Sperm Activation Research Articles

SPSL1 is essential for spermatophore formation and sperm activation in Spodoptera frugiperda.

The reproductive process in various species has undergone evolutionary adaptations at both the physiological and molecular levels, playing a significant role in maintaining their populations. In lepidopteran insects, the spermatophore is a unique structure formed in the female reproductive system, in which sperm storage and activation take place. It is known that the formation of the spermatophore is regulated by seminal fluid proteins derived from males. However, studies investigating the genetic mechanisms behind spermatophore formation in lepidopterans have been limited. In this study, our focus was on SPSL1, a gene that encodes a trypsin-type seminal fluid protein in Spodoptera frugiperda, a pest species with global invasive tendencies. Our findings revealed that SPSL1 expression was predominantly observed in the male reproductive tracts, and the disruption of this gene resulted in male sterility. Surprisingly, fluorescence analysis indicated that the absence of SPSL1 did not affect spermatogenesis or sperm migration within the male reproductive system. However, when females mated with SPSL1-mutant males, several defects were observed. These included disruptions in spermatophore formation, sperm activation in the copulatory bursae, and sperm migration into the spermathecae. Additionally, mass spectrometry analysis highlighted reduced levels of energy-related metabolites, suggesting that SPSL1 plays an essential role in promoting hydrolysis reactions during copulation. Consequently, our study demonstrates that SPSL1 is crucial for male fertility due to its functions in spermatophore formation and sperm activation. This research provides valuable insights into the genetic factors underlying reproductive processes in lepidopteran insects and sheds light on potential strategies for controlling invasive pest populations.

Combination Therapy with Enalapril and Paricalcitol Ameliorates Streptozotocin Diabetes-Induced Testicular Dysfunction in Rats via Mitigation of Inflammation, Apoptosis, and Oxidative Stress.

As the impacts of diabetes-induced reproductive damage are now evident in young people, we are now in urgent need to devise new ways to protect and enhance the reproductive health of diabetic people. The present study aimed to evaluate the protective effects of enalapril (an ACE inhibitor) and paricalcitol (a vitamin D analog), individually or in combination, on streptozotocin (STZ)-diabetes-induced testicular dysfunction in rats and to identify the possible mechanisms for this protection. This study was carried out on 50 male Sprague-Dawley rats; 10 normal rats were allocated as a non-diabetic control group. A total of 40 rats developed diabetes after receiving a single dose of STZ; then, the diabetic rats were divided into four groups of equivalent numbers assigned as diabetic control, enalapril-treated, paricalcitol-treated, and combined enalapril-and-paricalcitol-treated groups. The effects of mono and combined therapy with paricalcitol and enalapril on testicular functions, sperm activity, glycemic state oxidative stress, and inflammatory parameters, as well as histopathological examinations, were assessed in comparison with the normal and diabetic control rats. As a result of diabetes induction, epididymal sperm count, sperm motility, serum levels of testosterone, follicle-stimulating hormone (FSH) as well as luteinizing hormone (LH), and the antioxidant enzyme activities, were significantly decreased, while abnormal sperm (%), insulin resistance, nitric oxide (NO), malondialdehyde (MDA), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were significantly increased, along with severe distortion of the testicular structure. Interestingly, treatment with paricalcitol and enalapril, either alone or in combination, significantly improved the sperm parameters, increased antioxidant enzyme activities in addition to serum levels of testosterone, FSH, and LH, reduced insulin resistance, IL-6, and TNF-α levels, and finally ameliorated the diabetes-induced testicular oxidative stress and histopathological damage, with somewhat superior effect for paricalcitol monotherapy and combined therapy with both drugs compared to monotherapy with enalapril alone. Monotherapy with paricalcitol and its combination therapy with enalapril has a somewhat superior effect in improving diabetes-induced testicular dysfunction (most probably as a result of their hypoglycemic, antioxidant, anti-inflammatory, and anti-apoptotic properties) compared with monotherapy with enalapril alone in male rats, recommending a synergistic impact of both drugs.

