scholarly journals Single Layer Centrifugation with Androcoll-P Can Be Scaled-Up to Process Larger Volumes of Boar Semen

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Marjet van Wienen ◽  
Anders Johannisson ◽  
Margareta Wallgren ◽  
Joyce Parlevliet ◽  
Jane M. Morrell
Keyword(s):  
Reactive Oxygen Species ◽  
Sperm Quality ◽  
Scale Up ◽  
Membrane Integrity ◽  
Single Layer ◽  
Ros Production ◽  
Oxygen Species ◽  
Normal Morphology ◽  
Preliminary Step ◽  
Boar Semen

The objective of this study was to scale-up the procedure for Single Layer Centrifugation (SLC) through AndrocollTM-P, as a preliminary step towords processing the whole ejaculate. The first experiment compared Single Layer Centrifugation using 4.5 mL and 15 mL extended ejaculate (SLC-4.5 and SLC-15, resp.), assessing sperm quality by objective motility analysis, morphology, viability, and the production of reactive oxygen species (ROS). In the second experiment, SLC-4.5 was compared to Single Layer Centrifugation with 25 mL extended ejaculate (SLC-25) using motility analysis and morphology. In both experiments, normal morphology and linear motility were significantly higher in the SLC-selected samples than in the uncentrifuged controls (P<.001), whereas total motility and membrane integrity were unchanged. Although ROS production was higher in the SLC-selected samples than in the controls (P<.01), this might have been due to the presence of antioxidants in seminal plasma in the latter. In conclusion, there was no difference in sperm quality between SLC-4.5 and SLC-15 samples, or between SLC-4.5 and SLC-25 samples, indicating that the SLC method can be scaled-up successfully.

Effect of Single Layer Centrifugation on reactive oxygen species and sperm mitochondrial membrane potential in cooled stallion semen

2017 ◽  
Vol 29 (5) ◽  
pp. 1039 ◽  
Author(s):  
J. M. Morrell ◽  
A. Lagerqvist ◽  
P. Humblot ◽  
A. Johannisson
Keyword(s):  
Reactive Oxygen Species ◽  
Hydrogen Peroxide ◽  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Reactive Oxygen ◽  
Single Layer ◽  
Ros Production ◽  
Oxygen Species ◽  
Species Production

Additional means are needed for evaluating the quality of stallion spermatozoa in semen doses for AI. Mitochondrial membrane potential (ΔΨm) has been linked to fertility in some species, but is rarely used in the evaluation of cooled stallion semen; metabolic activity may be associated with reactive oxygen species production (ROS). In the present study, ΔΨm and ROS production were measured in doses of cooled stallion semen. The effect of colloid centrifugation on these parameters was also investigated. In this case, colloid centrifugation involves centrifuging a sperm sample through a silane-coated silica colloid formulation to retrieve the most robust spermatozoa. High and low ΔΨm in cooled stallion semen varied between stallions and between ejaculates, but was not affected by single-layer centrifugation (SLC). The SLC-selected spermatozoa produced significantly less hydrogen peroxide than controls (P < 0.001), which could explain the increased longevity and retention of fertilising capacity seen in previous studies. For SLC samples, ΔΨm was positively associated with viable spermatozoa that were not producing reactive oxygen species (r = 0.49; P < 0.001) and negatively associated with ROS production (for superoxide: r = –0.4, P < 0.01; for hydrogen peroxide: r = –0.39, P < 0.05). There was no clear association between ΔΨm and ROS production in control samples.

323 POSSIBLE BENEFICIAL EFFECT OF REACTIVE OXYGEN SPECIES ON BOAR SPERM FUNCTION

2010 ◽  
Vol 22 (1) ◽  
pp. 317
Author(s):  
C. H. C. Viana ◽  
M. Nichi ◽  
R. O. C. Silva ◽  
E. G. A. Perez ◽  
R. N. Garcia ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Hydroxyl Radical ◽  
Superoxide Anion ◽  
Present Experiment ◽  
Membrane Integrity ◽  
Physiological Role ◽  
Sperm Function ◽  
Oxygen Species ◽  
Boar Sperm ◽  
Boar Semen

