Exogenous Oleic Acid and Palmitic Acid Improve Boar Sperm Motility via Enhancing Mitochondrial Β-Oxidation for ATP Generation

It takes several hours for mammalian sperm to migrate from the ejaculation or insemination site to the fertilization site in the female reproductive tract in which glucose, amino acids, and fatty acids are regarded as the primary substrates for ATP generation. The present study was designed to investigate whether oleic acid and palmitic acid were beneficial to boar sperm in vitro; and if yes, to elucidate the mechanism that regulates sperm motility. Therefore, the levels of oleic acid and palmitic acid, motility, membrane integrity, acrosome integrity, and apoptosis of sperm were evaluated. Moreover, the enzymes involved in mitochondrial β-oxidation (CPT1: carnitine palmitoyltransferase 1; ACADVL: long-chain acyl-coenzyme A dehydrogenase) were detected with immunofluorescence and Western blotting. Consequently, the ATP content and the activities of CPT1, ACADVL, malate dehydrogenase (MDH), and succinate dehydrogenase (SDH) were also measured. We observed that CPT1 and ACADVL were expressed in boar sperm and localized in the midpiece. The levels of oleic acid and palmitic acid were decreased during storage at 17 °C. The addition of oleic acid and palmitic acid significantly increased sperm motility, progressive motility, straight-line velocity (VSL), membrane integrity, and acrosome integrity with a simultaneous decrease in sperm apoptosis after seven days during storage. When sperm were incubated with oleic acid and palmitic acid at 37 °C for 3 h, the activities of CPT1 and ACADVL, the ATP level, the mitochondrial membrane potential, the activities of MDH and SDH, as well as sperm motility patterns were significantly increased compared to the control (p < 0.05). Moreover, the addition of etomoxir to the diluted medium in the presence of either oleic acid or palmitic acid and the positive effects of oleic acid and palmitic acid were counteracted. Together, these data suggest that boar sperm might utilize oleic acid and palmitic acid as energy substrates for ATP production via β-oxidation. The addition of these acids could improve sperm quality.

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Effect of L-carnitine on sperm quality during liquid storage of boar semen

Asian-Australasian Journal of Animal Sciences ◽

10.5713/ajas.19.0455 ◽

2020 ◽

Vol 33 (11) ◽

pp. 1763-1769

Author(s):

Kang Yang ◽

Na Wang ◽

Hai-Tao Guo ◽

Jing-Ran Wang ◽

Huan-Huan Sun ◽

...

Keyword(s):

Sperm Motility ◽

Sperm Quality ◽

Membrane Integrity ◽

Quality Parameters ◽

Atp Content ◽

Developmental Potential ◽

Positive Effects ◽

Liquid Storage ◽

Boar Sperm

Objective: This study was conducted to investigate the effect of L-carnitine on the pig semen characteristics during storage.Methods: Spermatozoa samples were examined for spermatozoa quality and then randomly divided into 5 groups: 0 (control), 12.5, 25, 50, and 100 mM L-carnitine. Sperm motility, plasma membrane integrity and antioxidant parameters (total reactive oxygen species, total antioxidant capacity, and malondialdehyde) were evaluated after 0, 3, 5, and 10 day cooledstorage at 17°C. Moreover, ATP content, mitochondria activity as well as sperm-binding and in vitro fertilizing ability of preserved boar sperm were also investigated.Results: Supplementation with 50 mM L-carnitine could effectively maintain boar sperm quality parameters such as sperm motility and membrane integrity. Besides, we found that L-carnitine had positive effects on boar sperm quality mainly through improving antioxidant capacities and enhancing ATP content and mitochondria activity. Interestingly, by assessing the effect of L-carnitine on sperm fertility and developmental potential, we discovered that the extender containing L-carnitine could improve sperm quality and increase the number of sperms bounding to zona pellucida, without improving in vitro fertility and development potential.Conclusion: These findings suggested that the proper addition of L-carnitine to the semen extender improved boar sperm quality during liquid storage at 17°C.

