scholarly journals Effects of various physical stress factors on mitochondrial function and reactive oxygen species in rat spermatozoa

2013 ◽  
Vol 25 (7) ◽  
pp. 1051 ◽  
Author(s):  
Suhee Kim ◽  
Cansu Agca ◽  
Yuksel Agca
Keyword(s):  
Reactive Oxygen Species ◽  
Membrane Integrity ◽  
Reactive Oxygen ◽  
Careful Consideration ◽  
Stress Factors ◽  
Oxygen Species ◽  
Percoll Gradient ◽  
Functional Parameters ◽  
Positive Correlations ◽  
Gradient Separation

The aim of the present study was to evaluate the effects of various physical interventions on the function of epididymal rat spermatozoa and determine whether there are correlations among these functional parameters. Epididymal rat spermatozoa were subjected to various mechanical (pipetting, centrifugation and Percoll gradient separation) and anisotonic conditions, and sperm motility, plasma membrane integrity (PMI), mitochondrial membrane potential (MMP) and intracellular reactive oxygen species (ROS) were evaluated. Repeated pipetting caused a loss in motility, PMI and MMP (P < 0.05). Minimal centrifugation force (200g) had no effect on motility, PMI and MMP, whereas an increase in the centrifugation force to 400g or 600g decreased sperm function (P < 0.005). Percoll gradient separation increased total motility, PMI and MMP (P < 0.05). However, the spermatozoa that were subjected to mechanical interventions showed high susceptibility to a ROS stimulant (P < 0.005). Anisotonic conditions decreased motility, PMI and MMP, and hypotonic conditions in particular increased basal ROS (P < 0.05). In correlation tests, there were strong positive correlations among total motility, PMI and MMP, whereas ROS showed no or negatively weak correlations with the other parameters. In conclusion, the physical interventions may act as important variables, affecting functional parameters of epididymal rat spermatozoa. Therefore, careful consideration and proper protocols for handling of rat spermatozoa and osmotic conditions are required to achieve reliable results and minimise damage.

69 REACTIVE OXYGEN SPECIES EVALUATION OF DONKEY FROZEN SEMEN ADDED TO HOMOLOGOUS SEMINAL PLASMA ON POST-THAW

2015 ◽  
Vol 27 (1) ◽  
pp. 127
Author(s):  
P. V. L. Oliveira ◽  
J. V. Oliveira ◽  
C. Ramires Neto ◽  
Y. F. R. Sancler-Silva ◽  
C. P. Freitas-Dell'aqua ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Kinetic Parameters ◽  
Seminal Plasma ◽  
Membrane Integrity ◽  
Reactive Oxygen ◽  
Computer Assisted ◽  
Oxygen Species ◽  
Inflammation Response ◽  
Frozen Semen ◽  
Uterine Inflammation