Boar spermatozoa are sensitive to the attack of reactive oxygen species (ROS) due to the high content of unsaturated fatty acids in its plasma membrane and the relative low antioxidant capacity of boar seminal plasma. The benefical effect of antioxidant supplementation to the semen of boars is still a matter of debate, with different results in the literature, probably due to the empirical methods used to determine the more adequate antioxidant, and to the physiological role or the ROS. The aim of the present experiment was to study which ROS is the most deleterious to the boar sperm. Sperm samples from 4 boars were collected by digital manipulation. Sperm samples were then incubated (1 h, 37°C) with 4 ROS inducer mechanisms: xanthine/xanthine oxidase (produces superoxide anion), hydrogen peroxide (4 mM), ascorbate/ferrous sulfate (4 mM; produced hydroxyl radical), and malondialdehyde (lipid peroxidation product). Samples were analyzed for the 3-3′ diaminobenzidine stain, as an index of mitochondrial activity; the eosin/nigrosin staining, as an index of membrane integrity; the simple stain (fast green/Bengal rose), as an index of acrosome integrity; and the measurement of thiobarbituric acid reactive substances (TBARS), as an index of lipid peroxidation. Results showed that the boar sperm is relatively resistant to all ROS studied in the present experiment. Only for the percentage of sperm showing intact membrane, a difference was found between incubation with superoxide anion and hydroxyl radical (Table 1). The TBARS production was higher in samples incubated with hydroxyl radical, indicating that these samples were more susceptible to the attack of this ROS. Surprisingly, a positive correlation was found between TBARS and membrane integrity (r = 0.51, P = 0.04), indicating that samples more susceptible to the oxidative stress showed higher membrane integrity. A hypothesis to explain such conflicting results would be the physiological role of the ROS on several spermatic events, especially in this particular species. It is well known that boar semen, in order to achieve ideal motility, requires aerobic conditions. Further studies are necessary to study the effect of each ROS on each physiological or pathological event on boar sperm function. Table 1.Effect of different ROS in boar semen The authors thank Nutricell for the media used in the present experiment.

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.

scholarly journals Single Layer Centrifugation Can Be Scaled-Up Further to Process up to 150 mL Semen

2011 ◽  
Vol 2011 ◽  
pp. 1-6 ◽  
Author(s):  
J. M. Morrell ◽  
M. van Wienen ◽  
M. Wallgren
Keyword(s):  
Sperm Quality ◽  
Scale Up ◽  
Single Layer ◽  
Original Technique ◽  
Glass Tubes ◽  
Boar Semen

Single-Layer centrifugation has been used to improve the quality of sperm samples in several species. However, where stallion or boar semen is to be used for AI, larger volumes of semen have to be processed than for other species, thus limiting the effectiveness of the original technique. The objective of the present study was to scale up the SLC method for both stallion and boar semen. Stallion semen could be processed in 100 mL glass tubes without a loss of sperm quality, and similarly, boar semen could be processed in 200 mL and 500 mL tubes without losing sperm quality. The results of these preliminary studies are encouraging, and larger trials are underway to evaluate using these methods in the field.

13 REACTIVE OXYGEN SPECIES IN STORED STALLION SEMEN

2013 ◽  
Vol 25 (1) ◽  
pp. 153
Author(s):  
J. M. Morrell ◽  
A. Lundgren ◽  
P. Humblot ◽  
A. Johannisson
Keyword(s):  
Reactive Oxygen Species ◽  
Hydrogen Peroxide ◽  
Flow Cytometry ◽  
Sperm Motility ◽  
Chromatin Structure ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Pregnancy Rates ◽  
Oxygen Species ◽  
Progressive Motility

