309 DIFFERENCES IN PRONUCLEUS FORMATION RATES BETWEEN BULLS IN RESPONSE TO GAMMA IRRADIATION OF FROZEN - THAWED SEMEN

2007 ◽  
Vol 19 (1) ◽  
pp. 270
Author(s):  
M. M. Petersen ◽  
G. B. Boe-Hansen ◽  
A. Birck ◽  
B. Avery ◽  
M. Jensen ◽  
...  
Keyword(s):  
Dna Damage ◽  
Gamma Irradiation ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Fertilization Rate ◽  
High Dose ◽  
Exact Test ◽  
Bovine Spermatozoa ◽  
Gamma Irradiated

Gamma irradiation induces DNA damage to mature bovine spermatozoa but does not affect motility, membrane integrity, or function (Fatehi et al. 2006 J. Androl. 27, 176–188), making it a useful model to evaluate the effect of fertilization with DNA-damaged spermatozoa. The objective of the present study was to analyze the fertilizing capacity of frozen–thawed gamma-irradiated sperm from 2 bulls: a bull with a high sperm quality and fertilization rate (bull A), and a bull with a low sperm quality and fertilization rate (bull B). To ensure that DNA damage was induced, frozen–thawed semen was exposed to a high dose of irradiation (90 Gy). Cumulus–oocyte complexes were obtained from abattoir ovaries and were in vitro-matured (IVM) using standard procedures (23 h in DMEM with 5% serum and eCG/hCG). For each treatment group, 3 to 5 straws of semen from the same ejaculate were used. After thawing, semen from the 2 bulls was either irradiated or held at room temperature before 1:1 dilution in sperm-TALP. During irradiation, semen was kept in the straws. The irradiated and nonirradiated sperm were used for IVF (23 h in IVF-TALP) within 1 h of thawing. IVM and IVF were carried out at 38.5�C in 5% CO2 in air. After IVF, presumptive zygotes were whole mount-fixed and 24 h later were stained with 1% aceto-orcein followed by determination of fertilization status. Fertilization was considered normal if 2 pronuclei (PN) were present. A total of 4 replicates were performed and 286 zygotes analyzed. No difference was detected between replicates, and the results were pooled. Fisher's exact test was used to determine effect of treatment. More zygotes had 2 PN after fertilization with semen from bull A (47/67 = 70%), compared with that from bull B (16/71 = 23%; P < 0.0001) using nonirradiated sperm. Irradiation of sperm significantly increased the fraction of zygotes with 2 PN from bull A (61/70 = 87%; P = 0.03), but decreased the 2 PN fractions in zygotes fertilized with sperm from bull B (2/75 = 3%; P = 0.0002). Ideally, to avoid straw variation, semen should have been pooled and divided into groups before irradiation. The variation between straws might explain the higher fertilization rates in bull A using irradiated sperm compared with nonirradiated sperm. In conclusion, there appear to be differences in fertilizing ability between bulls after irradiation of frozen–thawed sperm. This could be due to suboptimal DNA packaging, which made sperm from bull B more susceptible to radiation-induced damage. The potential irradiation-induced increase in DNA fragmentation in sperm from bull B compared with that from bull A might delay or prevent the formation of the 2 PN. Further studies are needed to investigate differences in fertilization and early embryonic development using sperm with intact or damaged DNA.

scholarly journals Inhibitors of zinc-dependent metalloproteases hinder sperm passage through the cumulus oophorus during porcine fertilization in vitro

Reproduction ◽  
2012 ◽  
Vol 144 (6) ◽  
pp. 687-697 ◽  
Author(s):  
J Beek ◽  
H Nauwynck ◽  
D Maes ◽  
A Van Soom
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Inhibitory Effect ◽  
Fertilization Rate ◽  
Quality Parameters ◽  
Fertilization In Vitro ◽  
Cumulus Oophorus ◽  
Sperm Penetration