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Effects of Taurine on Sperm Quality during Room Temperature Storage in Hu Sheep

Animals ◽

10.3390/ani11092725 ◽

2021 ◽

Vol 11 (9) ◽

pp. 2725

Author(s):

Liuming Zhang ◽

Yanhu Wang ◽

Tariq Sohail ◽

Yan Kang ◽

Haoyuan Niu ◽

...

Keyword(s):

Room Temperature ◽

Sperm Quality ◽

Membrane Integrity ◽

Quality Parameters ◽

Control Group ◽

Negative Effects ◽

Positive Effects ◽

Antioxidant Parameters ◽

Acrosome Integrity ◽

Hu Sheep

The present study aimed to investigate whether the presence of Tau protected Hu sheep sperm from ROS stress during storage at room temperature. The semen was diluted with extender (Tris-based) at room temperature, supplemented with different concentrations of Tau (0, 10, 20, 40, 80, or 100 mM), and stored at 15 °C. Sperm quality parameters (sperm progressive motility, kinetic parameters, plasma membrane integrity rate, acrosome integrity rate, and MMP) and antioxidant parameters (ROS, MDA, SOD, CAT, and T-AOC) were evaluated during the preservation of semen. The addition of Tau, especially at a concentration of 20 mM, exerted positive effects on sperm quality parameters and antioxidant parameters compared to the sperm without Tau treatment (control group). The addition of Tau, especially at a concentration of 100 mM, exerted negative effects on sperm quality parameters and antioxidant parameters compared to the control group. Interestingly, the results indicated that the sperm acrosome integrity rate did not change during storage time. In conclusion, the addition of Tau to sperm preserved at room temperature can enhance the antioxidant ability of sperm, reduce the LPO on the 5th day, and improve the quality of semen preserved at room temperature. These results implied that Tau had potential to enhance Hu sheep sperm reproductive performance.

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Resveratrol protects boar sperm in vitro via its antioxidant capacity

Zygote ◽

10.1017/s0967199420000271 ◽

2020 ◽

Vol 28 (5) ◽

pp. 417-424

Author(s):

Linlin Sun ◽

Xiaoteng Fan ◽

Yao Zeng ◽

Liqian Wang ◽

Zhendong Zhu ◽

...

Keyword(s):

Antioxidant Capacity ◽

Sperm Motility ◽

Mitochondrial Membrane ◽

Membrane Integrity ◽

B Cell Lymphoma ◽

Membrane Potentials ◽

Cooling Process ◽

Boar Sperm ◽

Fast Cooling ◽

Acrosome Integrity

SummaryThe objective of the present study was to elucidate whether resveratrol could facilitate the survival of boar sperm during liquid preservation and fast cooling processes. Boar semen were diluted with Modena extender containing different concentrations of resveratrol. Sperm motility was evaluated by visual estimation. Membrane integrity, acrosome integrity and mitochondrial membrane potentials were measured by SYBR-14/PI, FITC-PNA and JC-1 staining, respectively. Moreover, the levels of reactive oxygen species (ROS), malonaldehyde (MDA) and total antioxidant capacity (T-AOC) were measured using commercial assay kits. B-cell lymphoma protein-2 (BCL2) content was determined by western blotting. During liquid preservation at 17oC, the addition of 50 μM resveratrol to the Modena extender significantly improved sperm motility, membrane integrity, acrosome integrity, and sperm mitochondrial membrane potentials. Similar results were also observed in the 150 μM resveratrol group during the fast cooling process. Furthermore, addition of resveratrol led to a decrease of ROS and MDA, and an increase in the content of T-AOC and BCL2. These observations suggest that addition of resveratrol to Modena extender protects boar sperm against oxidative stress. The optimal concentrations of resveratrol are 50 μM and 150 μM during liquid preservation and fast cooling process, respectively.

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Effects of the platelet-activating factor (PAF) on selected quality parameters of cryopreserved bull semen (AI) with reduced sperm motility

Polish Journal of Veterinary Sciences ◽

10.1515/pjvs-2016-0019 ◽

2016 ◽

Vol 19 (1) ◽

pp. 147-158 ◽

Cited By ~ 2

Author(s):

M. Lecewicz ◽

W. Kordan ◽

A. Majewska ◽

S. Kamiński ◽

A. Dziekońska ◽

...

