Relationships among in vivo fertility, computer-analysed motility and in vitro Ca2+ flux in bovine spermatozoa

1994 ◽  
Vol 74 (1) ◽  
pp. 53-58 ◽  
Author(s):  
J. L. Bailey ◽  
M. M. Buhr ◽  
L. Robertson
Keyword(s):  
Rate Of Change ◽  
Calcium Flux ◽  
Computer Assisted ◽  
Spermatozoa Motility ◽  
Head Displacement ◽  
Intracellular Ca ◽  
Bovine Spermatozoa ◽  
Lateral Head

Correlations among computer-assisted spermatozoa motility analyses, Ca2+ fluxes and in vivo fertility of bovine spermatozoa based on a total of 4482 inseminations were investigated in each of four ejaculates from six bulls. The Ca2+ parameters assessed the rate of change in intra- and extracellular Ca + in fresh and cryopreserved spermatozoa from the same ejaculates and were described in another study. Of the seven motility parameters of cryopreserved semen investigated, all differed significantly among bulls but none were related to the in vivo fertility of cryopreserved semen. The amplitude of lateral head displacement, a motility parameter associated with hyperactivation, was positively correlated with the intracellular Ca2+ levels and the rate of Ca2+ accumulation of cyropreserved spermatozoa. The highest fertility was observed when initial extracellular Ca2+ for cryopreserved spermatozoa was high and when Ca2+ efflux rates of cryopreserved cells approached the higher efflux rates of fresh spermatozoa. Fertility was reduced when cryopreserved spermatozoa had initial internal Ca2+ levels greater than those of fresh spermatozoa or when cryopreserved spermatozoa internalized Ca2+ rapidly. Calcium flux, but not motility, may predict fertilizing capacity of cryopreserved bovine spermatozoa. Key words: Calcium, computer-analysed motility, fertility, spermatozoa, bull

scholarly journals Effect of three commercial extenders on sperm motility and fertility in liquid ram semen stored at 15 °C or 5 °C

2019 ◽  
Vol 67 (3) ◽  
pp. 430-444
Author(s):  
Ander Arando ◽  
Juan Vicente Delgado ◽  
José Manuel León ◽  
Sergio Nogales ◽  
Francisco Javier Navas-González ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Soybean Lecithin ◽  
Egg Yolk ◽  
Kinematic Parameters ◽  
Progressive Motility ◽  
Head Displacement ◽  
Liquid Storage ◽  
Lateral Head

The effect of different extenders on sperm motility and fertility was evaluated during liquid storage of ram semen at 5 °C and 15 °C. The semen was collected, pooled and diluted in three commercial extenders: Inra 96® (INRA) based on skimmed milk, Biladyl® A fraction (BIL) based on egg yolk, and Ovixcell® (OVIX) based on soybean lecithin. Then, sperm motility was evaluated at 0, 6, 24, 48, 72 and 96 h. In order to evaluate fertility, samples stored at 15 °C were used after dilution in INRA and OVIX. Results showed that progressive motility was significantly higher up to 72 h of storage in sperm samples maintained at 5 °C in comparison with 15 °C, similarly for each tested diluent. When samples were stored at 5 °C in OVIX, kinematic parameters such as velocity (except curvilinear velocity, VCL), trajectory [linearity (LIN), straightness (STR), wobble (WOB)], amplitude of lateral head displacement (ALH) and beat/cross frequency (BCF) were higher than in INRA and BIL. No significant differences in pregnancy rate were detected between INRA (62.6%) and OVIX (58.9%). In conclusion, liquid storage at 5 °C with OVIX extender is an interesting option since non-animal components are used, and this extender offers similar in vitro and in vivo efficacy as other extenders containing animal components.

scholarly journals IN VITRO EFFECTS OF ISOQUERCITRIN ON SELECTED VITALITY MARKERS OF BOVINE SPERMATOZOA

2019 ◽  
Vol 1 (2) ◽  
pp. 18-23
Author(s):  
Filip Benko ◽  
Hana Greifová ◽  
Eva Tvrdá
Keyword(s):  
Physiological Saline ◽  
Biological Properties ◽  
Computer Assisted ◽  
Movement Activity ◽  
Spermatozoa Motility ◽  
Physiological Saline Solution ◽  
Bovine Spermatozoa ◽  
In Vitro Effects

