71 BOVINE SPERM DEATH KINETICS: CHANGES IN MOTILITY, ACROSOMES, AND PLASMA MEMBRANE

2013 ◽  
Vol 25 (1) ◽  
pp. 183
Author(s):  
M. Ahmad ◽  
N. Ahmad ◽  
A. Riaz ◽  
M. Anzar
Keyword(s):  
Plasma Membrane ◽  
Statistical Analysis ◽  
Sperm Motility ◽  
Propidium Iodide ◽  
Female Reproductive Tract ◽  
Semen Sample ◽  
Membrane Asymmetry ◽  
Progressive Motility ◽  
Bovine Sperm ◽  
Acrosomal Integrity

Extent and timing of alterations in structures and functions of sperm after its placement in the female reproductive tract are important for successful fertilization. To our knowledge, the few reports are available on the kinetics of alterations in bovine sperm structures and functions during pathway to their death. Therefore, the present study was conducted to determine the changes in motility, acrosome and plasma membrane asymmetry in fresh and frozen–thawed semen during incubation at 37°C over the period of 24 h. Semen was collected from 3 breeding beef bulls, pooled, and considered as one replicate (total replicates = 5). Each pooled semen sample was diluted in Tris-citric acid egg yolk glycerol extender (pH 6.8), cooled to +4°C over 90 min, and then cryopreserved by a programmable cell freezer. Fresh (pooled semen) and frozen–thawed semen were incubated at 37°C for 24 h. Each semen sample was evaluated for sperm motility with computer-assisted semen analysis and acrosomal integrity and plasma membrane asymmetry using fluorescein isothiocyanate-peanut agglutinin/propidium iodide and Annexin V/propidium iodide assays, respectively, at 0, 2, 4, 6, 12, and 24 h of incubation at 37°C, with a flow cytometer. Statistical analysis was conducted using PROC MIXED model in statistical analysis system as 2 (semen types) × 6 (times) factorial model, using time as repeated measure. Progressive motility was higher (P < 0.05) in fresh than in frozen–thawed semen until 6 h. Progressive motility declined (P < 0.05) below the threshold level (i.e. 30%) much later (12 h) in fresh as compared with frozen–thawed semen (2 h). However, acrosomal integrity and plasma membrane asymmetry deteriorated (P < 0.05) below threshold at the same time interval (2 h) in both fresh and frozen–thawed semen. Viable sperm (AN–/PI–) remained higher (P < 0.05) during the first 6 h in fresh than in frozen–thawed semen and declined (P < 0.05) below the threshold at 12 h in fresh and at 6 h in frozen–thawed semen. In fresh semen, the necrotic sperm (AN–/PI+) population increased (P < 0.05) over time and reached maximum (97%) at 24 h. In frozen–thawed semen, a mixed population of late apoptotic (53%) and necrotic (34%) sperm was found at 24 h. In conclusion, the alterations in sperm motility, acrosomes, plasma membrane integrity, and asymmetry were slower in fresh than in frozen–thawed semen. Fresh sperm followed necrosis and frozen–thawed sperm underwent necrosis and apoptosis-like pathways, respectively. This study was supported by the Canadian Commonwealth Scholarship Program by the Canadian Bureau for International Education (CBIE), and Agriculture and Agri-Food Canada.

113 Effect of antioxidants on motility and fertility of liquid-stored bovine sperm

2020 ◽  
Vol 32 (2) ◽  
pp. 183
Author(s):  
Y. Honkawa ◽  
T. Fujikawa ◽  
N. Miura ◽  
C. Kubota
Keyword(s):  
Embryo Development ◽  
Sperm Motility ◽  
Sperm Concentration ◽  
Frozen Storage ◽  
Egg Yolk ◽  
P Value ◽  
Motile Sperm ◽  
Progressive Motility ◽  
Bovine Sperm ◽  
Liquid Storage

