Acrosome Integrity Research Articles

Evaluating the Cryoprotective Effects of Butylated Hydroxytoluene on Semen Quality Parameters of Phasianus colchicus

Cryopreservation is critical for the preservation of genetic resources and the propagation of endangered species. The choice of proper cryoprotectants significantly affects the success of cryopreservation. The current study investigated the impact of butylated-hydroxytoluene (BHT) on sperm microscopic assays in Ring-necked Pheasants at various post-processing phases, including post-dilution, post-cooling, post-equilibration, and post-thawing. Semen samples were collected and cryopreserved at various times with BHT (0.0 mM (control), 0.5 mM, 1.0 mM, 1.5 mM, and 2.0 mM. Acrosomal integrity (AI), viability, plasma membrane integrity (PMI), and sperm motility were all assessed as significant markers of sperm quality. It was discovered that using a 1.0 mM concentration of BHT had a significant (p > 0.05) influence on each stage of cryopreservation, resulting in improvements in sperm motility, PMI, viability, and AI when analyzed in both control and other groups. These findings emphasize BHT's cryoprotective qualities of BHT in protecting the quality and integrity of Ring-necked Pheasant sperm during cryopreservation. Further research is needed to understand the mechanisms by which BHT exerts its protective effects and to maximize its use in assisted reproductive treatments. The use of BHT as a cryoprotectant has the potential to aid in the conservation of endangered bird species, such as the Ring-necked Pheasant.

Impact of Trolox Supplementation on the Cryopreservation of Honamli Buck Semen.

Cryopreservation of buck semen is essential in animal breeding but often damages sperm viability and integrity. The Honamli breed, a hardy Turkish goat, can benefit from improved freezing techniques using antioxidants such as Trolox (T). This study explores the effects of varying T concentrations on Honamli buck semen, assessing parameters such as motility, viability, and membrane integrity to enhance post-thaw quality. Findings support T's potential to improve semen extender formulations for preserving Honamli genetics.This study aims to freeze Honamli buck semen with T and to evaluate in vitro spermatological parameters. Three Honamli bucks, aged 2-3 years, were used in the study. Semen was collected from the bucks and mixed after removing seminal plasma. The mixed semen was diluted with a tris egg yolk extender containing three different concentrations of T (0.25 mM, 0.5 mM, and 1 mM) and control (0 mM). The diluted semen was equilibrated for 2 hours at +4 degrees and subjected to cryopreservation in liquid nitrogen vapor (-120°C for 12 minutes) and frozen. After thawing (37°C water bath for 30 seconds), the groups were evaluated at flow cytometric analysis for viability (SYBR/propidium iodide [PI]), plasma membrane acrosome integrity (FITC-PNA/PI), and mitochondrial membrane potential (JC-1), plasma membrane integrity (hypo-osmotic swelling test), microscopic evaluations for motility and morphological integrity (abnormal spermatozoa rate). The 0.5 T and 0.25 T groups showed significant improvements in motility compared with the control group (p < 0.05). The control group had the lowest plasma membrane integrity (p < 0.05). The highest morphological integrity was observed in the T groups compared with the control group (p < 0.05). In conclusion, supplementing T in buck semen extenders benefits spermatological parameters; particularly, 0.25 and 0.5 mM T could be used in Tris semen extenders during the cryopreservation process.

Influence of antifreeze protein III on canine sperm cryopreservation.

Influence of antifreeze protein III on canine sperm cryopreservation.

Beneficial effects of EGCG on boar sperm quality during liquid storage at 4°C are mediated by DRD2 receptor.

Beneficial effects of EGCG on boar sperm quality during liquid storage at 4°C are mediated by DRD2 receptor.

Identification of key genes and variants associated with boar sperm freezability using whole genome resequencing.

Identification of key genes and variants associated with boar sperm freezability using whole genome resequencing.

Spirulina polysaccharides improve postthaw sperm quality in bulls by inhibiting the activation of pathways related to protein kinase A.

Sperm cryopreservation is widely used in assisted reproductive technology (ART) and livestock breeding. Although sperm cryopreservation accelerates breeding, the quality of cryopreserved sperm tends to be decreased. Improving the quality of frozen sperm is a hot topic, and spirulina polysaccharide, known for its immunomodulatory and antioxidant properties, is considered a promising natural extract for extensive studies. In this study, a pectic polysaccharide was extracted from spirulina (PSP), and its effects on postthaw bovine sperm viability were evaluated. Phosphoproteomic analysis based on TMT labelling and LC-MS/MS was employed. The results revealed that 10mg/L PSP had significant protective effects on postthaw sperm viability, plasma membrane integrity, acrosomal integrity, and mitochondrial membrane integrity. Moreover, PSP increased the antioxidant capacity by activating antioxidant enzymes such as SOD, CAT, and GSH-PX and reduced apoptosis, ROS release and MDA levels. In addition, PSP resulted in decreased phosphorylation levels of proteins related to the acrosome, flagellum, metabolism, energy acquisition, and apoptosis. This protective effect of PSP on frozen sperm was achieved by inhibiting the activation of protein kinase A(PKA) protein-related pathways.