The aim of this study was to evaluate the dose- and time-dependent in vitroeffects of isoquercitrin (ISO), a natural flavonoid with numerous biological properties on bovine spermatozoa during three different time periods (0 h, 2 h, 24 h). Bovine semen samples were diluted and cultivated in physiological saline solution containing 0.5% DMSO together with 200, 100, 50, 10, 5 and 1 μmol/L ISO. Spermatozoa motility was measured using the HTM IVOS CASA (Computer Assisted Semen Analyzer) system. The viability of spermatozoa was assessed by the metabolic (MTT) assay, production of superoxide radicals was quantified using the nitroblue tetrazolium (NBT) test, and chemiluminescence was used to evaluate the generation of reactive oxygen species (ROS). The results of the movement activity showed a significant increase in the motility during long term cultivation in case of concentrationsranging between 5 and 50 μmol/L ISO (P<0.05; 24 h). At the same time, supplementation of several concentrations of ISO led to a significant preservation of the cell viability (P<0.05 in the case of 50 μmol/L, P<0.01 with respect to 1 and 5 μmol/L, and P<0.001 in relation to 10 μmol/L; 24 h). ISO addition at 10 and 50 μmol/L also provided a significantly higher protection against superoxide (P<0.05) and ROS (P<0.001) overgeneration after a 24 h cultivation. We may suggest that supplementation of ISO to bovine spermatozoa, particularly at concentrations ranging between 10 and 50 μmol/L, may offer protection to the motility, viability and oxidative status of the sperm cells, particularly notable at 24 h.

371 DOES Percoll™ GRADIENT CENTRIFUGATION CAUSE HYPERACTIVATION ON CRYOPRESERVED BOVINE SPERMATOZOA?

2010 ◽  
Vol 22 (1) ◽  
pp. 342
Author(s):  
L. Z. Oliveira ◽  
R. P. Arruda ◽  
E. C. C. Celeghini ◽  
A. F. C. Andrade ◽  
A. C. Lucio ◽  
...  
Keyword(s):  
Density Gradient ◽  
Calcium Influx ◽  
Ph Monitoring ◽  
Semen Analysis ◽  
Gradient Centrifugation ◽  
Control Group ◽  
Computer Assisted ◽  
Head Displacement ◽  
Bovine Spermatozoa

The density difference of 0.06% between X- and Y-bearing bovine spermatozoa has the potential to provide for separation X-bearing spermatozoa on specific gradient solutions of Percoll™ (Hossepian de Lima VFM et al. PCT/BR2004/000009). The question remains whether Percoll™ density gradient centrifugation induces the hyperactivation of spermatozoa, because it is believed that some decapacitating proteins are removed from the sperm surface during the process. Thus, the objective of this study was to evaluate if Percoll™ centrifugation causes hyperactivation of cryopreserved bovine spermatozoa. Semen doses were collected from six bulls of different breeds, including three taurine and three Zebu animals, and four ejaculates per bull were evaluated. The semen samples were thawed and evaluated before (control) and after (sexed group) centrifugation (500 g) in discontinuous Percoll™ density gradient (Hossepian Lima VFM et al. PCT/BR2004/000009). Sperm motility was assessed by Computer-Assisted Semen Analysis (CASA, Hamilton Thorne Biosciences, Beverly, MA, USA) and the results (mean ± SEM) were submitted to ANOVA. Higher (P < 0.0001) total and progressive sperm motilities were observed in sexed (85.0 ± 1.8 and 75.9 ± 1.7%, respectively) than in the control group (69.9 ± 1.7 and 59.2 ± 1.6%, respectively). The average path velocity, straight-line velocity and curvilinear velocity (VCL) were lower (P < 0.01) in sexed (91.3 ± 1.1, 79.8 ± 0.8, and 137.6 ± 3.5 (im/s, respectively) than in control group (107.6 ± 4.8, 91.1 ± 4.1, and 173.2 ± 8.9 (im/s, respectively). The amplitude of lateral head displacement (ALH) was higher (P < 0.0001) in control (6.7 ± 0.3 μm) than in sexed semen (5.1 ± 0.2 μm) and beat cross frequency was higher (P < 0.05) in sexed (35.9 ± 0.9 Hz) than in control semen (33.1 ± 1.0 Hz). The straightness and linearity (LIN) were higher (P < 0.01) in sexed (87.4 ± 0.7 and 61.4 ± 1.4%, respectively) than in control semen (84.4 ± 0.7 and 55.5 ± 1.2%, respectively). Regarding the results obtained in the present study, it was not possible to infer that the method used induced sperm hyperactivation. According to Marquez and Suarez (2007 Biol. Reprod., 76, 660-665), an increase in pH and intraflagellar calcium plays a key role in the axoneme changes, promoting an increase in ALH (from 9 to 13 μm) and VCL (from 200 to 315 μm/s), as well as a decrease in LIN (from 57 to 25%), which characterizes the presence of hyperactivated cells in the sample. In the present study, centrifuged spermatozoa presented significantly lower ALH and VCL and higher LIN compared to control semen samples. Therefore, the presence of glucose in the Percoll™ stock solution and constant pH monitoring could have prevented alkalinization of the medium, thereby preventing the occurrence of intraflagelar calcium influx and the consequent triggering of in vitro sperm hyperactivation.