It is difficult to maintain sperm in liquid storage for a long time, compared with permanent frozen storage in liquid nitrogen. Antioxidants have been reported to improve the quality and fertility of liquid-stored semen. In this study, we investigated whether antioxidants can extend the motility and fertility of frozen-thawed sperm in liquid storage. Frozen-thawed semen from one Japanese black bull (one ejaculate) was diluted in Tris-citrate-fructose (TCF) diluent with 10% (v/v) egg yolk to a sperm concentration of 1×107 spermmL−1. The antioxidants β-mercaptoethanol (βMe) and glutathione (GSH) were added independently, at various concentrations (0.1, 0.5, 1, and 5mM) to sperm suspensions, and these preparations were compared with Control (no added antioxidant). Sperm suspensions were packaged in centrifuge tubes and placed at 17°C in air and monitored daily until sperm motility had stopped (up to 14 days). Sperm motility was analysed by the Sperm Motility Analysis System (SMAS; Ditect Co. Ltd), and the percentage of progressively motile sperm (straight-line velocity (VSL) of &gt;25μm s−1; Grade A classified by WHO manual), compared with that recorded on Day 0 (100%), was determined each day. For evaluation of fertilizing ability, after incubation in liquid storage for 0, 3, 5, and 7 days, sperm were used for IVF with invitro-matured oocytes (30 oocytes per treatment, three replicates). Embryo development was recorded as the proportion of embryos that reached blastocyst by 8 days after IVF. Data for motility were analysed using one-way ANOVA with Tukey test, and embryo development using chi-squared test. A P-value&lt;0.05 was considered statistically significant. At 7 days, the percentage of progressively motile sperm was significantly higher for 0.5, 1, and 5mM βMe than for Control (30.8%, 48.1%, and 50.3%, vs. 0%, respectively). Treatments with 1 and 5mM βMe maintained some sperm progressive motility for 14 days (9.5% and 14.5%). Treatment with GSH showed the same trend at 7 days (32.2%, 36.3%, and 13.7% for 0.5, 1, and 5mM, vs. 0% for Control); 1 and 5mM GSH maintained sperm progressive motility over 10 days (24.8% and 4.4%). In both antioxidant treatments, embryo development was achieved with sperm stored for up to 5 days (Day 0 vs. Day 5 for 0.1mM βMe: 17.6% vs. 13.8%; for 1.0mM GSH: 26.0% vs. 6.7%; for Control: 17.6% vs. 0%). In this study, antioxidants extended both motility and fertility of frozen-thawed bovine sperm in liquid storage. This result suggests the possibility of application to AI using liquid-stored bovine semen.

scholarly journals The Effect of Low-Level Laser Irradiation on Sperm Motility, and Integrity of the Plasma Membrane and Acrosome in Cryopreserved Bovine Sperm

PLoS ONE ◽  
2015 ◽  
Vol 10 (3) ◽  
pp. e0121487 ◽  
Author(s):  
Guilherme Henrique C. Fernandes ◽  
Paulo de Tarso Camillo de Carvalho ◽  
Andrey Jorge Serra ◽  
André Maciel Crespilho ◽  
Jean Pierre Schatzman Peron ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Laser Irradiation ◽  
Sperm Motility ◽  
Low Level ◽  
Low Level Laser ◽  
Bovine Sperm ◽  
Level Laser ◽  
Low Level Laser Irradiation

scholarly journals Sperm preparedness and adaptation to osmotic and pH stressors relate to functional competence of sperm in Bos taurus

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Maharajan Lavanya ◽  
Santhanahalli Siddalingappa Archana ◽  
Divakar Swathi ◽  
Laxman Ramya ◽  
Arunachalam Arangasamy ◽  
...  
Keyword(s):  
Reproductive Tract ◽  
Bos Taurus ◽  
Rejection Rate ◽  
Female Reproductive Tract ◽  
Fertilization Process ◽  
Progressive Motility ◽  
Bovine Sperm ◽  
Functional Attributes ◽  
Adaptive Ability ◽  
Head Area

AbstractThe adaptive ability of sperm in the female reproductive tract micromilieu signifies the successful fertilization process. The study aimed to analyze the preparedness of sperm to the prevailing osmotic and pH stressors in the female reproductive tract. Fresh bovine sperm were incubated in 290 (isosmotic-control), 355 (hyperosmotic-uterus and oviduct), and 420 (hyperosmotic-control) mOsm/kg and each with pH of 6.8 (uterus) and 7.4 (oviduct). During incubation, the changes in sperm functional attributes were studied. Sperm kinematics and head area decreased significantly (p < 0.05) immediately upon exposure to hyperosmotic stress at both pH. Proportion of sperm capacitated (%) in 355 mOsm/kg at 1 and 2 h of incubation were significantly (p < 0.05) higher than those in 290 mOsm media. The magnitude and duration of recovery of sperm progressive motility in 355 mOsm with pH 7.4 was correlated with the ejaculate rejection rate (R2 = 0.7). Using this information, the bulls were divided into good (n = 5) and poor (n = 5) osmo-adapters. The osmo-responsive genes such as NFAT5, HSP90AB1, SLC9C1, ADAM1B and GAPDH were upregulated (p < 0.05) in the sperm of good osmo-adapters. The study suggests that sperm are prepared for the osmotic and pH challenges in the female reproductive tract and the osmoadaptive ability is associated with ejaculate quality in bulls.

