scholarly journals Effects of cryopreservation on sperm quality, nuclear DNA integrity, in vitro fertilization, and in vitro embryo development in the mouse

Reproduction ◽  
2007 ◽  
Vol 133 (3) ◽  
pp. 585-595 ◽  
Author(s):  
Cengiz Yildiz ◽  
Palma Ottaviani ◽  
Napoleon Law ◽  
Renise Ayearst ◽  
Ling Liu ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Embryo Development ◽  
Membrane Integrity ◽  
Fertilization Rate ◽  
Development Rate ◽  
Dna Integrity ◽  
Plasma Membrane Integrity ◽  
Mouse Sperm ◽  
Progressive Motility

Efficient freezing, archiving, and thawing of sperm are essential techniques to support large scale research programs using mouse models of human disease. The purpose of this study was to investigate the effects of variable combinations and concentrations of cryoprotectants on sperm-assessment parameters of frozen–thawed mouse sperm in order to optimize cryopreservation protocols. Sperm was frozen using combinations of 3% skim milk + 0.2 or 0.3 M nonpermeating raffinose with either permeating glucose, fructose, propylene glycol, ethylene glycol, glycerol, or sodium pyruvate in CD-1, C3FeB6F1/J, B6129SF1, C57BL/6NCrIBR, 129S/SvPaslco, and DBA/2NCrIBR mice. Sperm-assessment parameters included progressive motility, plasma membrane integrity (SYBR-14 + PI),in vitrofertilization rate, andin vitroembryo development rate to blastocyst. DNA content analysis of sperm was measured by the sperm chromatin structure assay (SCSA). 0.3 M raffinose with 0.1 M fructose significantly improved post-thaw sperm-assessment parameters for CD-1, C3B6F1, B6129SF1 mice (P< 0.05–0.01), whereas 0.2 M raffinose with 0.1 M glycerol or 0.1 M fructose enhanced sperm assessment values for C57BL/6 and 129S mice (P< 0.01), compared to 0.3 M raffinose alone. DNA fragmentation during cryopreservation was significantly increased in all strains evaluated when compared with fresh control sperm in a strain-dependent manner (P< 0.01). Supplementation with permeating glycerol or fructose to the cryoprotectant (CPA) solution showed a significant protective effect to DNA integrity when cryopreserving sperm from C57BL/6 and 129S mice. Damage to sperm DNA significantly decreased the rate ofin vitroembryo development to blastocyst in C57BL/6 mice. The type of monosaccharide sugar or polyols, CPA molarity, and combination of permeating and nonpermeating cryoprotectant are significant factors for improving progressive motility, plasma membrane integrity, DNA integrity,in vitrofertilization rate, andin vitroembryo development rate to blastocyst in cryopreserved mouse sperm.

scholarly journals In vitro effect of codeine on human sperm motility and DNA integrity

2021 ◽  
Author(s):  
Roland Eghoghosoa Akhigbe ◽  
Moses Agbomhere Hamed ◽  
Lydia Oluwatoyin Ajayi ◽  
Davinson Chuka Anyogu ◽  
Ayodeji Folorunsho Ajayi
Keyword(s):  
Oxidative Stress ◽  
Plasma Membrane ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Human Spermatozoa ◽  
World Health ◽  
Dna Integrity ◽  
Plasma Membrane Integrity ◽  
Stress Dependent

Abstract Purpose This study assessed the in vitro effect of codeine, a popular drug of abuse, on human spermatozoa motility, plasma membrane integrity, DNA integrity, and oxidative stress. Materials and Methods Semen samples were collected from fifteen healthy donors and conventional semen analysis was carried out per the guideline of the World Health Organization. Direct Swim-up technique was performed to obtain highly motile sperm. Samples were incubated at 34.5°C with different concentrations (0, 0.1, 1, 5 and 10 mM) of codeine. The non-exposed (0 mM) was used as the control group. Sperm motility and DNA integrity were assessed at 30, 60, and 90 minutes, while sperm membrane integrity and sperm 8-OHdG level were determined at 90 minutes. Results Codeine at any tested concentration significantly reduced sperm motility and plasma membrane integrity but increased sperm 8-OHdG level compared to the control in a time-dependent manner. Furthermore, codeine at 1, 5, and 10 mM markedly increased sperm DNA damage. In addition, correlation study showed that sperm 8OHdG level was negatively associated with sperm motility, plasma membrane integrity, and DNA integrity. Conclusions Codeine may impair human spermatozoa fertilization capacity by inducing sperm dysmotility and damage to the sperm plasma membrane and DNA through an oxidative stress-dependent mechanism.