In this study, we report for the first time on a possible contribution of metalloproteases in sperm passage through the cumulus matrix in pigs. The presence of 20 μM 1,10-phenanthroline (1,10-PHEN), inhibitor of zinc-dependent metalloproteases, strongly inhibited the degree of sperm penetration in cumulus-intact (CI), but not in cumulus-free (CF), porcine oocytes during IVF. The inhibitory effect of 1,10-PHEN was due to the chelation of metal ions as a non-chelating analog (1,7-PHEN) did not affect IVF rates. Furthermore, incubation with 1,10-PHEN did not affect sperm binding to the zona pellucida nor sperm motility, membrane integrity, or acrosomal status. These findings led to the assumption that 1,10-PHEN interacts with a sperm- or cumulus-derived metalloprotease. Metalloproteases are key players in physiological processes involving degradation or remodeling of extracellular matrix. In vivo, their proteolytic activity is regulated by tissue inhibitors of metalloproteases (TIMP1–TIMP4). We tested the effect of TIMP3 on fertilization parameters after porcine IVF. Similar to 1,10-PHEN, TIMP3 inhibited total fertilization rate of CI but not CF oocytes and did not influence sperm quality parameters. Although the inhibitory effect was stronger in CI oocytes, TIMP3 also reduced the degree of sperm penetration in CF oocytes, suggesting the involvement of a metalloprotease in a subsequent step during fertilization. In conclusion, our results indicate the involvement of TIMP3-sensitive, zinc-dependent metalloprotease activity in sperm passage through the cumulus oophorus in pigs. The results should provide the basis for further biochemical research toward the localization and identification of the metalloprotease involved.

Seminal plasma proteins protect flow-sorted ram spermatozoa from freeze - thaw damage

2009 ◽  
Vol 21 (4) ◽  
pp. 571 ◽  
Author(s):  
T. Leahy ◽  
J. I. Marti ◽  
G. Evans ◽  
W. M. C. Maxwell
Keyword(s):  
Seminal Plasma ◽  
High Speed ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Weight Fraction ◽  
Protein Supplementation ◽  
Freezing Process ◽  
Spermatozoa Motility ◽  
Functional Integrity

Seminal plasma improves the functional integrity of compromised ram spermatozoa but has been reported to be toxic to sorted spermatozoa. The present study attempted to clarify this paradoxical effect and improve the functional integrity of spermatozoa following sorting and cryopreservation. The in vitro function of sorted spermatozoa (motility characteristics and membrane integrity) was examined after supplementation with differing concentrations and protein fractions of seminal plasma at various stages of the sorting and freezing process. For all experiments, spermatozoa (two males, n = four ejaculates per male) were processed through a high-speed flow cytometer before cryopreservation, thawing and incubation for 6 h (37°C). Supplementation of crude seminal plasma (CP), its low molecular weight fraction (LP; <10 kDa) or protein-rich fraction (SPP; >10 kDa), immediately before freezing improved the functional integrity of sorted spermatozoa compared with no supplementation (control), whereas supplementation after thawing had no effect for CP and LP. The protective effect of seminal plasma was not altered by increasing the amount of protein supplementation. No toxic effect of CP, SPP or LP was evident even when supplemented at high protein concentrations. It is concluded that seminal plasma protein, if added to ram spermatozoa after sorting and before freezing, can improve post-thaw sperm quality and consequently the efficiency of sorting. This effect is most likely related to protection of the spermatozoa during freeze–thawing.

Measurement of Alkali Labile DNA Damage and Cross-Links Following 2450 MHz Microwave and Low Dose Gamma Irradiation In Vitro

2006 ◽  
pp. 135-142
Author(s):  
I. Lagroye ◽  
B. Wettring ◽  
E. G. Moros ◽  
W. L. Straube ◽  
W. F. Pickard ◽  
...  
Keyword(s):  
Dna Damage ◽  
Gamma Irradiation ◽  
Low Dose ◽  
Cross Links

scholarly journals Preparation of cyclodextrin nanoparticles and evaluation of its effect on the capacitation of bovine spermatozoa used in the in vitro fertilization

2018 ◽  
Vol 9 (4) ◽  
pp. 112
Author(s):  
Heba F Salem ◽  
Mai Raslan ◽  
Hanaa Suliman ◽  
Tamer Essam ◽  
Saber Abd-Allah
Keyword(s):  
Data Analysis ◽  
In Vitro Fertilization ◽  
Interval Data ◽  
Fertilization Rate ◽  
Vitro Fertilization ◽  
Bovine Spermatozoa ◽  
The Media ◽  
Biomedical Interventions ◽  
Positive Effect