Effect of pH, osmolality, Na+ and K+ on spermatozoa motility of the orange-spotted grouper, Epinephelus coioides

ABSTRACT The present study investigated the biosensitivity of Epinephelus coioides spermatozoa under various ionic and osmolality conditions. Activation and sensitivity of sperm started at different pH (7, 7.5, 8, 8.5, and 9) level. The highest motility duration was obtained in pH 8.5 with a significant difference to other groups (P < .001). The E. coioides spermatozoa exposed to 600 mM NaCl solution, 300 mM KCl (in the medium containing 200 mM sucrose), and sucrose solutions with 1100 mosmol/kg osmolality had significantly higher duration of motility than those exposed to solutions with the lower amount of NaCl, KCl and osmolality. Any amount greater than 600 mM NaCl, 300 mM KCl and 1100 mosmol/kg osmolality showed a remarkable reduction in the motility of spermatozoa. Results also showed that the amount of 100 and 450 mM K+, and osmolality lower than 400 mosmol/kg had a significant inhibitory effect and prevented the activity of spermatozoa. The findings of this study showed that changes in K+, Na+ and osmolality of the external solutions affects the spermatozoa and its motility. The potential use of these ions could be applicable for the improvement of the spermatozoa quality of E. coioides during artificial reproduction.

Erythritol attenuates testicular dysfunction in diabetic rat via suppression of oxidative stress, inflammation and apoptosis

Hyperglycemia –induced oxidative stress and inflammation have been closely associated with diabetes complications including testicular dysfunction. Conversely, reducing blood glucose and/or use of antioxidant have been associated with reduced diabetes complications. The present study investigated the effect of erythritol (which has both antioxidant and blood glucose lowering function) on diabetes –induced testicular dysfunction in rats. Thirty male Wistar rats (170–200g) were randomly divided into 5 groups: 1) control; 2) erythritol; 3) diabetic; 4) diabetic + erythritol 1000 mg/kg; and 5) diabetic + metformin 300 mg/kg. After 8 weeks of treatment period, blood sample, testes and epididymis were collected for reproductive hormones, biochemical and histological examinations, and sperm analysis respectively. There was a significant (p < 0.05) decrease in sperm count, sperm motility, sperm morphology and serum reproductive hormones (Follicle stimulating hormone (FSH), Leutinizing hormone (LH), testosterone and gonadotropin releasing hormone (GnRH)) of diabetes rat compared to control. Also, diabetes rat showed increase in sperm and testicular malonaldehyde (MDA) and decrease in sperm and testicular superoxide dismutase (SOD) activity and glutathione (GSH) level. Further, diabetes rat showed reduced testicular weight, decreased testicular 17β-HSD and 3β-HSD activity and testicular histo-architectural alteration which were accompanied by decrease testicular vascular endothelial growth factor (VEGF) and concomitant increase in testicular myeloperoxidase activity and level of caspase 3. The present results indicates that induction of diabetes in rat causes reduction in the level of reproductive hormones (Testosterone, LH and FSH) as well as sperm and testicular oxidative stress causing abnormal sperm parameters, and biochemical and histo-architectural alterations in the testes of rats. In addition, the present results suggest that erythritol administration reduced blood glucose and ameliorated hyperglycemia –induced oxidative stress –mediated alterations in both sperm and testes of diabetes rat. Further, the present study suggests that erythritol improved testicular oxidative stress, inflammation and apoptosis by up-regulating VEGF.

Early-life long-term ibuprofen exposure reduces reproductive capacity involved in spermatogenesis impairment and associated with the transcription factor DAF-5 in Caenorhabditis elegans

