scholarly journals Cold-Shock Test Is a Practical Method for Selecting Boar Ejaculates Yielding Appropriate Seminal Plasma for Post-Thawing Supplementation

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 871
Author(s):  
Estíbaliz Lacalle ◽  
Andrea Núñez ◽  
Estela Fernández-Alegre ◽  
Itxaso Crespo-Félez ◽  
Juan Carlos Domínguez ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Cold Shock ◽  
Practical Method ◽  
Sperm Viability ◽  
Disulfide Bridges ◽  
Kinematic Parameters ◽  
Shock Test ◽  
Progressive Motility ◽  
Neat Semen

Artificial insemination (AI) with cryopreserved semen is still unreliable for extensive pig industry application. Adding seminal plasma (SP) could improve post-thawing quality, but its suitability could vary. We applied a simple cold-shock test (CST, 5 min at 0 °C) on neat semen for classifying ejaculates (n = 63) as resistant or sensitive, obtaining two SP pools (CST-resistant: SPr, sensitive: SPs). Subsequently, frozen/thawed spermatozoa from six boars were incubated (37 °C) in MR-A® extender (control), 20% SPr, or 20% SPs, and analyzed at 0, 2, and 4 h. SP improved total and progressive motility, with a higher effect for SPr and STR (p < 0.05), decreasing kinematic parameters VCL and VAP, ALH, and BCF. Sperm viability was unaffected. SP increased apoptotic and membrane disorder ratios, and acrosomal damage, not affecting the chromatin structure (DNA fragmentation and immaturity by SCSA), protamination (CMA3), or disulfide levels (mBBr). However, the proportion of spermatozoa with elevated free thiols (disulfide bridges reduction) significantly increased. Results support a stimulatory role of SP on thawed semen, with additional benefits from SPr. The effect of SP and especially SPr after AI should be tested since CST could be a practical test for selecting suitable ejaculates in AI centers.

scholarly journals Effect of egg yolk-based extender and seminal plasma removal on sperm viability of cooled donkey semen

2021 ◽  
Vol 58 ◽  
pp. e174301
Author(s):  
Carolina Natalia Alonso ◽  
Catalina Castañeira ◽  
Ana Flores Bragulat ◽  
Luis Losinno
Keyword(s):  
Kinetic Parameters ◽  
Seminal Plasma ◽  
Membrane Integrity ◽  
Milk Proteins ◽  
Egg Yolk ◽  
Reproductive Efficiency ◽  
Sperm Viability ◽  
Progressive Motility ◽  
Live Sperm

Developing effective cooled semen protocols is essential to increase pregnancy rates and reproductive efficiency in donkeys. This study aimed to evaluate the effect on sperm kinetic parameters and membrane integrity in cooled donkey semen diluted with defined milk proteins extender with 1% or 2% of egg yolk and the removal of seminal plasma. Twenty-four ejaculates from six jackasses were collected. Each ejaculate was divided into four aliquots that were diluted in extender with 1% (EY1) or 2% (EY2) egg yolk. One sample from each group was centrifuged, seminal plasma was removed (CEY1, CEY2 groups, respectively), and the samples were then refrigerated at 5 °C for 24 h. Fresh and cooled semen samples were assessed for sperm motility, morphology, and plasma membrane integrity. Total motility, progressive motility, sperm kinetic parameters, or live sperm cells were not statistically different when semen was cooled with an extender supplemented with 1% or 2% of egg yolk. Seminal plasma removal does not affect total motility or sperm kinetic parameters. However, progressive motility decreased (P<0.05) when semen was extended with 2% of egg yolk and seminal plasma was removed. Membrane integrity was affected (P<0.05) in centrifuged samples. In conclusion, the obtained results suggest that there is no difference in sperm kinetics and membrane integrity when 1% or 2% of egg yolk was added to the Equiplus(R) extender. Also, the removal of seminal plasma by centrifugation did not have any beneficial effect on cooled donkey semen. Further studies are needed to relate these results with in vivo fertility tests with cooled donkey semen.

scholarly journals Cyclic AMP efflux through MRP4 regulates actin dynamics signalling pathway and sperm motility in bovines