Keyword(s):

Plasma Membrane ◽

Sperm Motility ◽

Sperm Quality ◽

Membrane Integrity ◽

Platelet Activating Factor ◽

Quality Parameters ◽

Sperm Viability ◽

Plasma Membrane Integrity ◽

Positive Effects ◽

Bull Semen

Abstract The aim of the study was to determine the effects of platelet-activating factor (PAF) on selected quality parameters of cryopreserved bull semen with reduced sperm motility used for artificial insemination. The aim of experiment 1 was to identify the optimal concentration of the phospholipid able to preserve sperm viability. Cryopreserved semen was treated with different PAF concentrations: 1×10-5M, 1×10-6M, 1×10-7M, 1×10-8M and 1×10-9M. The experiment demonstrated that PAF at concentration 1×10-9M increased most the sperm viability parameters (motility parameters, plasma membrane integrity and mitochondrial function) after 120 min of incubation of thawed semen at 37°C. Cryopreserved bull semen with reduced sperm motility (below 70%) was supplemented with PAF in a concentration of 1×10-9M. A statistically significant increase in sperm motility, percentage of linear motile spermatozoa and VSL value was observed after 120 min incubation of sperm with 1×10-9M PAF. Sperm supplementation with PAF also had positive effects on plasma membrane integrity and percentage of spermatozoa with preserved mitochondrial transmembrane potential, but the differences were not statistically significant. The results indicated positive effects of PAF supplementation at a concentration of 1×10-9M on the selected sperm quality parameters in cryopreserved bull semen with reduced motility.

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Butaphosphan and Cyanocobalamin Supplementation in Semen Extender on Chilled Boar Sperm Quality and Life Span

Frontiers in Veterinary Science ◽

10.3389/fvets.2020.592162 ◽

2020 ◽

Vol 7 ◽

Author(s):

J. Suwimonteerabutr ◽

S. Chumsri ◽

P. Tummaruk ◽

Morakot Nuntapaitoon

Keyword(s):

Plasma Membrane ◽

Life Span ◽

Sperm Quality ◽

Membrane Integrity ◽

Control Group ◽

Plasma Membrane Integrity ◽

Progressive Motility ◽

Boar Sperm ◽

Acrosome Integrity ◽

Semen Extender

The objective of the present study was to determine the effect of butaphosphan and cyanocobalamin supplementation in semen extender on chilled boar sperm quality and life span. A total of 35 ejaculates of boar semen were included. The semen was diluted with Beltsville thawing solution extender supplemented with different concentrations of butaphosphan and cyanocobalamin [0 (control), 0.1, 0.2, 0.3, 0.4, and 0.5%] in the diluted semen. The semen samples were evaluated using a computer-assisted sperm analysis system to determine sperm motility and sperm kinetic parameters (i.e., the curvilinear velocity, VCL; straight line velocity, VSL; average path velocity, VAP; linearity, LIN; straightness, STR; amplitude of lateral head, ALH; wobble, WOB; and beat cross frequency, BCF). Additionally, sperm viability, acrosome integrity, mitochondrial activity, and plasma membrane integrity were evaluated after 4 (day 0), 72 (day 3), 120 (day 5), and 168 (day 7) h of storage using SYBR-14–ethidium homodimer-1 (EthD-1), EthD-1, JC-1, and the short hypo-osmotic swelling test, respectively. The analyses were carried out by using the general linear mixed model (MIXED) procedure of SAS. The statistical models for each data set included group, day after storage, and interaction between group and day after storage. The boar was included as a random effect. On day 0 after storage, progressive motility, VCL, VSL, VAP, and plasma membrane integrity of boar sperm in 0.3% of butaphosphan and cyanocobalamin supplementation were greater than those in the 0.4 and 0.5% groups (P < 0.05). On day 3 after storage, total motility and progressive motility, VCL, VSL, VAP, LIN, WOB, BCF, and plasma membrane integrity in 0.3% of butaphosphan and cyanocobalamin supplementation were significantly greater than those in the control group (P < 0.05). The total motility and progressive motility, VAP, and WOB in 0.3% of butaphosphan and cyanocobalamin supplementation were greater than those in the control group on day 5 after storage (P < 0.05). No effects of butaphosphan and cyanocobalamin supplementation on acrosome integrity and mitochondria activity were found on days 3, 5, and 7 after storage. However, the motility and progressive motility and the values for all sperm kinetic parameters except ALH in 0.3% of butaphosphan and cyanocobalamin supplementation were greater than those in the control group on day 7 after storage (P < 0.05). In conclusion, 0.3% of butaphosphan and cyanocobalamin supplementation in semen extender improved sperm motility, sperm activity, morphology, and life span in chilled boar sperm.