Pharmaceuticals and personal care products (PPCPs) are emerging environmental contaminants and have raised significant concern due to their potential adverse impact on the environment. Ibuprofen is one of the most extensively used non-steroidal anti-inflammatory drugs (NSAIDs) and is also considered an environmental contaminant. The negative impact of ibuprofen on non-target organisms has been documented; however, the molecular mechanisms behind its reproductive toxicity remain unclear. We investigated the impact of early-life long-term ibuprofen exposure on reproductive capacity and its involvement of spermiogenesis in the non-target model organism Caenorhabditis elegans. Hermaphrodites were exposed to various ibuprofen concentrations (0.1, 1, 10, and 100 mg/L), resulting in a dose-dependent inhibition of reproduction. In addition, the lowest observed adverse effect concentration (LOAEC) for ibuprofen exposure on the total brood size of C. elegans was 0.1 mg/L, a concentration that falls within the environmentally relevant range for ibuprofen. Outcross progeny assays revealed a significant 47% reduction in total brood size for larval males (him-5) exposed to ibuprofen, while females (fog-2) exhibited only a minor effect. We found that early-life long-term ibuprofen exposure impairs spermatogenesis. The number of mitotic cells significantly reduced by 31%. The rate of sperm malformation in exposed males was 63%, much higher than in unexposed males (11%). Additionally, the percentage of sperm activation decreased from 89% to 39% in ibuprofen-exposed worms. Mechanistic insights indicated that ibuprofen downregulated mRNA levels of genes related to spermatogenesis and DAF-7/TGF-β signaling. RNAi assays provided evidence for the crucial role of the transcription factor DAF-5 in mediating the spermatogenesis impairment by ibuprofen. Our study provides insight into the environmental impacts of pharmaceutical contaminants, such as ibuprofen, on both male and female reproductive systems to safeguard environmental health.

Factors determining sperm fertility

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

Effect of Zinc Supplementation to Diluent Boar Semen on Sperm Characteristics and Activity of Antioxidant Enzymes

Background. Zinc is essential for male reproductive function as it is necessary for spermatozoa maturation, capacitation, acrosome reaction, and fertilization. It has been established that the use of chelates compounds of metals with amino acids, nucleotides, peptides, and carbohydrates is not only more effective but also economically justified. Objective. The aim of this study was to evaluate and compare the effect of different concentrations of Zn glutamic amino acid chelate (ZnGlu) on boar spermatozoa motility, viability parameters, and prooxidant-antioxidant homeostasis during in vitro incubation. Methods. Freshly ejaculated boar semen, after the addition of "Ecosperm" diluent, was divided into groups: a control group and three experimental groups. Zinc glutamate was supplemented to the experimental samples at concentrations of 1.0, 2.0, and 5.0 µg/ml, respectively. Semen samples were stored at 18 °C for 4 days. Every 24 hours, sperm motility and viability and the antioxidant status were assessed by the level of diene con­jugates (DC), concentration of malondialdehydere (MDA), and the activities of catalase and superoxide dismutase. Results. It was observed that boar sperm motility decreased with the extension of storage time, while the addition of 2.0 and 5.0 µg ZnGlu significantly improved sperm total motility and the percentage of vitality spermatozoa during 48–96 hours of incubation. Our studies also demonstrate that ZnGlu possess a protective effect in alleviating oxidative stress in boar sperm in vitro. The addition of ZnGlu significantly reduced the content of MDA and DC in ejaculate samples in all experimental groups during incubation, compared to the control group. Moreover, the activity of superoxide dismutase and catalase increased after adding ZnGlu to the boar semen, especially at concentrations of 2.0 and 5.0 mg/ml (P &lt; 0.05). Conclusions. These data demonstrate that the supplementation of zinc glutamate enhances the antioxidant defence system of sperm and improves quality of boar semen in vitro.

A Hand-Held Platform for Boar Sperm Viability Diagnosis Based on Smartphone.

The swine fever virus seriously affects pork production, and to improve pork production, pig breeding efficiency needs to be improved, and the detection of boar sperm activity is an important part of the pig breeding process. Traditional laboratory testing methods rely on bulky testing equipment, such as phase-contrast microscopes, high-speed cameras, and computers, which limit the testing scenarios. To solve the above problems, in this paper, a microfluidic chip was designed to simulate sperm in the oviduct with a channel thickness of 20 um, which can only accommodate sperm for two-dimensional movement. A miniature microscope system which can be used in combination with a smartphone is designed that is only the size of the palm of the hand and has a magnification of about 38 times. An intelligent diagnostic app was developed using Java language, which can automatically identify and track boar sperm with a recognition rate of 96.08% and an average tracking rate of 86%. The results show that the proposed smartphone-based hand-held platform can effectively replace the traditional microscope compound computer to diagnose sperm activity. In contrast, the platform is smaller, easier to use and is not limited by the usage scenarios.