Incorporating olive (Olea europaea L) fruit extracts in a tris-based extender improves buffalo semen cryotolerance by reducing oxidative stress.

This work aimed to evaluate whether supplementing the freezing extender with olive fruit extract (OFE) would improve the antioxidant defense of buffalo sperm, resulting in improved post-thaw semen quality. Ejaculates (two per 16 Murrah buffalo bulls) were split into four aliquots that were diluted in an extender supplemented with different doses of OFE (0, D50, D100, and D150, based on µM concentrations of hydroxytyrosol, the most represented polyphenol) and frozen according to standard procedures. At thawing, sperm motility, kinetics, viability, acrosome integrity, and membrane functionality were evaluated. Based on the dose-response results, biological antioxidant potential (BAP) and reactive oxygen metabolites (ROMs) were assessed after thawing in D50 and control groups. The pre-freezing supplementation of the extender with D50 OFE showed higher (P < 0.05) total and progressive sperm motility, as well as straight-line velocity compared to the control. Treatment with D50 OFE of buffalo semen also improved (P < 0.01) post-thaw sperm viability, membrane functionality, and acrosome integrity compared to the control. The enrichment of the extender with D50 OFE increased (P < 0.01) the post-thaw BAP and reduced (P < 0.05) the ROMs levels. The highest concentration tested (D150 OFE) negatively affected (P < 0.05) total and progressive motility, and the percentage of sperm with functional membranes and intact acrosomes, compared to the control. In conclusion, low doses of OFE added to the extender significantly improved post-thawing buffalo semen quality by protecting the spermatozoa from cryopreservation-induced oxidative stress. Further studies should investigate its effectiveness on in vivo and in vitro fertility, for potential commercial applications.

Protective effect of supplementation with water soluble β-carotene and α-tocopherol in boar sperm cooling-freezing extender, but not in the thawing extender.

Protective effect of supplementation with water soluble β-carotene and α-tocopherol in boar sperm cooling-freezing extender, but not in the thawing extender.

Impact of particle size of nanoliposomes on the biological response of frozen-thawed sperm in Holstein bulls.

Impact of particle size of nanoliposomes on the biological response of frozen-thawed sperm in Holstein bulls.

Polydopamine-based nano-protectant for prolonged boar semen preservation by eliminating ROS and regulating protein phosphorylation via D2DR-mediated cAMP/PKA signaling pathway

IntroductionPreservation of porcine semen is essential for artificial insemination and genetic improvement in pig breeding programs. However, the overproduction of reactive oxygen species (ROS) and lower levels of protein phosphorylation emerge as two challenges during semen preservation. Inspired by the innate ligand-receptor binding biofunction of dopamine, herein, a dual-task nano-protectant that combines ROS-scavenging and protein phosphorylation-regulating properties via incorporating the natural antioxidant epigallocatechin gallate (EGCG) into polydopamine nanoparticles (EGCG@PDA NPs) was proposed to enhance the quality of pig semen during storage at 4 ℃. The results suggested that EGCG@PDA NPs significantly maintained sperm motility, acrosome integrity and mitochondrial membrane potential, extending semen storage time from 3 days to 10 days. Furthermore, EGCG@PDA NPs effectively scavenged excess ROS and inhibited ROS-mediated sperm apoptosis through the extracellular regulated protein kinases (ERK) signaling pathway. Intriguingly, EGCG@PDA NPs could degrade into ultrasmall particles (< 10 nm) in the semen or H2O2 systems. These particles could target and activate the dopamine D2 receptor (D2DR) on membrane surface of sperm midpiece, thereby enhancing protein phosphorylation via the downstream cyclic adenosine monophosphate/protein kinase A (cAMP/PKA) signaling pathway, ultimately improving sperm motility parameters. This study presents a novel nano-strategy to boost the quality of pig semen, offering significant implications for the pig industry.Graphical

Intracytoplasmic Motile Sperm Selection in Fresh and Frozen Semen and its Correlation with Seminal Attributes of Kankrej Bull