Computer assisted sperm analysis: Relationship between the movement characteristics of buffalo spermatozoa and sire fertility

2016 ◽  
Author(s):  
Raushan K Singh ◽  
A. Kumaresan ◽  
M. A. Mir ◽  
P. Kumar ◽  
S. Chhillar ◽  
...  
Keyword(s):  
Computer Assisted ◽  
Progeny Testing ◽  
Kinematic Parameters ◽  
Murrah Buffalo ◽  
Sperm Analysis ◽  
Head Displacement ◽  
Movement Characteristics ◽  
Frozen Semen ◽  
Lateral Head

The present study was undertaken to identify the differences in sperm kinematics between buffalo bulls with different fertility ratings. Murrah buffalo bulls (n=9) that were routinely used for breeding purpose under progeny testing programme were utilized for the study. Bull fertility was determined based on in vivo fertility trials and the conception rates (CR) were adjusted for different non-genetic parameters. Based on the adjusted CR, bulls were classified into high, medium and low fertile group. Frozen semen samples of these bulls were obtained and sperm kinematic parameters were assessed using a computer assisted sperm analyzer. The kinematic parameters analyzed included the curvilinear velocity (VCL), the linear velocity (VSL), the average path velocity (VAP), the amplitude of lateral head displacement (ALH), the linearity (LIN), the straightness coefficient (STR) and the beat cross frequency (BCF). In high fertile bulls, the proportion of motile spermatozoa was higher (p<0.001) than the medium and low fertile bulls. The VAP and VCL of sperm motion were significantly higher (P<0.05) in high fertile bulls compared to either medium or low fertile bulls. The VSL was significantly lower in low fertile bulls (P<0.005) compared to either high or medium fertile bulls. Spermatozoa from high fertile bulls had significantly higher (P<0.05) BCF, STR, ALH and LIN compared to either medium or low fertile bulls. Buffalo bull fertility was significantly and positively correlated with sperm motility, VAP, VSL, VCL and ALH.

scholarly journals In vitro effect of ferrous sulphate on bovine spermatozoa motility parameters, viability and Annexin V-labeled membrane changes

PLoS ONE ◽  
2021 ◽  
Vol 16 (9) ◽  
pp. e0257766
Author(s):  
Zuzana Knazicka ◽  
Hana Duranova ◽  
Veronika Fialkova ◽  
Michal Miskeje ◽  
Tomas Jambor ◽  
...  
Keyword(s):  
Cell Viability ◽  
Ferrous Sulphate ◽  
Annexin V ◽  
Computer Assisted ◽  
Spermatozoa Motility ◽  
Bovine Spermatozoa ◽  
High Concentrations ◽  
Vitro Effect ◽  
Motility Parameters