Phosphatidylserine Levels in Seminal Plasma of Iraqi Infertile Men Using Direct and Indirect Swim Up Sperm Preparation Techniques

Phosphatidylserine (PS), exposed on viable sperm, is required for fertilization as it is recognized by specific receptors located on the microvilli of the oocyte to promote sperm: egg fusion. The heads of viable and motile sperm exhibit PS. With (PS) exposure progressively increasing during sperm transit through the epididymis. PS asymmetry on the leaflets of plasma membrane is arranged in a complex way, via multiple proteins functioning in opposite directions; further, PS is also essential for many intracellular trafficking events. The aim of this study was to evaluate the phosphatidylserine PS level in seminal plasma of infertile men before and after activation using Direct Swim Up (DSU) and Indirect Swim Up (IDSU) activation method and make a comparison between them. Forty infertile men were included in this study during their attendance to the infertility clinic at High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University. Normozoospermic or mild asthenozoospermic semen samples were used to choose the cases, according to WHO manual standards (1999 -2010). The collected semen samples obtained were divided into three parts: The first part prepared was as an in vitro sperm characterization before activation while the second and third parts were after activation using direct and indirect swim up techniques respectively. The level of seminal plasma phosphatidylserine was increased after the activation techniques from the baseline level before activation. Comparison of Seminal plasma phosphatidylserine levels between direct and indirect sperm activation technique were significantly higher in direct swim up activation technique than indirect one. Phosphatidylserine (PS) in semen plasma of infertile men was significantly increased after activation using Direct Swim Up DSU preparation method.

The Potential of Theophylline and Pentoxifylline in Sperm Optimization and Its Intracytoplasmic Sperm Injection Outcomes.

Sperm motility is an essentialselection criteria by embryologists at the time of intracytoplasmic sperm injection (ICSI). One method of testing sperm viability is to induce sperm motility by increasing cyclic adenosine monophosphate (cAMP) levels by treating a semen sample with phosphodiesterase inhibitors (PDEIs), such as theophylline and pentoxifylline. It explores the implications of PDEI in medical care, reflecting on its effects in clinical settingsand recognizing potential topics for future exploration. This analysis revealed that by incorporating stimulants that activate movements, the time it took to single out sperms was markedly reduced, and consequently, the sperms were safeguarded from a prolonged period of oxidative stress. Furthermore, theophylline was found to advance sperm motility, consequently resulting in several initially immobile spermatozoa displaying rapid progressive motility. Higher fertilization rate, cleavage rate, good quality embryos (grade I), and higher biochemical and clinical pregnancy rates were found with artificial sperm activation (ASA) using pentoxifylline and theophylline. This review emphasizes the need for more research to evaluate the drug's long-term safety and investigate the effects of theophylline and pentoxifylline on postfertilization parameters, such as embryo development, implantation, and pregnancy outcomes. These areas of investigation are important for understanding the complete impact of these agents and to ensure their safe and effective implementation in clinical practice.

The Role of Soluble Adenylyl Cyclase in the Regulation of Flagellar Motility in Ascidian Sperm.