For many years the pregnancy rate of donkey frozen semen presented lower results in donkey jennies; however, a recent study showed an increase in pregnancy rates using frozen semen added to seminal plasma on post-thaw. A hypothesis for this result is the higher uterine inflammation response after breeding when using seminal plasma. The same studies demonstrated higher uterine inflammation in the presence of higher reactive oxygen species concentration. The aim of the present study was to evaluate the content of reactive oxygen species in donkey frozen semen added to homologous seminal plasma on post-thaw. Five ejaculates from each 3 donkeys were used. Semen was diluted (1 : 1) with a skim milk-based extender (Botu-SemenTM, Botupharma, Brazil). The semen was frozen with Botu-CryoTM extender (Botupharma, Brazil) in an isothermal box in straws containing 100 × 106 of total sperm. The samples were thawed at 46°C for 20 s. After this, the straws of each donkey were divided in 2 group: control group (CG), in which the semen was incubated at 37°C for 5 min, and plasma seminal group (PG), in which the semen was incubated at 37°C for 5 min with 70% of homologous seminal plasma. Sperm kinetic parameters were evaluated by computer-assisted semen analysis, and the plasma membrane integrity (propidium iodide and fluorescein isothiocyanate -PSA) and reactive oxygen species (5–6-carboxi-2,7-diclorodihidrofluoresceindiacetate) were evaluated by flow cytometer. Comparison of sperm parameters was performed by t-test. Total motility (%, CG = 75.4 ± 8.2a v. PG = 57.5 ± 16.4b), progressive motility (%, CG = 42.0 ± 8.7a v. PG = 33.3 ± 13.2b), progressive angular velocity (μm/s, CG = 95.8 ± 10.8a v. PG = 88.9 ± 10.9b), and percentage of rapid sperm (%, CG = 58.4 ± 12.5a v. PG = 41.0 ± 17.3b) were higher in CG compare with PG. No difference (P < 0.05) was observed in membrane integrity (%, CG = 20.7 ± 7.4 v. PG = 20.6 ± 7.8); however, reactive oxygen species (%, CG = 12.3 ± 10.6a v. PG = 81.8 ± 32.5b) were higher in PG. The results of this study showed that the addition of homologous seminal plasma on post-thaw decreases the sperm kinetic parameters and viability of donkey frozen semen but increases reactive oxygen species, and this may cause higher uterine inflammation response in donkey jennies and increase their fertility.

Reactive Oxygen Species and RBC Membrane Integrity in Peroxyredoxin II Deficient Mice.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 54-54
Author(s):  
Amrita Bhagat ◽  
Renee Emerson ◽  
Kitty DeJong ◽  
Frans A. Kuypers
Keyword(s):  
Reactive Oxygen Species ◽  
Antioxidant Properties ◽  
Flow Cytometric Analysis ◽  
Membrane Integrity ◽  
Reactive Oxygen ◽  
Extinction Time ◽  
Oxygen Species ◽  
Rbc Membrane ◽  
Small Proteins ◽  
Phospholipid Scrambling

Abstract Red blood cells (RBCs) contain a complex set of enzymes and non-enzymatic scavengers as defense against reactive oxygen species (ROS). Peroxiredoxin II (PrxII), a member of a family of small proteins with strong antioxidant properties, is highly abundant in RBCs. PrxII is likely to play an essential role in ROS protection, as RBCs generate high levels of ROS due to their role in oxygen transport and the presence of redox active hemoglobin. Phosphatidylserine (PS) asymmetry in RBCs is maintained by the active transport of PS from outer to inner monolayer by the oxidant sensitive aminophospholipid translocase or flipase. PS exposure is found in RBCs where phospholipid scrambling is activated and the flipase is inhibited. In PrxII−/− mice, PrxII is absent from the RBC. These mice are anemic (Hct 41% vs. 46% in wild type mice (WT)), with increased reticulocyte count (4.7% vs. 2.0 % in WT), and a morphologically diverse RBC population. RBC indices after isovolumetric sphering showed a similar MCH (14.3 vs. 14.2), slightly increased MCV (49.2 fl vs. 45.5 fl) and slightly decreased MCHC (30.1 vs. 32.1) in PrxII−/− mice as compared to WT. In flow cytometric analysis, two distinct populations of RBC are found with either slight or significantly increased autofluorescence in the fluorescein channel (excitation 488 nm, emission 515 nm), indicative of oxidant damage. These two populations of low (LF) and high (HF) fluorescent cells comprise 70–80% and 20–30% of the total RBC population respectively. RBC from PrxII−/− and WT mice were biotinylated using EZ-Link Sulfo-NHS-Biotin (Pierce) allowing turnover studies of the LF and HF population. At set time points, the number of biotinylated cells was determined in small blood samples by flow cytometry using fluorescently labeled streptavidin. The data were mathematically fitted to 100–100*[1−(1/T)*t]exp(−kt), where t is the time point, T is the extinction time, and k the exponential rate of RBC removal. The data in the WT showed a linear removal rate (k=0), and a T of 40 days (R2=0.99). In PrxII−/−, an overall faster disappearance of biotinylated cells was noted, and the number of surviving (biotinylated) cells in the population followed an exponential pattern, consistent with random removal (k=0.08, R2= 0.98). At 20 days, 50% of biotinylated RBC were present in WT, but only 18% were found in PrxII−/− mice. In the non-biotinylated RBC, HF cells started to appear at day 13, indicating that autofluorescence is acquired in time. Using the fluorescent ROS membrane probe C11-BODIPY, our data indicate a higher level of ROS in the HF population. The HF population exhibited a lower flipase activity and increased phospholipid scrambling, as measured by labeling with annexin V. Together, our data indicate the importance of PrxII in the maintenance of RBC membrane integrity and suggest that oxidant induced PS exposure is in part responsible for shortened RBC survival in these mice. These findings indicate a role for oxidation in the exposure of PS on the RBC surface, which may clarify mechanisms in oxidant induced membrane alterations in hemoglobinopathies.