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Nicolás Chiarante ◽  
Carlos A. I. Alonso ◽  
Jessica Plaza ◽  
Raquel Lottero-Leconte ◽  
Camila Arroyo-Salvo ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Actin Polymerization ◽  
Actin Dynamics ◽  
Kinematic Parameters ◽  
Progressive Motility ◽  
Bovine Sperm ◽  
Bovine Spermatozoa ◽  
Extracellular Camp ◽  
Kinematics Parameters

Abstract Previously we demonstrated that multidrug resistance-associated protein 4 transporter (MRP4) mediates cAMP efflux in bovine spermatozoa and that extracellular cAMP (ecAMP) triggers events associated to capacitation. Here, we deepen the study of the role of MRP4 in bovine sperm function by using MK571, an MRP4 inhibitor. The incubation of spermatozoa with MK571 during 45 min inhibited capacitation-associated events. MRP4 was localized in post-acrosomal region and mid-piece at 15 min capacitation, while at 45 min it was mainly located in the acrosome. After 15 min, MK571 decreased total sperm motility (TM), progressive motility (PM) and several kinematic parameters. The addition of ecAMP rescued MK571 effect and ecAMP alone increased the percentage of motile sperm and kinematics parameters. Since actin cytoskeleton plays essential roles in the regulation of sperm motility, we investigated if MRP4 activity might affect actin polymerization. After 15 min capacitation, an increase in F-actin was observed, which was inhibited by MK571. This effect was reverted by the addition of ecAMP. Furthermore, ecAMP alone increased F-actin levels while no F-actin was detected with ecAMP in the presence of PKA inhibitors. Our results support the importance of cAMP efflux through MRP4 in sperm capacitation and suggest its involvement in the regulation of actin polymerization and motility.

scholarly journals Retrograde flushing collection and freezing of dromedary camel epididymal spermatozoa with seminal plasma

2020 ◽  
Author(s):  
Davide Monaco ◽  
Miguel Batista ◽  
Olga Amann ◽  
Barbara Padalino ◽  
Wouter Pieters ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Egg Yolk ◽  
Dromedary Camel ◽  
Kinematic Parameters ◽  
Progressive Motility ◽  
Excellent Quality ◽  
Freezing Medium ◽  
Epididymal Spermatozoa

AbstractThe objectives of this study were to describe the parameters of dromedary camel epididymal spermatozoa collected by retrograde flushing (RF) technique and to evaluate the freezability of the collected sperm, diluted with and without the supplementation of seminal plasma (SP). Two experiments were conducted: in Experiment 1, ES were recovered within 6–8 h after castration; selected samples were diluted with a Tris-citrate egg-yolk glycerolated buffer and frozen. In Experiment 2, epididymides were stored for 24 h at 4 °C before RF and semen samples were frozen after dilution with a Tris-lactose egg-yolk glycerolated extender with and without 15% SP. In Experiment 1, eight semen samples were obtained from ten epididymides with a mean of 500 × 106 total spermatozoa recovered, per flushed epididymis. Mean post-thaw motility and progressive motility were 75 and 17%, respectively. In Experiment 2, 15 samples were collected, out of the 18 epididymides (mean number of collected spermatozoa: 700 × 106), and 13 of these samples were of excellent quality. Post-thaw parameters were not satisfactory but the supplementation of the freezing medium with 15% SP improved the progressive motility and kinematic parameters of the spermatozoa.

The role of differential phase contrast in electron microscopy

1978 ◽  
Vol 36 (1) ◽  
pp. 176-177
Author(s):  
E.M. Waddell ◽  
J.N. Chapman ◽  
R.P. Ferrier
Keyword(s):  
Phase Contrast ◽  
Practical Method ◽  
Position Vector ◽  
Differential Phase ◽  
Pure Phase ◽  
Differential Phase Contrast ◽  
The Difference ◽  
Image Position ◽  
First Moment

Dekkers and de Lang (1977) have discussed a practical method of realising differential phase contrast in a STEM. The method involves taking the difference signal from two semi-circular detectors placed symmetrically about the optic axis and subtending the same angle (2α) at the specimen as that of the cone of illumination. Such a system, or an obvious generalisation of it, namely a quadrant detector, has the characteristic of responding to the gradient of the phase of the specimen transmittance. In this paper we shall compare the performance of this type of system with that of a first moment detector (Waddell et al.1977).For a first moment detector the response function R(k) is of the form R(k) = ck where c is a constant, k is a position vector in the detector plane and the vector nature of R(k)indicates that two signals are produced. This type of system would produce an image signal given bywhere the specimen transmittance is given by a (r) exp (iϕ (r), r is a position vector in object space, ro the position of the probe, ⊛ represents a convolution integral and it has been assumed that we have a coherent probe, with a complex disturbance of the form b(r-ro) exp (iζ (r-ro)). Thus the image signal for a pure phase object imaged in a STEM using a first moment detector is b2 ⊛ ▽ø. Note that this puts no restrictions on the magnitude of the variation of the phase function, but does assume an infinite detector.