Flagellar motility in sperm is activated and regulated by factors related to the eggs at fertilization. In the ascidian Ciona intestinalis, a sulfated steroid called the SAAF (sperm activating and attracting factor) induces both sperm motility activation and chemotaxis. Cyclic AMP (cAMP) is one of the most important intracellular factors in the sperm signaling pathway. Adenylyl cyclase (AC) is the key enzyme that synthesizes cAMP at the onset of the signaling pathway in all cellular functions. We previously reported that both transmembrane AC (tmAC) and soluble AC (sAC) play important roles in sperm motility in Ciona. The tmAC plays a major role in the SAAF-induced activation of sperm motility. On the other hand, sAC is involved in the regulation of flagellar beat frequency and the Ca2+-dependent chemotactic movement of sperm. In this study, we focused on the role of sAC in the regulation of flagellar motility in Ciona sperm chemotaxis. The immunochemical analysis revealed that several isoforms of sAC protein were expressed in Ciona sperm, as reported in mammals and sea urchins. We demonstrated that sAC inhibition caused strong and transient asymmetrization during the chemotactic turn, and then sperm failed to turn toward the SAAF. In addition, real-time Ca2+ imaging in sperm flagella revealed that sAC inhibition induced an excessive and prolonged Ca2+ influx to flagella. These results indicate that sAC plays a key role in sperm chemotaxis by regulating the clearance of [Ca2+]i and by modulating Ca2+-dependent flagellar waveform conversion.

Environmentally relevant exposure to tetrabromobisphenol A induces reproductive toxicity via regulating glucose-6-phosphate 1-dehydrogenase and sperm activation in Caenorhabditis elegans

Tetrabromobisphenol A (TBBPA), a ubiquitous brominated flame-retardant environmental pollutant, has been reported to cause reproductive toxicity by chronic exposure. However, the acute reproductive risk and mechanisms of TBBPA toxicity to individuals, especially at environmentally relevant levels, remains a topic of debate. In this study, Caenorhabditis elegans was used to investigate the reproductive toxicity of acute exposure to TBBPA at environmentally relevant doses. The reproductive end points (embryonic lethality ratio and brood size), oxidative stress, sperm activation, and molecular docking were evaluated. Results showed that, after 24 h of TBBPA treatment, even at the lowest concentration (1 μg/L), the embryonic lethality ratio of C. elegans increased significantly, from 1.63 % to 3.03 %. Furthermore, TBBPA induced oxidative stress with significantly increased expression of sod-3 in C. elegans, which further raised the level of reproductive toxicity through inhibiting the activation of sperm in nematodes. In addition, molecular docking suggested TBBPA might compete for the glucose-6-phosphate-binding site of glucose-6-phosphate 1-dehydrogenase, resulting in oxidative stress generation. Accordingly, our findings indicate that even acute exposure to environmental concentrations of TBBPA may induce reproductive toxicity through reducing sperm activation in nematodes.

Оптимизация состава криопротектора для сохранения репродуктивных клеток африканского сома

The article presents a study on the selection of the optimal composition of the protective medium for cryopres-ervation of the reproductive cells of male African catfish. Glycerol, dimethyl sulfoxide (DMSO) at concentrations of 3, 5 and 10% were studied as the main cryoprotective additives. The influence of these compounds on the quality indicators of catfish sperm (the percentage and time of sperm movement after activation), the level of sperm lipid peroxidation, and the activity of the antioxidant enzyme catalase before and after the cryopreservation process was assessed. A decrease in fish breeding indicators of the quality of African catfish sperm (the proportion of living cells, motility time) was established when DMSO was added to the basic cryomedium. The greatest decrease in the percentage of sperm motility before freezing is observed when DMSO is added at a concentration of 10%. A dose-dependent effect of a decrease in sperm activity after freezing was noted with an increase in the addition of DMSO to the cryogenic medium from 3 to 10%. It has been shown that 3% glycerol in a cryogenic medium does not contribute to the preservation of cells during freezing, cryodamage caused by the formation of ice crystals leads to a significant decrease in the motility of catfish sperm. The best indicators of sperm quality were obtained when using glycerol, the most effective concentration is 5% (40% of mobile reproductive cells within 53 s). The addition of 5% glycerol to the cryomedium contributes to the greatest decrease in the level of peroxidation of sperm lipids and an increase in catalase activity, which is fully consistent with the results on sperm motility. The calculated correlation coefficients confirm the dependence of the reproductive qualities of gametes on the antioxidant status of cells. The results indicate the importance of using antioxidants as cryoprotective additives to improve the quality of African catfish sperm breeding and indicate the need for more research to evaluate the effect of adding new potentially effective and safe antioxidants as cryoprotective additives.