The aim of this study was to assess the dose- and time-dependent in vitro effects of ferrous sulphate (FeSO4.7H2O) on the motility parameters, viability, structural and functional activity of bovine spermatozoa. Spermatozoa motility parameters were determined after exposure to concentrations (3.90, 7.80, 15.60, 31.20, 62.50, 125, 250, 500 and 1000 μM) of FeSO4.7H2O using the SpermVisionTM CASA (Computer Assisted Semen Analyzer) system in different time periods. Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay, and the Annexin V-Fluos was applied to detect the membrane integrity of spermatozoa. The initial spermatozoa motility showed increased average values at all experimental concentrations compared to the control group (culture medium without FeSO4.7H2O). After 2 h, FeSO4.7H2O stimulated the overall percentage of spermatozoa motility at the concentrations of ≤ 125 μM. However, experimental administration of 250 μM of FeSO4.7H2O significantly (P < 0.001) decreased the spermatozoa motility but had no negative effect on the cell viability (P < 0.05) (Time 2 h). The lowest viability was noted after the addition of ≥ 500 μM of FeSO4.7H2O (P < 0.001). The concentrations of ≤ 62.50 μM of FeSO4.7H2O markedly stimulated (P < 0.001) spermatozoa activity after 24 h of exposure, while at high concentrations of ≥ 500 μM of FeSO4.7H2O the overall percentage of spermatozoa motility was significantly inhibited (P < 0.001) and it elicited cytotoxic action. Fluorescence analysis confirmed that spermatozoa incubated with higher concentrations (≥ 500 μM) of FeSO4.7H2O displayed apoptotic changes, as detected in head membrane (acrosomal part) and mitochondrial portion of spermatozoa. Moreover, the highest concentration and the longest time of exposure (1000 μM of FeSO4.7H2O; Time 6 h) induced even necrotic alterations to spermatozoa. These results suggest that high concentrations of FeSO4.7H2O are able to induce toxic effects on the structure and function of spermatozoa, while low concentrations may have the positive effect on the fertilization potential of spermatozoa.

scholarly journals Changes in flagellar bending during the course of hyperactivation in hamster spermatozoa

Reproduction ◽  
2003 ◽  
pp. 43-51 ◽  
Author(s):  
M Kinukawa ◽  
M Nagata ◽  
F Aoki
Keyword(s):  
Female Genital Tract ◽  
Movement Pattern ◽  
Rate Of Change ◽  
Motility Pattern ◽  
Head Displacement ◽  
Bend Angles ◽  
Lateral Head ◽  
Sperm Movement ◽  
Mammalian Spermatozoa

The motility pattern of mammalian spermatozoa changes during migration in the female genital tract and during incubation in vitro. This change in motility is termed hyperactivation. Hyperactivated spermatozoa swim vigorously in 'whiplash', 'figure-8' or 'small circle' trajectories. In this study, a quantitative analysis was carried out of the changes in the motility pattern of hamster spermatozoa during incubation to investigate the mechanism regulating hyperactivation. In the culture system used in this study, hyperactivation occurred 4 h after incubation. Several parameters in the analysis of sperm movement pattern were examined. Curvilinear velocity, average path velocity and straightness abruptly increased between 2 and 4 h. However, linearity, amplitude of lateral head displacement, beat cross frequency and average wavelength gradually changed with time. In the analysis of flagellar bending, the bend angles were measured after dividing images of the flagellum into short lengths. Flagellar bending changed in different manners in each region during incubation. The asymmetry in the direction of the curve of the head gradually increased with time in the first half of the flagellum. The flexibility, which was determined using the amplitude of bending and the rate of change in bend angles, abruptly decreased between 10 min and 1 h, and then increased between 2 and 4 h in the first half of flagellum. These results indicate that complex physiological changes occur before hyperactivation.

In vitroeffect of various cryoprotectants on the semen quality of endangered Oravka chicken

Zygote ◽  
2017 ◽  
Vol 26 (1) ◽  
pp. 33-39 ◽  
Author(s):  
Andrea Svoradová ◽  
Lenka Kuželová ◽  
Jaromír Vašíček ◽  
Andrej Baláži ◽  
Emília Hanusová ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Semen Quality ◽  
Semen Analysis ◽  
Lower Percentage ◽  
Computer Assisted ◽  
Spermatozoa Motility ◽  
Progressive Movement ◽  
Animal Genetic Resources