Total 8 mature healthy Kankrej bulls (7 replicates) were selected for evaluation of seminal attributes at fresh and frozen stage of cryopreservation. Seminal attributes evaluated included sperm viability, acrosome integrity, HOST-EN test, intracytoplasmic motile sperm selection (IMSI) and sperm mucus penetration test (SMPT). Based on the nuclear abnormalities, sperms were classified in Grade I, II, III and IV. Per cent sperm viability, acrosome integrity, HOST- reacted sperm and Grade I sperm were found significantly lower in frozen semen as compared to fresh semen. Grade IV sperm as assessed by IMSI tool was found significantly higher in frozen semen as compared to fresh semen. Correlation study revealed significant and positive correlations of sperm viability with acrosome integrity and HOST, while significant negative correlation with grade IV sperm. Sperm viability and acrosome integrity were positively correlated with SMPT. Key words: Cryopreservation, IMSI, Kankrej bull, Semen, SMPT

Effect of Centrifuged Chicken Egg Yolk on the Cryopreservation of Boar Semen.

Egg yolk, commonly employed as a cryoprotectant in semen cryopreservation, contains large particle matter that can diminish semen quality post thaw and complicate its quality assessment. For this reason, we designed a centrifugal treatment of chicken egg yolk to evaluate its effect on the cryopreservation of porcine semen. The control group (CG) was prepared with a dilution of chicken egg yolk by conventional mixing treatment, and the experimental group (EG) used a dilution of centrifugally treated chicken egg yolk for the ultra-low-temperature cryopreservation of porcine semen. The freezing process was carried out by conventional freezing methods. The spermatozoa were subsequently assessed for various parameters, including motility, acrosome integrity rate, plasma membrane integrity rate, antioxidant indexes, apoptosis rate, and the expression of apoptosis-related genes. The results showed that, post freeze-thawing, the motility, viability, VSL, and VCL of the spermatozoa in the EG were significantly higher than those observed in the CG (p < 0.05). Additionally, the acrosome integrity and plasma membrane integrity of the spermatozoa in the EG were significantly enhanced compared to the CG (p < 0.05). Furthermore, the EG exhibited significantly lower MDA content and sperm apoptosis rate (p < 0.05), while demonstrating significantly higher T-AOC and CAT levels (p < 0.05) relative to the CG. In comparison to the CG, the EG exhibited a significant reduction in the gene expression of TNF-a and Bax in the spermatozoa (p < 0.05), whereas the expression levels of CAT and Bcl-2 were significantly elevated (p < 0.05). In conclusion, the dilution solution formulated through the centrifugal processing of chicken egg yolk demonstrated efficacy in enhancing the quality of porcine spermatozoa following cryopreservation and subsequent thawing.

The Sarcoplasmic/Endoplasmic reticulum Ca2+-ATPase (SERCA) is present in pig sperm and modulates their physiology over liquid preservation

Liquid storage is the primary preservation method in the swine breeding industry because of its advantages over cryopreservation. Calcium (Ca2+), a key regulator of cell physiology, plays a crucial role during liquid preservation. Sarcoplasmic/Endoplasmic Reticulum Ca2+ ATPases (SERCA) belong to a family of P-type ATPases that regulate Ca2+ homeostasis within cells and have been previously described to play a function in the sperm of various mammalian species. Herein, we hypothesized that SERCA2 is present in pig sperm and is involved in the resilience of this cell to liquid preservation at 17 °C. For this purpose, sperm were incubated with different concentrations of thapsigargin (Thg; 0, 5, 25, and 50 µM) and stored at 17 °C for ten days. The presence and localization of SERCA2 were evaluated using immunoblotting and immunofluorescence, respectively. On days 0, 4, and 10, sperm motility was assessed using a computer-assisted sperm analysis (CASA) system, and sperm viability, membrane lipid disorder, acrosome integrity, mitochondrial membrane potential (MMP), and intracellular levels of Ca2+, superoxides and total reactive oxygen species (ROS) were evaluated by flow cytometry. We localized SERCA2 in the acrosome and midpiece of pig sperm. Furthermore, inhibition of SERCA with Thg resulted in reduced sperm viability and membrane stability, and increased MMP, and Ca2+ and ROS levels. In conclusion, the activity of SERCA prevents the accumulation of intracellular Ca2+ in sperm, which is detrimental to sperm quality and function during liquid storage at 17 °C. We thus suggest that the function of SERCA is crucial for the preservation of pig semen.

MitoQ Preserves Epigenetic Modifications and Quality Parameters of Rooster Sperm During Cryopreservation Process.