The role of sex hormones and ROS values in the control of andrology in male double-spurred Francolin (&lt;i&gt;Francolinus bicalcaratus&lt;/i&gt;)

Background: This study explored the relationship between andrology, seasonal sex hormone fluctuations, and ROS values in adult male Francolinus bicalcaratus while establishing baseline parameters.&#x0D; Methods: Double-spurred Francolins (n=5 per season) were randomly selected in both dry and rainy seasons from their natural habitat. They were carefully stabilized with dewormer, antibiotics, and multivitamins, then acclimatized for two weeks at the Experimental Animal House, University of Ibadan, Nigeria. The birds were weighed, sedated, and 3 to 5 ml of blood was collected from the jugular vein in lithium heparinized bottles for serum sex hormone analysis. Testes were excised, weighed, washed with a 1.15% KCL solution, and processed for complete andrology and oxidative stress assays.&#x0D; Results: Sperm count, activity, and morphological characteristics peaked during late rainy seasons, coinciding with increased mating and hatchability. This correlated with high serum testosterone and low ROS titres in the testes. Conversely, early dry seasons witnessed declines in serum testosterone, sperm parameters, and hatchability due to increased ROS titres from food scarcity. Late dry seasons saw further declines in sex hormone levels and elevated ROS titres, leading to the absence of sperm cells.&#x0D; Conclusion: This study highlights how sex hormone and ROS titres influence sperm cell viability in double-spurred Francolins, with significant seasonal variations. These factors mainly support spermatogenesis and fertility during the breeding season, characterized by food abundance and cover. Dry season sex hormone values serve as baselines, while wet season sperm count and ROS levels represent baseline data for this bird species.

Effect of L-proline on sperm quality during cryopreservation of boar semen

L-proline has been reported to be useful in semen cryopreservation. However, its use has rarely been reported in the freezing of boar semen. The objective of this study was to evaluate the effects of different concentrations of L-proline (0, 10, 30, 50, and 90 mM) on the quality of boar semen after freezing and thawing. Semen samples from boars (n = 6) were frozen using freezing extenders with added concentrations of L-proline. Total sperm motility, progressive motility, survival time at 37 °C, acrosome integrity, mitochondrial activity, DNA integrity, the content of the lipid peroxidation product malondialdehyde (MDA), total antioxidant capacity (T-AOC) and, expression levels of apoptosis protein (cleaved caspase 3 and Bax) were evaluated after thawing. The results showed that total sperm viability (73.96% vs. 63.58%) and progressive motility (56.88% vs. 47.26%) after thawing were significantly higher in the 10 mM L-proline treatment group than in the control group. The survival time at 37 °C and the total motility of sperm in the 10 mM group within one hour after thawing were significantly higher than in the control group. Acrosome integrity and mitochondrial activity of sperm in the 10 mM group were significantly higher than those in the control, 50 mM, and 90 mM groups. The DNA integrity rate in the 10 mM group was significantly higher than in the control group. The L-proline treatment did not affect sperm MDA content or T-AOC. The expression levels of apoptosis protein (cleaved caspase 3 and Bax) in the 10 mM L-proline supplemented group were lower than those in the control group. In conclusion, the freezing extender containing 10 mM L-proline improved semen quality after freezing and thawing and thus would be a useful reagent for boar semen cryopreservation.

SPIN-4/Spinster supports sperm activation in C. elegans via sphingosine-1-phosphate transport