scholarly journals Polystyrene nanoplastics and microplastics can act as Trojan horse carriers of benzo(a)pyrene to mussel hemocytes in vitro

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alberto Katsumiti ◽  
María Paula Losada-Carrillo ◽  
Marta Barros ◽  
Miren P. Cajaraville
Keyword(s):  
Plasma Membrane ◽  
Cell Viability ◽  
Phagocytic Activity ◽  
Membrane Integrity ◽  
Aquatic Organisms ◽  
Particle Aggregation ◽  
Ros Production ◽  
Plasma Membrane Integrity ◽  
Intracellular Fate

AbstractIn this work we studied the ability of polystyrene (PS) nanoplastics (NPs) and microplastics (MPs) to transfer benzo(a)pyrene (BaP) to mussel hemocytes and to produce toxic effects in vitro. For this, intracellular fate and toxicity of PS NPs (0.05 μm) and MPs (0.5 and 4.5 μm) alone or with BaP and of BaP alone were assessed. Particles of 0.05 and 0.5 µm largely aggregated in the exposure medium whereas presence of BaP reduced particle aggregation. Cells internalized PS NPs and MPs alone or with BaP and these were found inside and outside lysosomes, depending on their size. PS particles alone or with BaP were cytotoxic to hemocytes only at the highest concentrations tested. The same was true for most sublethal endpoints except for increased phagocytic activity provoked by NPs and 0.5 μm MPs at lower concentrations. Plastic particles appeared to be the main drivers for reduced plasma membrane integrity and increased phagocytic and lysosomal activities whereas BaP appeared to contribute more to reduced cell viability and phagocytosis and increased ROS production and genotoxicity. Overall, PS NPs and MPs can act as carriers of BaP to mussel hemocytes, rising concerns about risks plastics associated to pollutants may pose to aquatic organisms.

scholarly journals Efficiency of Commercial Egg Yolk-Free and Egg Yolk-Supplemented Tris-Based Extenders for Dromedary Camel Semen Cryopreservation

Animals ◽  
2019 ◽  
Vol 9 (11) ◽  
pp. 999 ◽  
Author(s):  
Ayman Abdel-Aziz Swelum ◽  
Islam M. Saadeldin ◽  
Hani Ba-Awadh ◽  
Mohsen G. Al-Mutary ◽  
Abdullah F. Moumen ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Dna Integrity ◽  
Computer Assisted ◽  
Dromedary Camel ◽  
Plasma Membrane Integrity ◽  
Semen Cryopreservation

This study compared the efficiency of commercial egg yolk-free (AndroMed, OPTIXcell) and egg yolk-supplemented (Triladyl, Steridyl) Tris-based extenders for semen cryopreservation in seven adult dromedary camels. The camel-specific extender SHOTOR was used as control. The collected semen samples were evaluated and diluted with SHOTOR, Triladyl, Steridyl, AndroMed, or OPTIXcell. The diluted semen was gradually cooled and equilibrated for two hours before liquid nitrogen freezing. Semen was evaluated prior to freezing and after freeze-thawing cycles for motility, kinetics, vitality, abnormality, plasma membrane integrity, and DNA fragmentation using computer-assisted sperm analysis. In pre-freezing evaluation, progressive sperm motility was higher in SHOTOR-diluted samples (21.54 ± 1.83) than in samples diluted with Steridyl, OPTIXcell, or AndroMed (15.76 ± 1.80, 17.43 ± 1.10, and 13.27 ± 1.07, respectively). Moreover, Triladyl and SHOTOR resulted in significantly (p < 0.05) better sperm vitality and DNA integrity than all other diluents, but Triladyl resulted in a significantly (p < 0.05) better plasma membrane integrity (87.77 ± 0.31) than SHOTOR (85.48 ± 0.58). In the post-thawing evaluation, Triladyl led to significantly (p < 0.05) higher sperm motility (38.63 ± 0.81%; p < 0.05) when compared to SHOTOR, Steridyl or AndroMed (35.09 ± 1.341%, 34.4 ± 0.84%, and 31.99 ± 1.48%, respectively), with OPTIXcell being the least efficient (28.39 ± 0.86%). Progressive sperm motility was the highest when using Triladyl. Post-thawing curvilinear, straight line and average path sperm velocities were highest with Triladyl and lowest with AndroMed. Triladyl led to the highest linearity coefficient and straightness sperm coefficient, while SHOTOR to the highest DNA and plasma membrane integrity. OPTIXcell and AndroMed resulted in poor post-thawing sperm vitality, while Steridyl was less efficient than Triladyl. The highest rate of sperm abnormalities was recorded with OPTIXcell and the lowest with SHOTOR or Triladyl. In conclusion, SHOTOR, Triladyl, Steridyl, AndroMed, and OPTIXcell can all be used for camel semen cryopreservation; however, SHOTOR and Triladyl provided the best post-thawing sperm quality. Based on our findings, Triladyl is the best commercially available extender for dromedary camel semen cryopreservation to date.