scholarly journals Effects of Media and Frame Rates on Hyperactivated Bovine Sperm Motility and an Analysis of Sperm Motility Subpopulation Structures in Sex-Sorted and Non-Sorted Semen

2021 ◽  
Author(s):  
Chihiro Kanno ◽  
Sun-Sik Kang ◽  
Kentaro Q. Sakamoto ◽  
Yojiro Yanagawa ◽  
Seiji Katagiri ◽  
...  
Keyword(s):  
Cluster Analysis ◽  
Discriminant Analysis ◽  
Sperm Motility ◽  
Computer Assisted ◽  
Motile Sperm ◽  
Progressive Motility ◽  
Bovine Sperm ◽  
Using Data ◽  
Effects Of Media ◽  
Oviductal Fluid

We attempted to establish an objective method to accurately evaluate the motility of bull sperm and examined the effects of media for sperm suspensions and frame rates on data of computer-assisted sperm motility analysis (CASA). Sperm incubated in Brackett and Oliphant medium (BO) more clearly showed hyperactivation-like motility than those in synthetic oviductal fluid. Sperm images captured at 150 frames per second (fps) showed a trajectory that was closer to the real pathway than those at other frame rates (30, 50, and 75 fps). We then examined the characteristics of sex-sorted and non-sorted semen using a cluster analysis followed by a discriminant analysis of sperm motility in BO at 150 fps. The results indicated that sex-sorted semen contained sperm with hyperactivation-like motility as the main subpopulation immediately after thawing and this subpopulation decreased after 2-h incubation. The main subpopulation in non-sorted semen had progressive motility that was maintained during incubation. In conclusion, usage of BO for sperm suspensions and capturing sperm motility at 150 fps by CASA were appropriate for evaluating bovine sperm motility. A discriminant analysis using data from a cluster analysis of motile sperm has the ability to accurately describe differences in the structures of sperm motility subpopulations.

scholarly journals Effect of warming temperatures on donkey sperm vitrification in 0.5 mL straws in comparison to conventional freezing

2019 ◽  
Vol 17 (3) ◽  
pp. e0406 ◽  
Author(s):  
Maria Diaz-Jimenez ◽  
Jesus Dorado ◽  
Cesar Consuegra ◽  
Blasa Pereira ◽  
Isabel Ortiz ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Slow Freezing ◽  
Kinematic Parameters ◽  
Plasma Membrane Integrity ◽  
Progressive Motility ◽  
Acrosome Integrity ◽  
Sperm Freezing ◽  
Conventional Freezing

Aim of study: There is little information about vitrification of sperm in large volumes (up to 0.5 mL). This study aimed to develop the vitrification technique in 0.5 mL straws in donkey sperm, evaluating the effect of three warming temperatures.Area of study: Cordoba, Spain.Material and methods: Ejaculates from five donkeys were divided in four groups: one control subjected to conventional slow freezing (C) and three vitrified in 0.5 mL straws and warmed using different protocols (W1: 37ºC/30s, W2: 43ºC/20s and W3: 70ºC/8s+37ºC/52s). Sperm motility, kinematic parameters, plasma membrane and acrosome integrity were evaluated. Conventional freezing resulted in significantly higher values for total (42.7±19.6%), and progressive motility (30.3±16.7%), plasma membrane (49.1±10.4%) and acrosome integrity (39.6±14.5%) respect to vitrification method.Main results: Values after warming ranged between 0.2-2.8% for total motility; 0.2-2.1% for progressive motility; 5.5-20.0% for plasma membrane integrity and 14.5-29.8% for acrosome integrity in all warming protocols after sperm vitrification. However, no differences were found between W3 and C for kinematic parameters; and W3 resulted in significantly higher values for membrane integrity (20.0±11.0%) in comparison to W1 (5.5±3.6%) and W2 (9.3±8.4%).Research highlights: High warming rates seem to be better for donkey sperm vitrification in large volumes; but this methodology is still not an alternative to conventional sperm freezing.