Specific inhibition of mitochondrial oxidative stress suppresses inflammation and improves cardiac function in a rat pneumonia-related sepsis model

2012 ◽  
Vol 302 (9) ◽  
pp. H1847-H1859 ◽  
Author(s):  
Qun S. Zang ◽  
Hesham Sadek ◽  
David L. Maass ◽  
Bobbie Martinez ◽  
Lisha Ma ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Vitamin E ◽  
Antioxidant Capacity ◽  
Membrane Integrity ◽  
Reactive Oxygen ◽  
Myocardial Inflammation ◽  
Oxygen Species ◽  
Mitochondrial Reactive Oxygen Species ◽  
Sprague Dawley ◽  
Sepsis Model

Using a mitochondria-targeted vitamin E (Mito-Vit-E) in a rat pneumonia-related sepsis model, we examined the role of mitochondrial reactive oxygen species in sepsis-mediated myocardial inflammation and subsequent cardiac contractile dysfunction. Sepsis was produced in adult male Sprague-Dawley rats via intratracheal injection of S. pneumonia (4 × 106colony formation units per rat). A single dose of Mito-Vit-E, vitamin E, or control vehicle, at 21.5 μmol/kg, was administered 30 min postinoculation. Blood was collected, and heart tissue was harvested at various time points. Mito-Vit-E in vivo distribution was confirmed by mass spectrometry. In cardiac mitochondria, Mito-Vit-E improved total antioxidant capacity and suppressed H2O2generation, whereas vitamin E offered little effect. In cytosol, both antioxidants decreased H2O2levels, but only vitamin E strengthened antioxidant capacity. Mito-Vit-E protected mitochondrial structure and function in the heart during sepsis, demonstrated by reduction in lipid and protein oxidation, preservation of mitochondrial membrane integrity, and recovery of respiratory function. While both Mito-Vit-E and vitamin E suppressed sepsis-induced peripheral and myocardial production of proinflammatory cytokines (tumor necrosis factor-α, interleukin-1β, and interleukin-6), Mito-Vit-E exhibited significantly higher efficacy ( P < 0.05). Stronger anti-inflammatory action of Mito-Vit-E was further shown by its near-complete inhibition of sepsis-induced myeloperoxidase accumulation in myocardium, suggesting its effect on neutrophil infiltration. Echocardiography analysis indicated that Mito-Vit-E ameliorated cardiac contractility of sepsis animals, shown by improved fractional shortening and ejection fraction. Together, our data suggest that targeted scavenging of mitochondrial reactive oxygen species protects mitochondrial function, attenuates tissue-level inflammation, and improves whole organ activities in the heart during sepsis.