Optimization of sperm freezability in Bactrian camel using various dilution rates and equilibration times

Zygote ◽  
2019 ◽  
Vol 27 (6) ◽  
pp. 362-366
Author(s):  
Vahid Vahedi ◽  
Mohsen Mostafaei ◽  
Hossein Vaseghi Dodaran ◽  
Nemat Hedayat Evrigh
Keyword(s):  
Plasma Membrane ◽  
Factorial Design ◽  
Healthy Adult ◽  
Membrane Integrity ◽  
Bactrian Camel ◽  
Adult Males ◽  
Kinematic Parameters ◽  
Freezing And Thawing ◽  
Plasma Membrane Integrity ◽  
Progressive Motility

SummaryThe effect of different dilution rates and equilibration times on the cryopreservation of Bactrian camel spermatozoa was evaluated in the current study. Semen samples from four healthy adult males were collected, processed and pooled. They were then subjected to a completely randomized 4×2 factorial design including four dilution rates (DR; 1:1, 1:2, 1:4 or 1:8; v:v with SHOTOR diluent) and two equilibration times (ET; 1 or 2 h at 5ºC). After freezing and thawing, sperm kinematic parameters as well as viability, plasma membrane integrity, abnormality and seminal malondialdehyde level were assessed. According to the results, four-fold diluted samples recorded significantly higher values (P < 0.05) for sperm total (39.58 vs 31.83 and 33.33,%) and progressive motility (19.50 vs 14.00 and 14.25,%), viability (55.37 vs 43.50 and 48.75,%) and plasma membrane integrity (46.75 vs 37.25 and 37.37,%) than those of both less (1:1) and high (1:8) concentrated samples, respectively. By contrast, the percentage of abnormal spermatozoa and the concentration of seminal malondialdehyde were comparable among all treated groups. Moreover, ET revealed that 1 h equilibration had significantly higher sperm motility (37.04 vs 33.33%), linearity (42.29 vs 32.26%), beat cross-frequency (13.15 vs 8.70 Hz), plasma membrane integrity (42.25 vs 39.75%) and viability (51.37 vs 48.12%) compared with 2 h of ET (P < 0.05). Taken together, a four-fold dilution along with 1 h equilibration can be an optimal procedure to cryopreserve Bactrian camel sperm.

scholarly journals Antisperm antibodies disrupt plasma membrane integrity and inhibit tyrosine phosphorylation in human spermatozoa

2018 ◽  
Vol 27 (1) ◽  
pp. 3-11 ◽  
Author(s):  
Dwi A. Pujianto ◽  
Hajizah Hajizah ◽  
Indra G. Mansur ◽  
Amarudin Amarudin
Keyword(s):  
Plasma Membrane ◽  
Tyrosine Phosphorylation ◽  
Membrane Integrity ◽  
In Vitro Study ◽  
Dependent Manner ◽  
Sperm Viability ◽  
Plasma Membrane Integrity ◽  
Infertile Women ◽  
The Status

Background: The etiology of unexplained infertility has not been fully understood. This study aimed to determine the effect of antisperm antibody (ASA) from infertile women on viability, motility, plasma membrane integrity, and status of tyrosine phosphorylation in the human spermatozoa.Methods: An experimental in vitro study was conducted at the Department of Biology, Faculty of Medicine, Universitas Indonesia from February to November 2014. Spermatozoa from normal fertile donors was incubated with serum containing ASA from infertile women at several dilutions (1/1000, 1/100, 1/10, and without dilution) for 1 and 2 hours. The plasma membrane integrity was assessed with hypoosmotic swelling (HOS) test, whereas the status of tyrosine phosphorylation was analyzed using Western immunoblotting and immunocytochemistry.Results: After 1 hour incubation time, ASA caused a decrease in sperm viability, motility, plasma membrane integrity, and inhibit sperm tyrosine phosphorylation. ASA caused a decrease in viability, motility, sperm plasma membrane integrity, and tyrosine phosphorylation of sperm after 1 hour incubation time.Conclusion: ASA from infertile women reduced the sperm viability, motility, plasma membrane integrity, and capacitation in dose and time dependent manner.