The quality of cooled semen doses for AI varies considerably between stallions. One of the factors affecting sperm quality may be the content of reactive oxygen species (ROS), which are known to affect fertility in some species. The objective of this study was to measure the ROS content of cooled stored stallion semen doses for AI as part of an evaluation of sperm quality and to correlate it with pregnancy rates. Ejaculates (3 per stallion) were collected from 14 stallions at a commercial stud farm and were extended in INRA 96 (IMV Technologies) as standard cooled semen doses for AI. After transporting the semen doses overnight to the laboratory at the Swedish University of Agricultural Sciences (Uppsala, Sweden) in an insulated container with a cold pack, aliquots were evaluated for sperm quality and ROS content by staining with hydroethidine and dihydrodichlorofluorescein diacetate and measuring fluorescence by flow cytometry (Guthrie and Welch 2006 J. Anim. Sci. 84, 2089–2100). Seven sperm sub-populations were quantified: superoxide positive or negative, living (S+L and S–L, respectively); superoxide positive, dead (S+D); and hydrogen peroxide positive or negative, living or dead (P+L, P–L, P+D, and P–D, respectively). Sperm motility was measured by computer-assisted sperm motility analysis (Sperm Vision, Minitube), membrane integrity by flow cytometry, and chromatin structure using the sperm chromatin structure assay (all assays described in Morrell et al. 2011 Theriogenology 76, 1424–1432). Pregnancy rates following AI with cooled semen doses from the same stallions were available later in the year. The effect of stallion was tested by ANOVA. Correlations were calculated between the proportions of sperm stained for S or P and other sperm quality parameters and also with pregnancy rates (SAS version 9.1, SAS Institute Inc., Cary, NC, USA). The effect of stallion was significant on all variables measured (P < 0.01). There were no significant correlations between percentages of S+L or P+L spermatozoa and progressive motility, membrane integrity, or DNA fragmentation index (%DFI). There was a trend for the proportion of P-L spermatozoa to be correlated with progressive motility (r = 0.51; P < 0.07) whereas the proportions of S+D and P–D were negatively correlated with progressive motility (r = –0.66 and –0.61; P < 0.02). The proportion of S+D and chromatin damage were negatively correlated (r = –0.54; P < 0.05). There was a trend for the proportions of S+D and P+D to be negatively related to the overall pregnancy rate (r = –0.52; P < 0.07, and r = –0.58; P < 0.05, respectively). The proportions of superoxide and hydrogen peroxide-containing live spermatozoa were not correlated to stallion fertility or other parameters of semen quality in stored semen samples. In contrast, a significant negative relationship was found between the proportion of dead spermatozoa producing superoxide radicals and progressive motility, and between dead spermatozoa producing superoxide radicals and chromatin damage. However, many factors contribute to sperm quality and fertility, with many interactions between them. More work is needed to unravel these different effects and determine which factors can be used to predict stallion fertility.

scholarly journals Reactive Oxygen Species in Host Plant Are Required for an Early Defense Response against Attack of Stagonospora nodorum Berk. Necrotrophic Effectors SnTox

Plants ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1586
Author(s):  
Svetlana Veselova ◽  
Tatyana Nuzhnaya ◽  
Guzel Burkhanova ◽  
Sergey Rumyantsev ◽  
Igor Maksimov
Keyword(s):  
Reactive Oxygen Species ◽  
Early Stage ◽  
Reactive Oxygen ◽  
Triticum Aestivum L ◽  
Redox Status ◽  
Stagonospora Nodorum ◽  
Ros Generation ◽  
Ros Production ◽  
Oxygen Species ◽  
Necrotrophic Effectors

Reactive oxygen species (ROS) play a central role in plant immune responses. The most important virulence factors of the Stagonospora nodorum Berk. are multiple fungal necrotrophic effectors (NEs) (SnTox) that affect the redox-status and cause necrosis and/or chlorosis in wheat lines possessing dominant susceptibility genes (Snn). However, the effect of NEs on ROS generation at the early stages of infection has not been studied. We studied the early stage of infection of various wheat genotypes with S nodorum isolates -Sn4VD, SnB, and Sn9MN, carrying a different set of NE genes. Our results indicate that all three NEs of SnToxA, SnTox1, SnTox3 significantly contributed to cause disease, and the virulence of the isolates depended on their differential expression in plants (Triticum aestivum L.). The Tsn1–SnToxA, Snn1–SnTox1and Snn3–SnTox3 interactions played an important role in inhibition ROS production at the initial stage of infection. The Snn3–SnTox3 inhibited ROS production in wheat by affecting NADPH-oxidases, peroxidases, superoxide dismutase and catalase. The Tsn1–SnToxA inhibited ROS production in wheat by affecting peroxidases and catalase. The Snn1–SnTox1 inhibited the production of ROS in wheat by mainly affecting a peroxidase. Collectively, these results show that the inverse gene-for gene interactions between effector of pathogen and product of host sensitivity gene suppress the host’s own PAMP-triggered immunity pathway, resulting in NE-triggered susceptibility (NETS). These results are fundamentally changing our understanding of the development of this economical important wheat disease.