SummaryWe aimed to compare the effect of three different permeating cryoprotectants on the post-thaw spermatozoa quality. Pooled semen from Oravka cock line (n= 6) was diluted inKobidil+extender and frozen in cryoprotectant solutions containing 8% dimethylsulfoxide (DMSO), 8% ethylene glycol (EG) or 8% glycerol (GL) in liquid nitrogen vapours before being plunged into the liquid nitrogen. Spermatozoa motility parameters were assessedin vitroafter freezing–thawing by a computer-assisted semen analysis (CASA) system and viability status was examined using fluorescent probes. The lower percentage (P <0.05) of motile and progressively moving spermatozoa immediately after thawing were obtained in all experimental groups (DMSO, EG, GL) compared with the control. Significant (P< 0.05) differences in total motility and progressive movement between GL and DMSO, EG groups were observed. However, the higher number (P <0.05) of acrosome damaged spermatozoa was found in the DMSO and EG groups and no significant differences were observed in the GL group compared with the control. Differences (P< 0.05) between experimental groups and the control in the results of spermatozoa necrosis were observed. No significant differences in the percentage of apoptotic spermatozoa were found between control and experimental groups. However, significant differences (P< 0.05) in number of live and necrotic spermatozoa between GL and DMSO, EG groups were examined. The findings of the present study indicate that glycerol seems to be suitable for semen cryopreservation in the gene banks. In addition, fertility evaluationin vivois needed in order to evaluate the possible contribution for the bank of animal genetic resources.

Cardiovascular Effects of Micromeria graeca (L.) Benth. ex Rchb in Normotensive and Hypertensive Rats

2020 ◽  
Vol 20 (8) ◽  
pp. 1253-1261
Author(s):  
Mourad Akdad ◽  
Mohamed Eddouks
Keyword(s):  
Blood Pressure ◽  
Cardiovascular System ◽  
Arterial Blood Pressure ◽  
Antihypertensive Activity ◽  
Computer Assisted ◽  
Hypertensive Rats ◽  
Vasorelaxant Effect ◽  
Arterial Blood

Aims: The present study was performed in order to analyze the antihypertensive activity of Micromeria graeca (L.) Benth. ex Rchb. Background: Micromeria graeca (L.) Benth. ex Rchb is an aromatic and medicinal plant belonging to the Lamiaceae family. This herb is used to treat various pathologies such as cardiovascular disorders. Meanwhile, its pharmacological effects on the cardiovascular system have not been studied. Objective: The present study aimed to evaluate the effect of aqueous extract of aerial parts of Micromeria graeca (AEMG) on the cardiovascular system in normotensive and hypertensive rats. Methods: In this study, the cardiovascular effect of AEMG was evaluated using in vivo and in vitro investigations. In order to assess the acute effect of AEMG on the cardiovascular system, anesthetized L-NAME-hypertensive and normotensive rats received AEMG (100 mg/kg) orally and arterial blood pressure parameters were monitored during six hours. In the sub-chronic study, rats were orally treated for one week, followed by blood pressure assessment during one week of treatment. Blood pressure was measured using a tail-cuff and a computer-assisted monitoring device. In the second experiment, isolated rat aortic ring pre-contracted with Epinephrine (EP) or KCl was used to assess the vasorelaxant effect of AEMG. Results: Oral administration of AEMG (100 mg/kg) provoked a decrease of arterial blood pressure parameters in hypertensive rats. In addition, AEMG induced a vasorelaxant effect in thoracic aortic rings pre-contracted with EP (10 μM) or KCl (80 mM). This effect was attenuated in the presence of propranolol and methylene blue. While in the presence of glibenclamide, L-NAME, nifedipine or Indomethacin, the vasorelaxant effect was not affected. Conclusion: This study showed that Micromeria graeca possesses a potent antihypertensive effect and relaxes the vascular smooth muscle through β-adrenergic and cGMP pathways.