scholarly journals Mouse sperm delivery via cauda epididymis without transport medium

2016 ◽  
Author(s):  
Man-Xi Jiang ◽  
Mei-Shan Wang ◽  
Xiang-Hong Ou ◽  
Xue-Jin Chen ◽  
Yan Zhu
Keyword(s):  
Embryo Development ◽  
Sperm Motility ◽  
Room Temperature ◽  
Sperm Quality ◽  
Mouse Sperm ◽  
Spermatozoa Motility ◽  
Transport Medium ◽  
Fertilization Rates ◽  
Progressive Motility

This study was aimed to investigate the effects of room temperature (RT, 20-25 oC) and absence of medium during cauda epididymis transport on spermatozoa quality, fertility and embryo development. In the first experiment, fresh sperm from one side of cauda epididymis was used for in vitro fertilization, and another side was delivered at RT or 4-8 oC either with or without M2. In the second experiment, each side of cauda epididymis obtained from the same mouse was individually delivered at RT or 4 oC with or without M2. Finally, sperm motility, progressive motility scores and fertility of fresh spermatozoa or those from transported cauda epididymis, and IVF embryo development were evaluated. Progressive motility scores and fertilization rates were higher in fresh spermatozoa than transported sperm; sperm motility of transported cauda epididymis at 4-8 oC was comparable to fresh spermatozoa, but spermatozoa motility of transported cauda epididymis at RT was inferior to fresh spermatozoa. Spermatozoa motillty of transported cauda epididymis at 4-8 oC with transport medium was much higher than that without transport medium; absence of transport medium did not affect sperm motility of transported cauda epididymis at 4-8 oC but affected sperm motility of transported cauda epididymis at RT. Sperm quality from transported cauda epididymis can be efficiently kept at 4-8 oC, and cauda epididymis transport at 4-8 oC without M2 is more beneficial on keeping their fertility. Moreover cauda epididymis transport at RT without medium could sufficiently produce embryos for obtainning live offsprings.

Characteristics of high-quality Asian elephant (Elephas maximus) ejaculates and in vitro sperm quality after prolonged chilled storage and directional freezing

2013 ◽  
Vol 25 (5) ◽  
pp. 790 ◽  
Author(s):  
J. K. O'Brien ◽  
K. J. Steinman ◽  
G. A. Montano ◽  
C. C. Love ◽  
R. L. Saiers ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Elephas Maximus ◽  
Mitochondrial Activity ◽  
Dna Integrity ◽  
High Quality ◽  
Chilled Storage ◽  
Progressive Motility ◽  
Directional Freezing

The in vitro quality of spermatozoa from one elephant (Elephas maximus) was examined after chilled storage and directional freezing (DF). High-quality, non-contaminated ejaculates (77.6 ± 6.0% progressive motility, 3.9 ± 1.5 µg creatinine mL–1 raw semen, 2.7 ± 0.6% detached heads) were cryopreserved after 0 (0hStor), 12 (12hStor) and 24 h (24hStor) of chilled storage. At 0 h and 6 h post-thawing, total motility, plasma membrane integrity, acrosome integrity, mitochondrial activity and normal morphology were similar (P > 0.05) across treatments. In contrast, progressive motility, rapid velocity and several kinematic parameters were lower (P < 0.05) for 24Stor compared with 0hStor at 0 h post-thaw. By 6 h post-thaw, amplitude of lateral head displacement and velocity parameters (average pathway, straight-line and curvilinear velocity) were lower (P < 0.05) for 24hStor compared with 0hStor and 12hStor. DNA integrity was high and remained unchanged (P > 0.05) across all groups and processing stages (1.6 ± 0.6% of cells contained fragmented DNA). Results indicate that DF after up to 12 h of chilled storage results in a post-thaw sperm population of acceptable quality for artificial insemination. These findings have implications for the cryopreservation of sex-sorted spermatozoa, which typically undergo more than 12 h of chilled storage prior to sorting and preservation.

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