Effect of adding heterologous versus homologous bovine seminal plasma prior to cryopreservation on bull sperm quality after thawing

Zygote ◽  
2018 ◽  
Vol 26 (5) ◽  
pp. 388-394
Author(s):  
Thanapol Nongbua ◽  
Essraa M Al-Essawe ◽  
Anders Edman ◽  
Anders Johannisson ◽  
Jane M Morrell
Keyword(s):  
Reactive Oxygen Species ◽  
Seminal Plasma ◽  
Deleterious Effect ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Reactive Oxygen ◽  
Low Fertility ◽  
Computer Assisted ◽  
High Fertility ◽  
Oxygen Species

SummaryThe aim of this study was to investigate the effect of adding homologous or heterologous bovine seminal plasma (SP) to SP-free sperm samples before freezing on sperm quality after thawing. Ejaculates from bulls of known fertility were used as a source of SP. The SP was removed from further aliquots of the same ejaculates by colloid centrifugation to create SP-free sperm samples; the resuspended sperm pellets were treated with homologous or heterologous SP from high or low fertility bulls at 0%, 1% or 5% before freezing. After thawing, sperm quality was evaluated by computer-assisted sperm analysis and flow cytometry for membrane integrity, reactive oxygen species, chromatin structure, mitochondrial membrane potential and protein tyrosine phosphorylation. Data were analysed using Proc MIXED, SAS®. Post-hoc comparisons were adjusted for multiplicity using Tukey’s method. The addition of SP resulted in significant differences in sperm quality, namely velocity class A, Velocity Straight Line (VSL), Velocity Average Path (VAP), Velocity Curved Line (VCL), Amplitude of Lateral Head Displacement (ALH), Hyperactive (HYP), reactive oxygen species (ROS) production and % DNA fragmentation index (DFI) (P<0.05 for each). Although adding 5% homologous SP from high fertility bulls was beneficial to sperm kinematics, 5% heterologous SP from high fertility bulls had a deleterious effect on chromatin integrity and on sperm velocity. In conclusion, adding SP may have either a beneficial effect or a deleterious effect depending on the individuals involved. It might be feasible to use this method to improve sperm quality in some circumstances.

scholarly journals Improved Post-Thaw Quality of Canine Semen after Treatment with Exosomes from Conditioned Medium of Adipose-Derived Mesenchymal Stem Cells

Animals ◽  
2019 ◽  
Vol 9 (11) ◽  
pp. 865 ◽  
Author(s):  
Ahmad Qamar ◽  
Xun Fang ◽  
Min Kim ◽  
Jongki Cho
Keyword(s):  
Reactive Oxygen Species ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Group 2 ◽  
And Function

Freezing decreases sperm quality, ultimately affecting fertilizing ability. The repair of freeze-damaged sperm is considered crucial for improving post-thaw viability and fertility. We investigated the effects of exosomes derived from canine adipose-derived mesenchymal stem cells on dog sperm structure and function during cryopreservation. The pooled ejaculate was diluted with buffer, without (Control), or with exosomal proteins (25, 50, or 100 µg/mL). Using fresh semen, the determined optimal exosomal protein concentration was 50 µg/mL (Group 2) which was used in further experiments. Post-thaw sperm treated with exosomes were superior to control (p < 0.05) in terms of motility (56.8 ± 0.3% vs. 47.2 ± 0.3%), live sperm percentage (55.9 ± 0.4% vs. 45.4 ± 0.4%), membrane integrity (55.6 ± 0.5% vs. 47.8 ± 0.3%), and acrosome integrity (60.4 ± 1.1% vs. 48.6 ± 0.4%). Moreover, expression of genes related to the repair of the plasma membrane (ANX 1, FN 1, and DYSF), and chromatin material (H3, and HMGB 1) was statistically higher in exosome-treated sperm than control, but the expression of the mitochondrial reactive oxygen species modulator 1 gene was significantly higher in control. Therefore, exosomal treatment may improve the quality of post-thaw dog semen through initiating damaged sperm repair and decreasing reactive oxygen species production.

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