<p>This study was conducted to produce nanosized cyclodextrin (NCD) and assess its effect on bovine spermatozoa during In vitro fertilization (IVF) to optimize the capacitation media for successful IVF. Therefore, Four cyclodextrin formulations were prepared and characterized. Data analysis revealed the best formula (F2) showed a smallest particle size (15 nm), zeta potential (-37 mv), and higher yield percentages (95%) was selected for spem capacitation. Motile spermatozoa were separated from frozen-thawed semen by a swim-up procedure and capacitated in IVF-TALP medium with different formulae of NCD or CD or without treatments (control) and incubated for 3hours(hr) at 38°C and evaluated every one (hr) interval. Data analysis revealed that the formulation of cyclodextrin nanoparticles (F2<strong>)</strong> after (2hr) incubation in the media gave best effect on sperm capacitation and acrosme reaction (AR) and effect of sperm treated with NCD on fertilization rate was evaluated. The results showed that the proportion of Oocytes fertilized was increased significantly in F2 (60%) than in the control (35%), and cyclodextrin group (50%) groups (<em>p</em>&lt;0.05). It could be inferred from this investigation that cyclodextrin nanoparticles can be used for biomedical interventions in bovine spermatozoa. NCD improve sperm motility, viability, and (AR), also fertilization rate of sperm treated with NCD increase. So NCD gave positive effect on sperm functions during IVF. </p>

Antinutritional factors, ruminal degradation and in vitro protein digestibility of gamma irradiated full-fat soybean

2009 ◽  
Vol 2009 ◽  
pp. 184-184
Author(s):  
M Taghinejad Roudbaneh ◽  
A Nikkhah ◽  
A A Sadeghi ◽  
G Raisali ◽  
M Chamani
Keyword(s):  
Gamma Irradiation ◽  
Dairy Cows ◽  
Nutritional Value ◽  
Protein Digestibility ◽  
High Energy ◽  
Antinutritional Factors ◽  
Γ Irradiation ◽  
Rumen Microbes ◽  
Gamma Irradiated

Whole soybean is used as a high energy-protein supplement for dairy cows, but contains various antinutritional factors and the protein is highly degradable by rumen microbes. Roasting and extrusion are very popular ways of feeding soybeans to dairy cows but these processing methods may adversely affect the protein digestibility and lysine availability of the final product in the small intestine. Gamma irradiation has been recognized as a reliable and safe method for improving the nutritional value and inactivation or removal of certain antinutritional factors in foods and feeds (Siddhuraju et al., 2002). As far as we know, information about the effects of γ-irradiation on ruminal crude protein (CP) degradation of full-fat soybean in the literature is scarce. The aim of this study was to evaluate the effects of gamma irradiation (γ-irradiation) on nutritional value and antinutritional factors of soybean (SB) for ruminants.

scholarly journals Validation of the Turkey Semen Cryopreservation by Evaluating the Effect of Two Diluents and the Inseminating Doses

Animals ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 1329
Author(s):  
Michele Di Iorio ◽  
Giusy Rusco ◽  
Roberta Iampietro ◽  
Lucia Maiuro ◽  
Achille Schiavone ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Sperm Concentration ◽  
Membrane Integrity ◽  
In Vivo Test ◽  
Sperm Analysis ◽  
Fertilizing Ability ◽  
Vitro Analysis ◽  
Hatching Rates

This study was designed to test the fertilizing ability of cryopreserved turkey semen, and here, two experiments were performed: an in vitro analysis to assess the effects of Tselutin and Lake diluents and an in vivo test to determine the fertility and hatching rates by also studying the feat of three insemination doses (250, 400 and 600 × 106 sperm/hen). Pooled semen samples were diluted with Tselutin or Lake extender which contained 20% of dimethylsulfoxide and 1 mM of Ficoll at final sperm concentration of 3 × 109 sperm/mL. Thereafter, semen was packaged into straws and frozen on liquid nitrogen. The post-thaw sperm quality was evaluated considering motility (computer-aided sperm analysis—CASA system) and membrane integrity (flow cytometry). Significantly higher values of progressive motility and some kinetic parameters in semen frozen with Lake were found. When we compared the extenders in vivo, no significant effects were detected, whilst sperm concentration significantly affected both fertility and hatching rates, with the best results obtained with the sperm concentration of 400 × 106 sperm/hen. From the results obtained, it emerged that the extender type only affected sperm motility characteristics, not the fertilizing ability of frozen-thawed semen, while inseminating dose markedly affected fertility and hatching rates.

scholarly journals O papel do dimetilsulfóxido na criopreservação de sêmen de Curimba (Prochilodus lineatus)

2015 ◽  
Vol 36 (5) ◽  
pp. 3471
Author(s):  
Antonio Sergio Varela Junior ◽  
Estela Fernandes Silva ◽  
Tainã Figueiredo Cardoso ◽  
Érica Yokoyama Namba ◽  
Rodrigo Desessards Jardim ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Fertilization Rate ◽  
Dna Integrity ◽  
Hatching Rate ◽  
Sperm Viability ◽  
Prochilodus Lineatus ◽  
Dmso Concentration ◽  
Fresh Semen