scholarly journals THE ROLE OF ZINC IN PROMOTING THE OPALESCENCE AND COLD-PRECIPITATION OF BOAR SEMINAL PLASMA

Reproduction ◽  
1974 ◽  
Vol 36 (1) ◽  
pp. 91-99 ◽  
Author(s):  
J. C. BOURSNELL ◽  
T. K. ROBERTS
Keyword(s):  
Seminal Plasma

The effects of Pb on sperm parameters and sperm DNA fragmentation of men investigated for infertility

2020 ◽  
Vol 31 (4) ◽  
Author(s):  
G.U.S. Wijesekara ◽  
D.M.S. Fernando ◽  
S. Wijeratne
Keyword(s):  
Dna Fragmentation ◽  
Seminal Plasma ◽  
Sperm Concentration ◽  
Sperm Parameters ◽  
World Health ◽  
Sperm Dna Fragmentation ◽  
Progressive Motility ◽  
Sperm Dna ◽  
The Mean ◽  
Pb Concentration

AbstractBackgroundLead (Pb) is one of the metals most prevalent in the environment and is known to cause infertility and deoxyribonucleic acid (DNA) fragmentation. This study aimed to determine the association between seminal plasma Pb and sperm DNA fragmentation in men investigated for infertility.MethodsMale partners (n = 300) of couples investigated for infertility were recruited after informed consent was obtained. Sperm parameters were assessed according to the World Health Organization (WHO) guidelines. Seminal plasma Pb was estimated by atomic absorption spectrophotometry after digestion with nitric acid.ResultsIn Pb-positive and -negative groups the sperm parameters and sperm DNA fragmentation were compared using independent sample t-test and the Mann-Whitney U-test, respectively. The mean [standard deviation (SD)] age and duration of infertility were 34.8 (5.34) years and 45.7 (35.09) months, respectively, and the mean Pb concentration was 15.7 μg/dL. In Pb positives compared to Pb negatives the means (SD) of sperm count, progressive motility viability and normal morphology were lower (p > 0.05) but the DNA fragmentation was significantly higher 39.80% (25.08) than Pb negatives 22.65% (11.30). Seminal plasma Pb concentration and sperm DNA fragmentation had a positive correlation (r = 0.38, p = 0.03). A negative correlation was observed between sperm DNA fragmentation and sperm concentration, progressive motility, total motility and viability. When the DNA fragmentation was ≥30% sperm concentration and viability decreased (p < 0.05).ConclusionsPb in seminal plasma had a significant effect on sperm DNA fragmentation but not with other sperm parameters.

scholarly journals Sperm Cryopreservation in American Flamingo (Phoenicopterus Ruber): Influence of Cryoprotectants and Seminal Plasma Removal

Animals ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 203
Author(s):  
María Gemma Millán de la Blanca ◽  
Eva Martínez-Nevado ◽  
Cristina Castaño ◽  
Juncal García ◽  
Berenice Bernal ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Genetic Material ◽  
First Year ◽  
Sperm Cryopreservation ◽  
Straight Line ◽  
The Family ◽  
Second Year ◽  
American Flamingo ◽  
Phoenicopterus Ruber