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A comparative study of two methods to determine acrosome integrity of frozen-thawed boar sperm

Veterinarska stanica ◽

10.46419/vs.52.6.4 ◽

2021 ◽

Vol 52 (6) ◽

Author(s):

Janyaporn Rungruangsak ◽

Junpen Suwimonteerabutr ◽

Kakanang Buranaamnuay ◽

Arun Chankrachang ◽

Padet Tummaruk

Keyword(s):

Sperm Quality ◽

Membrane Integrity ◽

Blue Staining ◽

Computer Assisted ◽

Sperm Viability ◽

Coomassie Blue Staining ◽

Coomassie Blue ◽

Boar Sperm ◽

Acrosome Integrity ◽

Sperm Acrosome

Coomassie blue staining has been reported as an effective and inexpensive method for evaluating the acrosome integrity of spermatozoa, though to date its use to evaluate cryopreserved boar sperm has not been reported. Moreover, there is no information concerning the agreement between Coomassie blue staining and fluorescein isothiocyanate conjugated peanut agglutinin and ethidium homodimer (FITC-PNA/EthD-1) methods for assessing sperm acrosome integrity for any species. The current study was performed to determine the efficacy and agreement between Coomassie blue and FITC-PNA/EthD-1 staining methods for evaluating the acrosome integrity of frozen-thawed boar sperm. A total of 25 semen samples were cryopreserved using lactose-egg yolk-based extender and loaded into 0.5 PVC-French straws. Sperm motility and motion characteristics were determined using a computer-assisted sperm analysis system. Sperm viability and plasma membrane integrity were evaluated using the SYBR-14/EthD-1 and hypo-osmotic swelling test, respectively. Acrosome integrity of frozen-thawed boar sperm was evaluated using both FITC-PNA/EthD-1 and Coomassie blue staining to assess the association between sperm acrosome integrity and agreement between these two methods. The average percent acrosome integrity of frozen-thawed boar sperm as determined by FITC-PNA/ EthD-1 and Coomassie blue staining was 48.8 ± 12.6% and 52.6 ± 13.6%, respectively (P>0.05). Interestingly, Coomassie blue staining found a correlation between sperm viability and acrosome integrity (r=0.609, P=0.002), while FITC-PNA/EthD-1 staining did not (P>0.05). However, the acrosome integrity of frozen-thawed boar sperm evaluated by FITC-PNA/ EthD-1 and Coomassie blue staining was significantly correlated (r=0.448, P=0.025, n=25). The Bland-Altman plot determined that this agreement was acceptable. In conclusion, the acrosome integrity of the frozen-thawed boar sperm assessed via Coomassie blue staining was significantly correlated with that obtained via the FITC-PNA/EthD-1 staining method, and the two methods showed good agreement. Moreover, the significant association between the acrosome integrity of frozen-thawed boar sperm determined by Coomassie blue staining with other sperm quality parameters indicates that this is an effective method for assessing the acrosome integrity of frozen-thawed sperm in pigs.

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Effect of docosahexanoic acid on quality of frozen–thawed bull semen in BioXcell extender

Reproduction Fertility and Development ◽

10.1071/rd15089 ◽

2017 ◽

Vol 29 (3) ◽

pp. 490 ◽

Cited By ~ 3

Author(s):

Asmatullah Kaka ◽

Wahid Haron ◽

Rosnina Yusoff ◽

Nurhusien Yimer ◽

A. M. Khumran ◽

...