Cryopreservation in rooster semen is a helpful procedure to spread qualified semen samples for reproductive goals. Nevertheless, some post-thawed qualified semen samples showed a considerably poor fertility rate that might be related to epigenetic modifications during the cryopreservation process. This study aims to investigate the effect of the cryopreservation process in the presence of MitoQ as a mitochondrial-targeted antioxidant on epigenetic changes and other quality parameters (motility, morphology, mitochondrial activity, acrosome integrity, lipid peroxidation, DNA fragmentation, apoptosis status, and ROS concentration) of rooster sperm. The collected semen samples were divided into four groups of fresh samples and three groups that were supplemented by MitoQ 0, 10, and 100 nM and cryopreserved. The cryopreservation process reduced (p ≤ 0.05) DNA methylation, H3K9 acetylation, H3K4 methylation, motility parameters, membrane integrity, mitochondrial activity, acrosome integrity, viability, and increased (p ≤ 0.05) lipid peroxidation, DNA fragmentation, ROS concentration, and apoptotic-like changes compared to the fresh semen group. However, in frozen sperm groups, MitoQ 10 and 100 nM resulted in significant improvements (p ≤ 0.05) in the epigenetic modifications and other mentioned quality parameters compared to the control group (MitoQ 0). Generally, although the cryopreservation process reduced semen quality, using MitoQ could be useful in cryopreserved rooster semen quality.

Mitochondria-targeted antioxidant MitoQ improves the quality of low temperature-preserved yak semen via alleviating oxidative stress.

Mitochondria-targeted antioxidant MitoQ improves the quality of low temperature-preserved yak semen via alleviating oxidative stress.

Bupropion, the atypical antidepressant used in smoke cessation: an in vitro study on its effects on human sperm function.

Bupropion, the atypical antidepressant used in smoke cessation: an in vitro study on its effects on human sperm function.

Effect of Vitamin K Additive to Tris-Egg Yolk Extender on Iraqi Buffalo Bull Semen Characteristics

Abstract Buffalo sperm are sensitive to peroxidation of lipids because buffalo spermatozoa contain highly polyunsaturated free fatty acid (PUFA) which reduces the bull fertility following cryopreservation compared to other species. The current study was carried out to determine the effect of vitamin K supplementation in tris extender on Iraqi buffalo semen following treatment, cooling, and cryopreservation. Four Iraqi buffalo bulls were used in the current study. Samples were collected from all bulls accumulated and extended in tris extender with vitamin K at 0 (control), 0.03, 0.06, 0.09 ml/100 extender before cryopreservation. Individual motility, live sperm, total abnormal sperm, acrosome integrity, and integrity of the sperm plasm membrane were estimated in this study. The results showed the low values of abnormal sperms and high value of intact membrane, intact acrosome, and live sperm were recorded with 0.03 ml vitamin K. In conclusion, 0.03 ml vitamin K is highlighted in improving the Iraqi buffalo semen characteristics and lowering the freezing–thawing damages. The current outcome is warranted to estimate the vitamin K effect on the parameters of antioxidant.

Molecular mechanisms of libido influencing semen quality in geese through the hypothalamic-pituitary-testicular-external genitalia axis.

Molecular mechanisms of libido influencing semen quality in geese through the hypothalamic-pituitary-testicular-external genitalia axis.

The dual nature of micronutrients on fertility: too much of a good thing?

The dual nature of micronutrients on fertility: too much of a good thing?

Age-Related Dynamics in the Conventional, Non-Conventional, and Bacteriological Characteristics of Fresh and Liquid-Stored Porcine Semen.

This study strove to investigate the effect of boar age on conventional and non-conventional quality traits of fresh and liquid-stored semen. Sixty boars were allocated into 3 groups: 8-12 months (young); 24-36 months (adult); and 48-60 months (senior). Ejaculates were divided into two parts; the first one was assessed in native state while the second one was extended in the Androstar Plus extender containing gentamycin, stored at 5 °C and evaluated following 72 h. Young animals presented with a significantly lower sperm motility (p < 0.01), membrane and acrosome integrity (p < 0.0001), and mitochondrial activity (p < 0.0001) against adult boars. Significantly higher levels of free radicals and tumor necrosis factor alpha (p < 0.001), interleukin 1 and 6 (p < 0.0001) were found in young boars in comparison to adult boars. The assessment of liquid-stored semen revealed a significantly lower sperm motility, membrane, and acrosome integrity (p < 0.0001) in young boars when compared to adult boars. Moreover, Clostridium difficile, Escherichia coli, Pseudomonas aeruginosa, and Rothia nasimurium remained in liquid-stored semen obtained from young boars, while Corynebacterium sp. and Escherichia coli continued to be identified in samples collected from adult boars. In conclusion, age contributes to the overall quality of fresh as well as liquid-stored boar semen.