The American flamingo is a useful model for the development of successful semen cryopreservation procedures to be applied to threatened related species from the family Phoenicopteridae, and to permit genetic material banking. Current study sought to develop effective sperm cryopreservation protocols through examining the influences of two permeating cryoprotectants and the seminal plasma removal. During two consecutive years (April), semen samples were collected and frozen from American flamingos. In the first year, the effect of two permeating cryoprotectants, DMA (dimethylacetamide) (6%) or Me2SO (dimethylsulphoxide) (8%), on frozen–thawed sperm variables were compared in 21 males. No differences were seen between DMA and Me2SO for sperm motility, sperm viability, and DNA fragmentation after thawing. In the second year, the role of seminal plasma on sperm cryoresistance was investigated in 31 flamingos. Sperm samples were cryopreserved with and without seminal plasma, using Me2SO (8%) as a cryoprotectant. The results showed that samples with seminal plasma had higher values than samples without seminal plasma for the following sperm variables: Straight line velocity (22.40 µm/s vs. 16.64 µm/s), wobble (75.83% vs. 69.40%), (p < 0.05), linearity (62.73% vs. 52.01%) and straightness (82.38% vs. 73.79%) (p < 0.01); but acrosome integrity was lower (55.56% vs. 66.88%) (p < 0.05). The cryoresistance ratio (CR) was greater in samples frozen with seminal plasma than without seminal plasma for CR-progressive motility (138.72 vs. 54.59), CR-curvilinear velocity (105.98 vs. 89.32), CR-straight line velocity (152.77 vs. 112.58), CR-average path velocity (122.48 vs. 98.12), CR-wobble (111.75 vs. 102.04) (p < 0.05), CR-linearity (139.41 vs. 113.18), and CR-straightness (124.02 vs. 109.97) (p < 0.01). This research demonstrated that there were not differences between Me2SO and DMA to successful freezing sperm of flamingos; seminal plasma removal did not provide a benefit for sperm cryopreservation.

scholarly journals Viability of Najawa carp (Cyprinus carpio L.) sperm at 4°C temperature storage

2020 ◽  
Vol 147 ◽  
pp. 01015
Author(s):  
Dimas Fendy Pradana ◽  
Ignatius Hardaningsih ◽  
Dini Wahyu Kartika Sari
Keyword(s):  
Low Temperature ◽  
Cyprinus Carpio ◽  
Sperm Viability ◽  
Low Temperature Storage ◽  
Progressive Motility ◽  
C Storage ◽  
Randomized Design ◽  
Significant Difference ◽  
Carp Cyprinus Carpio ◽  
Temperature Storage

The objectives of this study were to evaluate the sperm viability of Najawa carp (Cyprinus carpio L.) in cryopreservation pre-conditions at 4°C. The design used in this study was Complete Randomized Design with 4 treatments, BSS as a control, 10% DMSO, 0,2 M Sucrose, and 5% DMSO + 0,1 M Sucrose; each consist of three replications. The parameters observed were progressive motility of fresh sperm, diluted sperm before low temperature storage, and 2 hours; 3 hours; 4 hours; 5 hours; one day; one week; a month after 4°C storage. The data were analyzed by ANOVA. The data showed that there was no significant difference between treatment (P>0.05). The best viability was 40.56% of sperm motility which survive for one week, it was achieved by 5% DMSO + 0,1 M Sucrose.

scholarly journals Autosomal dominant familial neurohypophyseal diabetes insipidus caused by a novel missense mutation in AVP gene in a large Italian kindred

Endocrine ◽  
2021 ◽  
Author(s):  
Carlotta Marzocchi ◽  
Silvia Cantara ◽  
Alfonso Sagnella ◽  
Maria Grazia Castagna ◽  
Marco Capezzone
Keyword(s):  
Diabetes Insipidus ◽  
Missense Mutation ◽  
Autosomal Dominant ◽  
Three Dimensional ◽  
Rare Condition ◽  
Central Diabetes Insipidus ◽  
Disulfide Bridges ◽  
Italian Family ◽  
Avp Gene

Abstract Purpose Familial neurohypophysial diabetes insipidus (FNDI), commonly caused by autosomal dominant arginine vasopressin (AVP) mutations, is a rare condition in which vasopressin fails in regulating body’s level of water with final polyuria and polydipsia. Genetic testing in familial cases of FNDI should be carry out to ensure adequate treatments and avoid disease manifestations especially in infants. Methods In this study, we investigated three-generations of a large Italian family with clinical diagnosis of familial central diabetes insipidus for the presence of potential pathogenic mutations in the AVP gene. Results We identified a heterozygous missense mutation (c.154 T > A; p.C52S) in AVP gene in all affected members studied of a large Italian family. In silico tools were used to investigate the pathogenic role of the mutation and three-dimensional protein structure predicted that the p.C52S impairs disulfide bridges formation resulting in misfolding of the protein. Conclusions This is the first study that identified a novel missense p.C52S mutation as causative of central diabetes insipidus in a large Italian pedigree.

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