Keyword(s):

Lipid Peroxidation ◽

Liquid Nitrogen ◽

Sperm Motility ◽

Membrane Integrity ◽

Optimum Level ◽

Docosahexanoic Acid ◽

Normal Morphology ◽

Bull Semen ◽

Acrosome Integrity

This study was conducted to investigate the effect of docosahexanoic acid (DHA) supplementation in BioXcell extender on the quality of frozen–thawed bull semen. Twenty-four ejaculates were collected from three bulls (eight from each bull). Ejaculates with motility ≥70% and normal morphology ≥80% were extended into BioXcell extender to which 0 (control), 3, 5, 10 or 15 ng mL–1 DHA was added. The supplemented semen samples were incubated at 37°C for 15 min for DHA uptake by spermatozoa. Later, samples were cooled for 2 h at 5°C and packaged into 0.25-mL straws, frozen in liquid nitrogen for 24 h and subsequently thawed for evaluation. Results are presented as percentages ± s.e.m. Supplementation with DHA at 3 ng mL–1 significantly improved sperm functional parameters including sperm motility, normal morphology, viability, acrosome integrity and membrane integrity when compared with other supplemented groups and the control. Lipid peroxidation increased as the incorporation of DHA supplementation increased. In conclusion, 3 ng mL–1 concentration of DHA resulted in superior quality of frozen–thawed bull spermatozoa and is suggested as the optimum level of DHA to be added into BioXcell extender.

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Antioxidant Effect of a Polyphenol-Rich Murtilla (Ugni molinae Turcz.) Extract and Its Effect on the Regulation of Metabolism in Refrigerated Boar Sperm

Oxidative Medicine and Cellular Longevity ◽

10.1155/2019/2917513 ◽

2019 ◽

Vol 2019 ◽

pp. 1-15 ◽

Cited By ~ 3

Author(s):

Ignacio Jofré ◽

Magdalena Cuevas ◽

Leticia Signori de Castro ◽

João Diego de Agostini Losano ◽

Mariana Andrade Torres ◽

...

Keyword(s):

Lipid Peroxidation ◽

Sperm Motility ◽

Sperm Quality ◽

Membrane Damage ◽

Antioxidant Effect ◽

Boar Sperm ◽

Fertilization Capacity ◽

Ugni Molinae

The production of reactive oxygen species (ROS) in boar spermatozoa increases in refrigeration; this can have an impact on sperm quality and fertilization capacity. We evaluated the effect of polyphenol-rich aqueous extract of murtilla (Ugni molinae Turcz) on boar sperm stored at 17°C in order to reduce oxidative stress and improve sperm quality in the long term. Five experiments were performed: first, characterization of the polyphenol content from five genotypes of murtilla; second, determination of the genotype with the best antioxidant effect (MT-Ex); third, the antioxidant capacity on O2- and lipid peroxidation; fourth, the influence of MT-Ex on motility, calcium movement, cAMP, and metabolic parameters; and fifth, analysis of long-term refrigeration. The average phenolic content was 344 ppm; gallic acid, catechin, quercetin, myricetin, and kaempferol were detected. All extracts evaluated presented a concentration-dependent antioxidant effect. MT-Ex reduces intracellular O2-/peroxides but low lipid peroxidation. MT-Ex in nonstimulated ROS conditions reduces sperm motility, mitochondrial membrane potential, cAMP, and ATP, but the succinate dehydrogenase activity remained normal; also, we observed a reduction in calcium movement in in vitro sperm capacitation. The long-term analyses showed that MT-Ex improved sperm motility decay and reduced membrane damage and ROS at 168 h. Based on this study, we propose MT-Ex as a supplement in semen extenders.

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Screening Mold Colonies by Using Two Toxicity Assays Revealed Indoor Strains of Aspergillus calidoustus Producing Ophiobolins G and K

Toxins ◽

10.3390/toxins11120683 ◽

2019 ◽

Vol 11 (12) ◽

pp. 683 ◽

Cited By ~ 2

Author(s):

Marja Johanna Salo ◽

Tamás Marik ◽

Ottó Bencsik ◽

Raimo Mikkola ◽

László Kredics ◽

...