scholarly journals Production and scavenging of reactive oxygen species both affect reproductive success in male and female Drosophila melanogaster

2021 ◽  
Author(s):  
Biz R. Turnell ◽  
Luisa Kumpitsch ◽  
Klaus Reinhardt
Keyword(s):  
Reactive Oxygen Species ◽  
Drosophila Melanogaster ◽  
Reactive Oxygen ◽  
Cellular Aging ◽  
Ros Production ◽  
Oxygen Species ◽  
Wild Type ◽  
Ros Scavenging ◽  
Respiratory Pathway ◽  
Male And Female

AbstractSperm aging is accelerated by the buildup of reactive oxygen species (ROS), which cause oxidative damage to various cellular components. Aging can be slowed by limiting the production of mitochondrial ROS and by increasing the production of antioxidants, both of which can be generated in the sperm cell itself or in the surrounding somatic tissues of the male and female reproductive tracts. However, few studies have compared the separate contributions of ROS production and ROS scavenging to sperm aging, or to cellular aging in general. We measured reproductive fitness in two lines of Drosophila melanogaster genetically engineered to (1) produce fewer ROS via expression of alternative oxidase (AOX), an alternative respiratory pathway; or (2) scavenge fewer ROS due to a loss-of-function mutation in the antioxidant gene dj-1β. Wild-type females mated to AOX males had increased fecundity and longer fertility durations, consistent with slower aging in AOX sperm. Contrary to expectations, fitness was not reduced in wild-type females mated to dj-1β males. Fecundity and fertility duration were increased in AOX and decreased in dj-1β females, indicating that female ROS levels may affect aging rates in stored sperm and/or eggs. Finally, we found evidence that accelerated aging in dj-1β sperm may have selected for more frequent mating. Our results help to clarify the relative roles of ROS production and ROS scavenging in the male and female reproductive systems.

scholarly journals Reactive oxygen species rescue regeneration after silencing the MAPK–ERK signaling pathway in Schmidtea mediterranea

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
V. Jaenen ◽  
S. Fraguas ◽  
K. Bijnens ◽  
M. Heleven ◽  
T. Artois ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Signaling Pathway ◽  
Reactive Oxygen ◽  
Light Therapy ◽  
Erk Signaling ◽  
Model Organisms ◽  
Ros Production ◽  
Oxygen Species ◽  
Planarian Regeneration ◽  
Egfr Signaling Pathway

AbstractDespite extensive research on molecular pathways controlling the process of regeneration in model organisms, little is known about the actual initiation signals necessary to induce regeneration. Recently, the activation of ERK signaling has been shown to be required to initiate regeneration in planarians. However, how ERK signaling is activated remains unknown. Reactive Oxygen Species (ROS) are well-known early signals necessary for regeneration in several models, including planarians. Still, the probable interplay between ROS and MAPK/ERK has not yet been described. Here, by interfering with major mediators (ROS, EGFR and MAPK/ERK), we were able to identify wound-induced ROS, and specifically H2O2, as upstream cues in the activation of regeneration. Our data demonstrate new relationships between regeneration-related ROS production and MAPK/ERK activation at the earliest regeneration stages, as well as the involvement of the EGFR-signaling pathway. Our results suggest that (1) ROS and/or H2O2 have the potential to rescue regeneration after MEK-inhibition, either by H2O2-treatment or light therapy, (2) ROS and/or H2O2 are required for the activation of MAPK/ERK signaling pathway, (3) the EGFR pathway can mediate ROS production and the activation of MAPK/ERK during planarian regeneration.

Sign in / Sign up

Export Citation Format

Share Document