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 Influence of Seasons and Extenders on Quality and Freezability of Gir Bull Semen under Middle Gujarat Climate

2018 ◽  
Vol 13 (03) ◽  
Author(s):  
D. V. Chaudhari ◽  
J. A. Patel ◽  
K. K. Hadiya ◽  
A. J. Dhami
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Winter Season ◽  
Egg Yolk ◽  
Quality Parameters ◽  
Summer Season ◽  
Progressive Motility ◽  
Bull Semen

            The study was conducted to evaluate the seasonal influence (peak winter and summer) and the efficacy of three extenders (egg yolk based TFYG extender and egg yolk free soya bean based commercial extenders Optixcell and Andromed) on quality and freezability of Gir bull semen in Middle Gujarat. Semen ejaculates (6/bull/season, total36) revealed mean ejaculate volume 6.49±0.30 ml, sperm concentration1212.36±58.10 million/ml, progressive motility 74.17±0.78 %, live sperm 81.39±0.80 %, abnormal sperm 7.36±0.31 %, and sperm with intact plasma membrane 81.31±0.98 % and intact acrosome 94.81±0.24 %. Only the progressive sperm motility was significantly (P<0.05) higher(76.39±0.97 % vs. 71.94±1.00 %) with lesser sperm abnormality(6.17±0.37 % vs. 8.56±0.30 %) during winter than in summer. Semen samples split diluted with TFYG, Optixcell and Andromed extenders recorded the overall mean values of progressive sperm motility, livability, abnormality, plasma membrane integrity and acrosomal integrity during winter season as 77.87±0.51, 77.50±0.45, 5.56±0.20,76.02±0.81 and 94.35±0.29 on dilution; 72.41±0.51, 70.50±0.64, 5.96±0.26, 71.20±0.79 and 93.09±0.32 at pre-freeze stage; 41.30 ±0.94, 50.28±1.03, 9.15±0.31, 29.89±0.40 and 90.65±0.40 at post-thaw stage, respectively. The respective values in summer season were 72.13±0.60, 75.50±0.60, 7.48±0.25, 75.61 ±0.55 and 94.09±0.30 on dilution; 65.46±0.66, 69.41±1.05, 8.89±0.28, 69.70±0.66 and 92.63 ±0.33 at pre-freeze stage; 31.48±0.52, 45.09±0.85, 13.48±0.33, 26.85±0.71 and91.26±0.38 at post-thaw stage.  The overall mean sperm post-thaw motility/longevity at 0, 30, 60 and 120 min of incubation at 37°C was 41.20±1.51, 35.19±1.47, 28.80±1.75 and 17.50±1.47 % during winter season and 31.57±0.89, 26.20±0.77, 20.37±0.83 and 13.80±0.77% in summer season, respectively. The initial quality as well as freezability of semen in terms of motile, live, normal and HOS reactive sperm including post thaw longevity were better in winter season than in summer season. Further, the values of all the five semen quality parameters studied were comparatively better in Optixcell than TFYG and Andromed extenders with significant differences only in sperm progressive motility in both the seasons.The season x extender interaction was not significant for any of the sperm quality parameters studied.

Cryopreservation of Indian Red Jungle Fowl (Gallus Gallus Murghi) Spermatozoa: A First Approach

2017 ◽  
Vol 10 (1) ◽  
pp. 6-11 ◽  
Author(s):  
Bushra Allah Rakha ◽  
Muhammad Sajjad Ansari ◽  
Iftikhar Hussain ◽  
Maqsood Anwar ◽  
Shamim Akhter ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Gallus Gallus ◽  
Freeze Thaw ◽  
Red Jungle Fowl ◽  
Frozen Semen ◽  
Sperm Plasma Membrane ◽  
Acrosomal Integrity

The experiment was designed to show the capacity of Indian Red Jungle Fowl ( Gallus gallus murghi) sperm to be cryopreserved. The changes in characteristics of Indian Red Jungle Fowl spermatozoa during different phases of cryopreservation with the glycerol cryoprotectant and straw packaging were recorded. Semen from eight mature cocks were pooled, diluted at 37 °C in a Tselutin poultry extender, cooled to 4 °C in 2 h (0.275 min−1) and equilibrated for 10 min after the addition of 11% glycerol. Cooled semen was filled in 0.5 mL French straws, kept over liquid nitrogen vapour for 10 min and plunged into liquid nitrogen. Frozen semen was thawed at 37 °C for 30 s. Sperm motility, plasma membrane plasticity, livability, and acrosomal integrity were assessed at the stage of pre-dilution (37 °C), post-dilution, cooling, equilibration and freeze-thawing. The experiment was repeated five times. Sperm motility did not change at post-dilution and cooling and averaged 83.3±3.1%. However, motility declined ( P<0.05) to 66.0±2.4% and 45.0±6.5% at post-equilibration and thawing stages, respectively. The sperm plasma membrane plasticity at the pre-dilution stage was 74.8±1.4% and decreased ( P<0.05) to 62.3±1.7%, 51.7±1.7%, and 42.0±1.2% at the stages of post-dilution, cooling and freeze-thawing, respectively. A decline ( P < 0.05) in acrosomal integrity was observed at post-cooling (83.8±2.4%), equilibration (80.8±0.8%) and thawing (73.8±2.4%). It is concluded that Indian Red Jungle sperm seem to adapt quite well to the freeze-thaw process that mainly alters the motility capacities and much less the acrosomal integrity. To our knowledge, this is the first ever report on cryopreservation of Indian Red Jungle Fowl spermatozoa.