Paradoxical effect of salbutamol in a model of acute organophosphates intoxication in guinea pigs: role of substance P release

2007 ◽  
Vol 292 (4) ◽  
pp. L915-L923 ◽  
Author(s):  
Jaime Chávez ◽  
Patricia Segura ◽  
Mario H. Vargas ◽  
José Luis Arreola ◽  
Edgar Flores-Soto ◽  
...  
Keyword(s):  
Substance P ◽  
Guinea Pigs ◽  
Smooth Muscle Contraction ◽  
In Vivo Experiments ◽  
Intracellular Ca ◽  
Neurokinin 1 ◽  
Substance P Release

Organophosphates induce bronchoobstruction in guinea pigs, and salbutamol only transiently reverses this effect, suggesting that it triggers additional obstructive mechanisms. To further explore this phenomenon, in vivo (barometric plethysmography) and in vitro (organ baths, including ACh and substance P concentration measurement by HPLC and immunoassay, respectively; intracellular Ca2+ measurement in single myocytes) experiments were performed. In in vivo experiments, parathion caused a progressive bronchoobstruction until a plateau was reached. Administration of salbutamol during this plateau decreased bronchoobstruction up to 22% in the first 5 min, but thereafter airway obstruction rose again as to reach the same intensity as before salbutamol. Aminophylline caused a sustained decrement (71%) of the parathion-induced bronchoobstruction. In in vitro studies, paraoxon produced a sustained contraction of tracheal rings, which was fully blocked by atropine but not by TTX, ω-conotoxin (CTX), or epithelium removal. During the paraoxon-induced contraction, salbutamol caused a temporary relaxation of ∼50%, followed by a partial recontraction. This paradoxical recontraction was avoided by the M2- or neurokinin-1 (NK1)-receptor antagonists (methoctramine or AF-DX 116, and L-732138, respectively), accompanied by a long-lasting relaxation. Forskolin caused full relaxation of the paraoxon response. Substance P and, to a lesser extent, ACh released from tracheal rings during 60-min incubation with paraoxon or physostigmine, respectively, were significantly increased when salbutamol was administered in the second half of this period. In myocytes, paraoxon did not produce any change in the intracellular Ca2+ basal levels. Our results suggested that: 1) organophosphates caused smooth muscle contraction by accumulation of ACh released through a TTX- and CTX-resistant mechanism; 2) during such contraction, salbutamol relaxation is functionally antagonized by the stimulation of M2 receptors; and 3) after this transient salbutamol-induced relaxation, a paradoxical contraction ensues due to the subsequent release of substance P.

scholarly journals Effect of taurine on turkey (Meleagris gallopavo) spermatozoa viability and motility

2018 ◽  
Vol 63 (No. 4) ◽  
pp. 127-135 ◽  
Author(s):  
T. Slanina ◽  
M. Miškeje ◽  
F. Tirpák ◽  
M. Błaszczyk ◽  
R. Stawarz ◽  
...  
Keyword(s):  
Meleagris Gallopavo ◽  
Lower Percentage ◽  
Computer Assisted ◽  
Viability Assessment ◽  
Spermatozoa Motility ◽  
In Vitro Incubation ◽  
Time Periods ◽  
Casa System ◽  
Motility Parameters

The effect of taurine on the turkey spermatozoa motility and viability during the in vitro incubation was assessed. Experimental samples were prepared by diluting the raw semen in nine different concentrations of taurine – from 10 mg/ml to 0.078125 mg/ml. The motility parameters were evaluated by the CASA system (Computer Assisted Semen Analyser) using the program Sperm Vision<sup>®</sup> and for spermatozoa viability assessment the eosin-nigrosin staining was performed. Selected parameters were evaluated at six time periods: 0, 1, 2, 3, 4, and 5 h at 5°C and 41°C. At 5°C, a significantly lower percentage of motility and progressive motility was detected only in the samples with the highest concentration of taurine (10 mg/ml) at time 0 and 1. After 2 h of incubation a significant preventive effect of taurine on spermatozoa parameters was observed. The tendency of the taurine effect on motility parameters was different during the in vitro incubation at 41°C. Significantly lower values of motility parameters were detected in all experimental samples in comparison to the control after 5 h. The analysed concentrations of taurine did not significantly affect viability of turkey spermatozoa during all time periods. A higher percentage of dead spermatozoa were observed at 41°C (4.87–9.90%) if compared to 5°C (2.12–4.88%). The results indicated that the addition of taurine (from 2.5 to 7.5 mg/ml) to turkey spermatozoa positively affected the monitored spermatozoa parameters incubated at 5°C.

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