Spermiogenesis, a sperm-activation step, is crucial for the transformation of immotile spermatids into motile sperm. Though membrane transport of ions and molecules across the sperm plasma membrane has been implicated in this process, the full repertoire of transporters involved, and their respective substrates, is unclear. Here, we report that the major facilitator superfamily transporter SPIN-4/Spinster governs efficient spermiogenesis and fertility in the hermaphrodite nematode Caenorhabditis elegans. Unlike other C. elegans Spinster paralogs, SPIN-4 is germline-expressed. Moreover, SPIN-4 expression is gamete-specific; it is strongly expressed in developing sperm, where it localizes to the plasma membrane, but it is absent from oocytes. Consistent with these expression data, we demonstrate that knocking out spin-4 impairs sperm development, leading to the formation of non-motile sperm that lack pseudopodia. Consequently, hermaphrodites homozygous for the spin-4(knu1099) knockout allele show extensive sperm wasting and reduced self-progeny. We observe similar defects when we genetically inhibit production of sphingosine-1-phosphate, a lipid molecule that stimulates cell motility when exported extracellularly by Spinster homologs in other contexts. Remarkably, extracellular supplementation with sphingosine-1-phosphate rescues sperm activation and motility in the absence of SPIN-4, suggesting that Spinster-dependent efflux of sphingosine-1-phosphate plays a key role in sperm mobilization. These findings identify a new signaling mechanism in C. elegans spermiogenesis entailing Spinster and sphingosine-1-phosphate.

Кінематичні показники та дихальна активність деконсервованих сперміїв баранів за додавання наноцитрату Mn, Zn та Cu до середовища для кріоконсервування

The aim of the work was to find out the effect of adding nanocitrate of Mn, Zn and Cu to the medium for cryopreservation of ram sperm on kinematic indicators and respiratory activity of thawed sperm. The experiment was conducted on six clinically healthy breeder rams of the Texel breed aged 2–4 years. After receiving the ejaculates of the rams, they were evaluated for the volume, concentration and motility of the sperm and were divided into control and experimental groups. Control sperm samples were diluted with lactose-yolk-tris-citrate-glycerol medium (LYTCGM). Nanocitrates of trace elements were added to the medium in experimental samples of ram sperm in the following doses: Zn and Mn — 2.5, 5.0 and 7.5 μg/l, Cu — 1.25, 2.5 and 3.75 μg/l. Diluted sperm was packaged in straws, equilibrated for 2.5 hours and frozen. After thawing of sperm, motility, morphological damage of sperm, kinematic parameters of sperm motility (CASA), oxidation and reduction activity of sperm were determined. A dose-dependent effect of Mn, Zn, and Cu nanocitrates upon their addition to LYTCGM was established. The addition of Mn and Zn nanocitrate at a dose of 5.0 μg/l to LYTCGM significantly (P&lt;0.05–0.01) increases the activity of thawed ram sperm, while the addition of Cu nanocitrate in increasing doses significantly reduces the motility of sperm in thawed ram sperm. Addition of Mn and Zn nanocitrate in an optimal dose of 5.0 μg/l to LYTCGM significantly (P&lt;0.05–0.01) reduces the number of spermatozoa degenerated and with damaged acrosomes, and with the addition of Cu nanocitrate in increasing doses, morphological disorders of germ cell significantly increase cells The addition of Mn and Zn nanocitrate at a dose of 5.0 μg/l to LYTCGM significantly (P&lt;0.01–0.001) increases the kinematic parameters of thawed ram sperm, and the addition of Cu nanocitrate in increasing doses significantly reduces the indicators of germ cell motility. The addition of Mn and Zn nanocitrates to the medium for cryopreservation of ram sperm increases the oxidation and inhibits the reduction activity of thawed sperm. Addition of Cu nanocitrate to LYTCGM in increasing doses reduces the oxidation and increases the reduction activity of thawed ram sperm.

D-Limonene Alleviates Oxidative Stress Injury of the Testis Induced by Arsenic in Rat.

