Detection of intracellular reactive oxygen species (superoxide anion and hydrogen peroxide) and lipid peroxidation during cryopreservation of alpaca spermatozoa

Evangelista‐Vargas, Shirley; Santiani, Alexei

doi:10.1111/rda.12984

Cited by 30 publications

(34 citation statements)

References 27 publications

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“…Our results of

O_{2}^{-}

in fresh spermatozoa are consistent with those reported in previous

O_{2}^{-}

studies of boar (Guthrie, Welch, & Long, ), equine (Morrell et al, ) and human spermatozoa (Mahfouz, Sharma, Lackner, Aziz, & Agarwal, ), in semen of good quality. The study revealed that percentages sperm positive with

O_{2}^{-}

significantly increased after cryopreservation of cat spermatozoa, as is previously described for human (Aitken & Krausz, ; De Iuliis et al, ), bovine (Ahmad et al, ; Bilodeau, Chatterjee, Sirard, & Gagnon, ; Chatterjee & Gagnon, ; O'Flaherty, Beorlegui, & Beconi, ), alpaca (Evangelista‐Vargas & Santiani, ) and equine spermatozoa (Yeste et al, ).…”

Section: Discussionsupporting

confidence: 82%

Changes in sperm function and structure after freezing in domestic cat spermatozoa

et al. 2018

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Sperm cryopreservation allows for a long-term storage of genetic. However, changes due to factors as cold shock, osmotic and oxidative stress cause reduction in viability and fertilising ability of frozen/thawed spermatozoa. Therefore, evaluation of cryoinjury of cat spermatozoa is a key factor in achieving better cryopreservation results. This study analysed the changes in structural and functional after freezing in ejaculated domestic cats spermatozoa. Semen samples (n = 60) were analysed before and after freezing, progressive motility was determined with computer-assisted sperm analysis and viability, and acrosome intact spermatozoa, mitochondrial function and superoxide anion ( ) were assessed by flow cytometry. The results demonstrated that cryopreservation induced changes in all sperm parameters (p< 0.05). Total sperm motility, viability, acrosome integrity and mitochondrial function of fresh samples were near to 80% and decrease near to 40% in frozen/thawed spermatozoa (p < 0.05); nevertheless, in contrast to all other sperm parameters, the sperm positive with increased post/thawing (p< 0.05). In conclusion, changes in frozen/thawed spermatozoa could be related to the effect of oxidative stress due to the increase in the synthesis of and a concomitant loss of functional competence. Therefore, the evaluation of these sperm parameters could contribute to complement the analysis of fresh or frozen semen used ART.

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“…Our results of

O_{2}^{-}

in fresh spermatozoa are consistent with those reported in previous

O_{2}^{-}

O_{2}^{-}

Section: Discussionsupporting

confidence: 82%

Changes in sperm function and structure after freezing in domestic cat spermatozoa

et al. 2018

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“…In support of these findings, LPO was not related to freezing-thawing of fresh and frozen human sperm (36). In rams, LPO did not change during cooling and thawing procedures of cryopreservation (37). Moreover, young and old buffalo bulls had similar levels of lipid peroxidation in fresh vs. frozen-thawed sperm (38).…”

Section: Discussionsupporting

confidence: 54%

Cellular and Functional Physiopathology of Bull Sperm With Altered Sperm Freezability

et al. 2020

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The objective of this study was to ascertain the cellular and functional parameters as well as ROS related changes in sperm from bulls with varied sperm freezability phenotypes. Using principal component analysis (PCA), the variables were reduced to two principal components, of which PC1 explained 48% of the variance, and PC2 explained 24% of the variance, and clustered animals into two distinct groups of good freezability (GF) and poor freezability (PF). In ROS associated pathophysiology, there were more dead superoxide anion positive (Dead SO+) sperm in GF bulls than those in PF (15.72 and 12.00%; P = 0.024), and that Dead SO+ and live hydrogen positive cells (live H 2 O 2 +) were positively correlated with freezability, respectively (R 2 = 0.55, P < 0.0130) and (r s = 0.63, P = 0.0498). Related to sperm functional integrity, sperm from PF bulls had greater dead intact acrosome (DIAC) than those from GF bulls (26.29 and 16.10%; P = 0.028) whereas sperm from GF bulls tended to have greater live intact acrosome (LIAC) than those from PF bulls (64.47 and 50.05%; P = 0.084). Sperm with dead reacted acrosome (DRAC) in PF bulls were greater compared to those in GF (19.27 and 11.48%; P = 0.007). While DIAC (R 2 = 0.56, P = 0.0124) and DRAC (R 2 = 0.57, P < 0.0111) were negatively correlated with freezability phenotype, LIAC (R 2 = 0.36, P = 0.0628) was positively correlated. Protamine deficiency (PRM) was similar between sperm from GF and PF bulls (7.20 and 0.64%; P = 0.206) and (r s = 0.70, P = 0.0251) was correlated with freezability. Sperm characteristics associated with cryotolerance are important for advancing both fundamental andrology and assisted reproductive technologies across mammals.

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“…The production of intracellular superoxide was measured by the fluorescence probe dihydroethidium (DHE, Invitrogen Life Technologies, Grand Island, NY, USA) as described previously [31] . Briefly, cells were incubated with paraquat and maneb overnight in 24-well plates, and then were incubated with 10 μM DHE for 30 mins at 37 °C.…”

Section: Methodsmentioning

confidence: 99%

Complement receptor 3 mediates NADPH oxidase activation and dopaminergic neurodegeneration through a Src-Erk-dependent pathway

et al. 2018

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Microglial NADPH oxidase (Nox2) plays a key role in chronic neuroinflammation and related dopaminergic neurodegeneration in Parkinson's disease (PD). However, the mechanisms behind Nox2 activation remain unclear. Here, we revealed the critical role of complement receptor 3 (CR3), a microglia-specific pattern recognition receptor, in Nox2 activation and subsequent dopaminergic neurodegeneration by using paraquat and maneb-induced PD model. Suppression or genetic deletion of CR3 impeded paraquat and maneb-induced activation of microglial Nox2, which was associated with attenuation of dopaminergic neurodegeneration. Mechanistic inquiry revealed that blocking CR3 reduced paraquat and maneb-induced membrane translocation of Nox2 cytosolic subunit p47phox, an essential step for Nox2 activation. Src and Erk (extracellular regulated protein kinases) were subsequently recognized as the downstream signals of CR3. Moreover, inhibition of Src or Erk impaired Nox2 activation in response to paraquat and maneb co-exposure. Finally, we found that CR3-deficient mice were more resistant to paraquat and maneb-induced Nox2 activation and nigral dopaminergic neurodegeneration as well as motor dysfunction than the wild type controls. Taken together, our results showed that CR3 regulated Nox2 activation and dopaminergic neurodegeneration through a Src-Erk-dependent pathway in a two pesticide-induced PD model, providing novel insights into the immune pathogenesis of PD.

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Detection of intracellular reactive oxygen species (superoxide anion and hydrogen peroxide) and lipid peroxidation during cryopreservation of alpaca spermatozoa

Cited by 30 publications

References 27 publications

Changes in sperm function and structure after freezing in domestic cat spermatozoa

Changes in sperm function and structure after freezing in domestic cat spermatozoa

Cellular and Functional Physiopathology of Bull Sperm With Altered Sperm Freezability

Complement receptor 3 mediates NADPH oxidase activation and dopaminergic neurodegeneration through a Src-Erk-dependent pathway

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