scholarly journals Use of computer-assisted sperm motility assessment and multivariate pattern analysis to characterize ejaculate quality in Mohor gazelles (Gazella dama mhorr): effects of body weight, electroejaculation technique and short-term semen storage

Reproduction ◽  
2001 ◽  
pp. 265-273 ◽  
Author(s):  
T Abaigar ◽  
M Cano ◽  
AR Pickard ◽  
WV Holt
Keyword(s):  
Body Weight ◽  
Plasma Membrane ◽  
Sperm Motility ◽  
Pattern Analysis ◽  
Semen Quality ◽  
Computer Assisted ◽  
Short Term ◽  
Non Linear ◽  
Maximum Voltage ◽  
Acrosomal Integrity

Subjective and objective semen assessments were performed on 18 male Mohor gazelles (Gazella dama mhorr). Sperm motility assessments combined with sperm plasma membrane and acrosomal integrity evaluations were undertaken as part of a captive breeding programme. The primary objective was to test methodology for short-term preservation of gazelle semen for artificial insemination (storage in N-[Tris(hydroxymethyl)methyl]-2-aminoethanesulphonic acid-Tris diluent (TEST) for up to 96 h at 17 degrees C). However, the secondary objective was to investigate phenotypic and genotypic influences on semen quality within this small population, which was established in 1971 with only 12 genetic founders. Sperm motility was measured by computer-assisted semen assessment and the data were analysed using a pattern analysis technique to detect and quantify naturally occurring sperm subpopulations within the semen samples. Four sperm subpopulations distinguishable by their motion characteristics were detected. The relative frequencies of two subpopulations (population 2: highly motile, non-linear; and population 4: poorly motile, non-linear) in fresh semen were correlated with the maximum voltage used during electroejaculation. The frequency of subpopulation 2 was negatively correlated with maximum voltage (r = -0.875, P < 0.0001) and the frequency of subpopulation 4 was positively correlated (r = 0.727, P < 0.005). The frequencies of all subpopulations varied significantly among the animals sampled (chi-squared = 2577.6, degrees of freedom = 54, P < 0.0001) and subpopulation 4 was also correlated with body weight (r = -0.59, P < 0.005). Semen stored at 17 degrees C retained motility, plasma membrane and acrosomal integrity for 48 h, but these measures decreased thereafter. The frequency of a sperm subpopulation showing uncoordinated but active motility increased significantly over the first 48 h and then decreased.

Arrangement of Mitochondria in Spermatozoa of the Mexican Axolotl, Ambystoma Mexicanum

1973 ◽  
Vol 31 ◽  
pp. 626-627
Author(s):  
Ezzatollah Keyhani ◽  
Larry F. Lemanski ◽  
Sharon L. Lemanski
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Substrate Oxidation ◽  
Ambystoma Mexicanum ◽  
Axial Filament ◽  
Mexican Axolotl ◽  
Middle Piece

Energy for sperm motility is provided by both glycolytic and respiratory pathways. Mitochondria are involved in the latter pathway and conserve energy of substrate oxidation by coupling to phosphorylation. During spermatogenesis, the mitochondria undergo extensive transformation which in many species leads to the formation of a nebemkem. The nebemkem subsequently forms into a helix around the axial filament complex in the middle piece of spermatozoa.Immature spermatozoa of axolotls contain numerous small spherical mitochondria which are randomly distributed throughout the cytoplasm (Fig. 1). As maturation progresses, the mitochondria appear to migrate to the middle piece region where they become tightly packed to form a crystalline-like sheath. The cytoplasm in this region is no longer abundant (Fig. 2) and the plasma membrane is now closely apposed to the outside of the mitochondrial layer.

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