Long-term exposure to arsenic can lead to testicular damage and lower sperm quality in males, which is mediated by increased arsenic-induced oxidative stress and other damage mechanisms. D-Limonene, which is rich in oranges, lemons, oranges, grapes and other natural fruits, can relieve doxorubicin (DOX)-induced kidney injury and CCL4-induced cardiac toxicity by inhibiting oxidative stress and inflammatory response. The antioxidant and anti-inflammatory properties of D-limonene motivate us to further explore whether it can reduce arsenic-induced testicular injury. To verify this scientific hypothesis, testicular pathology, testicular oxidative stress levels and sperm motility were determined after intervention with D-limonene in rats chronically exposed to arsenic. As expected, long-term arsenic exposure caused testicular tissue structure disturbances, increased levels of oxidative stress, and decreased sperm activation, all of which were significantly inhibited due to treatment with D-limonene. In conclusion, our data reveal a previously unproven beneficial effect of D-limonene, namely that D-limonene can inhibit arsenic-induced testicular injury, and also provide theoretical and experimental basis for the application of D-limonene in the treatment of arsenic-induced testicular injury.

Kaempferol Enhances Sperm Post-Thaw Survival by Its Cryoprotective and Antioxidant Behavior

This study examined the effects of three selected doses of kaempferol (KAE; 12.5, 25 or 50 μM) on bovine sperm motility and oxidative profile directly related to cold storage. We also elucidated the effect of KAE on the expression profiles of heat shock proteins (HSPs) 70 and 90 as well as the pro-apoptotic BCL2-associated X (BAX) protein and the anti-apoptotic B-cell lymphoma 2 (Bcl-2) protein. Frozen samples supplemented with KAE were compared with a native control (fresh spermatozoa) and a cryopreserved control, frozen in the absence of KAE. Our results demonstrate that the administration of all KAE doses led to a higher degree of sperm motility (p &lt; 0.05) when compared with the cryopreserved control. The highest levels of protection of sperm DNA (p &lt; 0.05), lipids (p &lt; 0.05) and proteins (p &lt; 0.05) were detected in samples exposed to 25 μM KAE when compared with samples frozen without KAE. Administration of 25 μM KAE led to a significant increase in HSP70 and HSP90 (p &lt; 0.05) when compared with the unsupplemented frozen control. No significant differences were observed in the expression patterns of BAX; however, a significant up-regulation of Bcl-2 protein was observed in the frozen samples enriched with 25 μM KAE when compared with the cryopreserved control (p &lt; 0.05). In summary, we may consider KAE as an effective agent in stabilizing the sperm membranes by preventing reactive oxygen species (ROS) overproduction in the mitochondria and subsequent oxidative damage to molecules critical for a proper sperm architecture and function. These protective properties of KAE may lead to higher post-thaw sperm activity and viability.

Effect of Some Psychotic Drug Quetiapine on Reproductive System Activity in Male Rats

Aim of study: To investigate the effects of antipsychotic medications utilized to treat schizophrenic patients and its effects on the reproductive system. Introduction: Antipsychotics, anticonvulsants, and other all psychotropic drugs have negative effects on the quality of sperm and sexual activity. These negative side effects differ amongst males and its less severe for some drugs, enabling some degree of control over their effects. Sperm can suffer oxidative damage from spending too much time in the male reproductive system. One of the antipsychotics most frequently administered to treat schizophrenia in adults is quetiapine. In this work, the effects of repeated rats’ healthy production of sperm in response to therapeutic doses of quetiapine was studied. Rats were also used to assess quetiapine’s effects on hormonal balance and oxidative state. Methodology: The experiment employed male wistar rats weighing 300 to 350 g at 10 to 12 weeks of age. The experiment employed male wistar rats weighing 300 to 350 g at 10 to 12 weeks of age. Rats received oral dosages of quetiapine for 30 days. At this time’s conclusion, the body’s weights and the organs were analyzed, additionally to sperm concentration, motility, shape, and degree of sperm damage. The levels of testosterone, LH, and other male hormones related to reproduction were measured in the serum. Malondialdehyde and glutathione levels were measured to assess the oxidative state of testicular tissues. Results: The results of this investigation demonstrated that in rats receiving quetiapine, aberrant sperm morphology increased while relative epididymis weights and sperm concentration dropped. Rats receiving quetiapine experienced a drop in serum LH and testosterone levels. Rats receiving quetiapine also had lower amounts of malondialdehyde, which was assessed. Conclusion: Quetiapine therapy this negative effect may be attributed to lower sperm quality, changed hormone levels, and enhanced oxidative stress.