not altered by undernutrition, but histopathological analysis revealed an increased number of abnormal seminiferous tubules and number of immature spermatids in the tubular lumen of LL animals. These animals also showed reduction in total spermatic reserve and daily sperm production in the testes. Undernutrition decreased the number of Sertoli cells, and testosterone production was increased in the LL group. Mitochondrial activity of spermatozoa remained unchanged between experimental groups, suggesting no significant impact on the energy-related processes associated with sperm function. All animals from both experimental groups were considered sexually competent, with no significant difference in the parameters of sexual behaviour. We conclude that neonatal undernutrition induces histological and physiological testicular changes, without altering sperm quality and sexual behaviour of animals.

Bleomycin in vitro exposure decreases markers of human male gamete competence

To investigate the in vitro impact of bleomycin on human sperm DNA integrity, functionality, and morphology, with the aim of elucidating the underlying mechanism and anticipating potential repercussions on patients' reproductive function. Controlled laboratory-based in vitro investigation. Surplus human ejaculate donated for research by 45 reproductive-age participants exhibiting normozoospermic sperm parameters following clinical semen analysis. None of the participants had received a cancer diagnosis or undergone radiotherapy, chemotherapy, or both. After clinical semen analysis, sperm samples were centrifuged, diluted in sperm preparation medium (SPM), and exposed to bleomycin (100 μg/mL) for two hours at 37ºC in a humidified incubator with 5% CO2. In vitro human sperm competence was evaluated by comparing raw sperm, sperm incubated with SPM, and sperm exposed to bleomycin. Competence indicators included sperm motility, vitality, DNA and acrosome integrity, and mitochondrial membrane potential. Transmisson electron microscopy was employed to correlate the ultrastructural morphological findings with functional assays. Exposure to bleomycin for two hours in vitro significantly (P < 0.001) decreased sperm vitality, motility, and chromatin condensation compared to raw and control sperm. It also significantly (P < 0.001) increased sperm DNA fragmentation and the proportion of sperm with low mitochondrial membrane potential. Additionally, bleomycin significantly (P < 0.05) retarded the acrosomal response compared to control but did not affect the formation of intracellular and extracellular reactive oxygen species. Bleomycin-induced ultrastructural morphological changes supported the detected functional alterations. Bleomycin negatively impacts male gamete competency in humans. Healthcare professionals should vigilantly monitor and further investigate the gonadotoxicity effects of bleomycin, in addition to its recognized lung toxicity. Meanwhile, it is recommended that cancer patients undergoing bleomycin-containing chemotherapy regimens receive guidance on fertility preservation strategies.

Influence of Dietary Fatty Acids on Fish Sperm Tolerance to Cryopreservation

ABSTRACTTo meet growing fish demand, aquaculture must develop sustainable, cost‐effective, and high‐performance fish feed formulations for industry advancement. Plant‐based lipids are a viable alternative to forage fish ingredients in finfish diets. However, most plant‐derived lipids lack long‐chain polyunsaturated fatty acids (LC‐PUFA) like EPA and DHA, which are nutritionally essential for fish. This review summarizes recent findings on the nutritional aspects of fish brood stock and sperm quality, emphasizing the effects of fatty acids on reproductive outcomes. Studies indicate that substituting fish oil with plant oils alters the fatty acid profiles in fish tissues and gametes, potentially compromising sperm quality, cryopreservation success, and progeny viability. Sperm membrane lipids, including EPA, DHA, and ARA, are vital for motility and fertilization rates post‐thaw. Additionally, cryopreservation has been linked to molecular and epigenetic alterations in sperm, which may negatively affect offspring quality. The role of mRNA and noncoding RNA in regulating sperm function and embryonic development is well known, and thus any damage to these molecules as a consequence of cryopreservation might induce long‐term effects on offspring. Proteomic analyses reveal that cryopreservation can lead to significant protein loss in sperm, diminishing their functional and fertilization capabilities. Optimizing cryoprotectant protocols and freezing techniques is crucial to reduce damage, while dietary fatty acids play a key role in preserving sperm quality during cryopreservation stress. Further research with omic technologies is crucial to fully understand sperm tolerance to cryopreservation, which will improve reproductive outcomes and enhance sustainability and quality in aquaculture.

The Effect of Cyperus esculentus on Sperm Function Parameters in Prepubertal Mice as a Model for Human

The objective of this work was to study the effect of oral administration of Cyperus esculentus (CE) and its alcoholic extract on sperm function parameters in prepubertal mice as a model for human .The animals were divided into three groups each contains 6 animals .Group 1 was treated with 150 mg/ kg body weight /day of crude CE, group 2 was treated with same dose of alcohol extract of CE and group 3 regarded as control throughout six weeks period. The results showed a significant (p&gt; 0.05) increase in the mean of sperm concentration ,sperm motility percent and progressive sperm motility between treated groups and control . There was no differences among groups in the mean of sperm normal morphology and sperm viability . No significant differences was recorded in the mean of body weight among groups throughout the study. The results revealed that the administration of Cyperus esculentus may enhance certain sperm characters in prepubertal mice without affecting body weight.