<p class="Pa7">Cryopreservation of Curimba semen (Prochilodus lineatus) is ecological and commercial importance. The objective of this study was to evaluate the effect of different concentrations (2, 5, 8 and 11%) of dimethyl sulfoxide (DMSO) diluted in Betsville Thawing Solution (BTS) on the quality of post-thaw semen Curimba. We analyzed the rate and period motility, sperm viability, membrane integrity and DNA, mitochondrial functionality, and fertilization and hatching rate. The plasma membrane and DNA integrity of a DMSO concentration of 11% obtained better results than the concentration of 5% (p &lt;0.05). However, treatment of 5% DMSO resulted in a longer latency and a higher fertilization rate and hatching, in other sperm quality equal to that of fresh semen. The results of this study indicate that 5% DMSO is ideal for cryopreservation of semen Curimba.</p>

scholarly journals Evaluation of sperm quality of Erythrolamprus poecilogyrus sublineatus (Cope, 1860) (Serpentes, Dipsadidae)

2017 ◽  
Vol 77 (3) ◽  
pp. 553-557
Author(s):  
A. C. Silva ◽  
A. S. Varela Junior ◽  
T. F. Cardoso ◽  
E. F. Silva ◽  
D. Loebmann ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Coastal Region ◽  
Rio Grande ◽  
Rio Grande Do Sul ◽  
Sperm Analysis ◽  
Pampa Region

Abstract Erythrolamprus poecilogyrus sublineatus (Cope, 1860), is a species widely distributed in the Pampa Domain, occurring in Rio Grande do Sul, Argentina and Uruguay, mainlyin the pampa region. In the coastal region of southern Brazil this is serpent is considered one of the most abundant. The purpose of the present study is to describe the techniques of sperm evaluation in vitro for E. poecilogyrus sublineatus in the coastal plain of Rio Grande do Sul, Brazil. After laparatomy the efferent vases were collected and the semen was diluted in 1ml Beltsville Thawing Solution. The characteristics of motility, membrane integrity, mitochondria, acrosome, DNA, cell viability and cellular functionality were evaluated. Fluorescent probes were used for the evaluation of sperm structure in epifluorescence microscope. With the techniques described, it was possible to identify intact and injured cells, enabling the determination of cell characteristics for the spring season (October and November). It was observed in the analyses that 80% of sperm cells were mobile and that 84.1 ± 8.0% of sperm membranes were intact. The standards found were of 48 ± 13.8% of intact acrosome, 73.6 ± 6.0 of perfect DNA and of 91.8 ± 4.0 of functional mitochondria. Thus, these values from the sperm analysis can be used as standards for the species Erythrolamprus poecilogyrus sublineatus.

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 Stimulatory Effects of Gamma Irradiation on Phytochemical Properties, Mitotic Behaviour, and Nutritional Composition of Sainfoin (Onobrychis viciifoliaScop.)

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Sadegh Mohajer ◽  
Rosna Mat Taha ◽  
Ma Ma Lay ◽  
Arash Khorasani Esmaeili ◽  
Mahsa Khalili
Keyword(s):  
Gamma Irradiation ◽  
Nutritional Composition ◽  
Anthocyanin Content ◽  
Irradiation Intensity ◽  
Dry Matter Digestibility ◽  
Chromosome Bridge ◽  
Hplc Profiles ◽  
Positive Correlations ◽  
Gamma Irradiated

Sainfoin (Onobrychis viciifoliaScop. Syn.Onobrychis sativaL.) is a bloat-safe forage crop with high levels of tannins, which is renowned for its medicinal qualities in grazing animals. Mutagenesis technique was applied to investigate the influence of gamma irradiation at 30, 60, 90, and 120 Gy on mitotic behavior,in vitrogrowth factors, phytochemical and nutritional constituents of sainfoin. Although a percentage of plant necrosis and non-growing seed were enhanced by irradiation increment, the germination speed was significantly decreased. It was observed that gamma irradiated seeds had higher value of crude protein and dry matter digestibility compared to control seeds. Toxicity of copper was reduced in sainfoin irradiated seeds at different doses of gamma rays. Anthocyanin content also decreased in inverse proportion to irradiation intensity. Accumulation of phenolic and flavonoid compounds was enhanced by gamma irradiation exposure in leaf cells. HPLC profiles differed in peak areas of the two important alkaloids, Berberine and Sanguinarine, in 120 Gy irradiated seeds compared to control seeds. There were positive correlations between irradiation dose and some abnormality divisions such as laggard chromosome, micronucleus, binucleated cells, chromosome bridge, and cytomixis. In reality, radiocytological evaluation was proven to be essential in deducing the effectiveness of gamma irradiation to induce somaclonal variation in sainfoin.

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