Keyword(s):

Mass Spectrometry ◽

Cell Proliferation ◽

Sperm Motility ◽

High Performance ◽

Biological Activities ◽

Membrane Integrity ◽

Opportunistic Pathogen ◽

Mass Spectrometry Data ◽

Toxic Substances ◽

Boar Sperm

The occurrence and toxin production of the opportunistic pathogen Aspergillus calidoustus in Finnish buildings is not well documented in the literature. We tracked and identified four A. calidoustus colonies cultivated from indoor settled dusts and revealed the biological activities of crude biomass extracts. The toxic substances were identified as 6-epi-ophiobolin K, ophiobolin K, and ophiobolin G by high-performance liquid chromatography–mass spectrometry (HPLC-MS) based on chromatographic and mass spectrometry data (MS and MS/MS) on the crude extract of A. calidoustus strain MH34. A total of 29 fungal colonies collected from settled dust in an office room reported for indoor-air-related illnesses were screened for toxins that inhibited boar sperm motility in the BSMI (boar sperm motility inhibiting) assay and cell proliferation in the ICP (inhibition of cell proliferation) assays with PK-15 cells. Out of the 27 colonies tested as toxic, 12 colonies exhibiting conidiophores representative of the genera Chaetomium, Penicillium, and Paecilomyces were excluded from the study, while 13 colonies exhibited Aspergillus-like conidiophores. Biomass suspensions of these colonies were divided into two categories: Category 1 colonies (n = 4), toxic in the BSMI assay and the ICP assays, emitted blue fluorescence and grew at 37 °C; Category 2 colonies (n = 9), only toxic in the ICP assay, emitted orange fluorescence and exhibited limited or no growth at 37 °C. Colonies in Category 1 were pure-cultured, and the strains were named as MH4, MH21, MH34, MH36. Strain MH34 was identified as A. calidoustus by the internal transcribed spacer (ITS) sequences. Ethanol-soluble dry substances extracted from the biomass of the pure cultures exhibited a toxicological profile in the BSMI assay, SMID (sperm membrane integrity damage) assay, and ICP assay similar to that exhibited by pure ophiobolin A. Overall, the viable conidia of A. calidoustus in indoor settled dusts deserve attention when potentially hazardous mold species are monitored.

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The effects of chronic administration of ghrelin on rat sperm quality and membrane integrity

Animal Biology ◽

10.1163/157075609x437682 ◽

2009 ◽

Vol 59 (2) ◽

pp. 159-168 ◽

Cited By ~ 11

Author(s):

Arash Kheradmand ◽

Majid Taati ◽

Homayoon Babaei

Keyword(s):

Plasma Membrane ◽

Treatment Group ◽

Sperm Motility ◽

Sperm Quality ◽

Sperm Concentration ◽

Membrane Integrity ◽

Chronic Administration ◽

Control Group ◽

Plasma Membrane Integrity ◽

Significant Difference

AbstractAlthough ghrelin acts as a modulator of feeding behavior and energy metabolism in the central nervous system, recent studies have implicated the peripheral actions of ghrelin in reproductive tissues. Here, we investigated the effects of chronic administration of ghrelin on the motility, plasma membrane integrity and concentration of rat spermatozoa. 45-d male Wistar rats were scheduled for the study and were divided into control and treatment groups. In the treatment group, 1 nmol of ghrelin was administered as sc injection for 10 consecutive days or vehicle (physiological saline) to the control rats. Sperm collection was achieved by killing of the rats on days 15, 25 and 50 after first injection. Total sperm motility and forward progressive movement did not exhibit significant difference during the experiment, although, there was a tendency for greater motion rate on d 15 and 25 in the treated rats compared to the control group. Plasma membrane integrity (HOS-reacted spermatozoa) was significantly higher in the treated animals, especially on day 15 as well as day 25, because of possible antioxidant properties of ghrelin. This value was statistically higher on day 15 than that of day 25 (P <0.05). Likewise, there was a significant correlation between the FPM (P <0.0001, r = 0.79) and TSM (P <0.01, r = 0.52) with the HOS test percentage in the treatment group. It was not observed statistically difference in the sperm concentration between groups during all of the experimental days. In conclusion, chronic administration of ghrelin (similar to induced by energy deficiency such as fasting) increased the integrity of sperm membrane, however, the sperm motility and concentration did not display any alterations.

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