scholarly journals Expression and subcellular localization of the μ-opioid receptor in equine spermatozoa: evidence for its functional role

Reproduction ◽  
2005 ◽  
Vol 129 (1) ◽  
pp. 39-49 ◽  
Author(s):  
Maria Albrizio ◽  
Antonio Ciro Guaricci ◽  
Filippo Maritato ◽  
Raffaele Luigi Sciorsci ◽  
Gaetano Mari ◽  
...  
Keyword(s):  
Opioid Receptor ◽  
Opioid Antagonist ◽  
Computer Assisted ◽  
Sperm Cells ◽  
Exact Nature ◽  
Sperm Analysis ◽  
Head Displacement ◽  
Μ Opioid Receptor ◽  
High Degree ◽  
Lateral Head

The development of fertilizing ability in sperm cells is associated with changes in the plasma membrane. However, to date the exact nature of sequentially activated primary receptors and channels and the signal transduction pathways derived from these remains elusive. We analyzed the expression and localization of the μ-opioid receptossr in equine spermatozoa. A transcript corresponding to the third extracellular loop that selectively binds μ agonists was amplified, sequenced and compared with the known sequences in humans, rats and cattle. The amplification product showed a high degree of nucleotide conservation. By immunofluorescence, μ-opioid receptor labeling was found on the sperm head and on the tail and disappeared in the acrosomal region of acrosome-reacted sperm cells. Immunoblotting revealed two bands of 50 and 65 kDa. Effects of the opioid antagonist naloxone on motility and on viability and capacitation/acrosome reaction were investigated by computer-assisted sperm analysis and Hoechst 33258/chlortetracycline (H258/CTC) staining. Progressive motility was significantly reduced after 3 h incubation in 10−3 M naloxone (P < 0.05), whereas it increased significantly after 5 h in 10−8 M naloxone (P < 0.05). Sperm velocity at 5 h was significantly reduced by the addition of 10−3 M naloxone (P < 0.05), but increased significantly in the presence of 10−8 M (P < 0.001). Curvilinear velocity and amplitude of lateral head displacement in spermatozoa incubated in the presence of naloxone were not indicative of hyperactivation. H258/CTC staining showed that 10−8 M naloxone significantly stimulated capacitation (P < 0.01) after 3 h. However, it had no effect on sperm cell viability and acrosomal status. Overall, this study provides the first evidence that the μ-opioid receptor is expressed in equine spermatozoa and that naloxone significantly affects motility and capacitation.

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 Computer-Assisted Sperm Analysis of Freezable and Nonfreezable Mithun (Bos frontalis) Semen

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
P. Perumal ◽  
S. K. Srivastava ◽  
S. K. Ghosh ◽  
K. K. Baruah
Keyword(s):  
Objective Analysis ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Progressive Motility ◽  
Head Displacement ◽  
Straight Line ◽  
Computer Assisted Sperm Analysis ◽  
Lateral Head ◽  
Motility Parameters

The present study was undertaken to assess the motility and velocity parameters of sperm of freezable and nonfreezable ejaculates by computer-assisted sperm analyser (CASA) such as Hamilton-Thorne Semen Analyser IVOS 11 in mithun semen. Fifty ejaculates (twenty-five ejaculates each for freezable and nonfreezable semen ejaculates) were collected from ten matured mithun bulls. CASA parameters, motility parameters such as forward progressive motility (FPM) (%), nonprogressive motility (NPM) (%), total motility (TM) (%), and static sperms (SM) (%); velocity parameters such as curvilinear velocity (VCL) (μm/sec), straight line velocity (VSL) (μm/sec), average path velocity (VAP) (μm/sec), linearity (LIN) (%), straightness (STR) (%), wobble (WOB) (%), amplitude of lateral head displacement (ALH) (μm), and beat/cross-frequency (BCF) (Hz) were measured by CASA analyser. The result revealed that these parameters varied significantly (P<0.05) between the freezable and nonfreezable ejaculates and freezable ejaculates have significantly (P<0.05) higher value than nonfreezable ejaculates. It was concluded that most of the CASA parameters were significantly lower in nonfreezable ejaculates than in freezable ejaculates in mithun and confirmed that the CASA was effective for a quick and objective analysis of motility and velocity parameters in mithun semen.

Computerized human sperm analysis

1994 ◽  
Vol 61 (1) ◽  
pp. 75-80
Author(s):  
G. Bonanni ◽  
A. Colonna ◽  
S. Masiero ◽  
I. Mastrogiacomo
Keyword(s):  
Human Sperm ◽  
Computer Assisted ◽  
Penetration Test ◽  
Sperm Analysis ◽  
Head Displacement ◽  
Seminal Parameters ◽  
Mucus Penetration ◽  
Computer Assisted Sperm Analysis ◽  
Sperm Motion ◽  
Lateral Head

The recent introduction of various devices for computer-assisted sperm analysis allows us to have not only objective measurements of the seminal parameters but also to determine some characteristics of sperm motion such as curvilinear and linear velocity and amplitude of lateral head displacement that cannot be obtained by routine analysis. These new seminal parameters seem to be very useful in the diagnosis of male infertility and, among them, ALH seems to play a more important role; in one of our studies we found a significant correlation between this parameter and the results of the bovine cervical mucus penetration test (Penetrak). Furthermore it is possible to determine sperm subpopulations for each parameter and so it will be possible to express a more accurate judgement not only on the fertilizing capacity of semen but also to know the effects on spermatozoa of different activating and capacitating treatments.

scholarly journals Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and Fields

Animals ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 1993
Author(s):  
Sabrina Gacem ◽  
Jaime Catalán ◽  
Anthony Valverde ◽  
Carles Soler ◽  
Jordi Miró
Keyword(s):  
High Resolution ◽  
Sperm Motility ◽  
Frame Rate ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Head Displacement ◽  
Straight Line ◽  
Counting Chamber ◽  
Computer Assisted Sperm Analysis ◽  
Lateral Head

In order to optimize the donkey sperm motility analysis by the CASA (Computer Assisted Sperm Analysis)-Mot system, twelve ejaculates were collected from six jackasses. Capillary loaded chamber (CLC), ISAS®D4C depths 10 and 20 µm, ISAS®D4C Leja 20 and drop displacement chamber (DDC), Spermtrack® (Spk) depths 10 and 20 µm were used. Sperm kinematic variables were evaluated using each chamber and a high-resolution camera capable of capturing a maximum of 500 frames/second (fps). The optimum frame rate (OFR) (defined according to curvilinear velocity—VCL) was dependent on chamber type. The highest OFR obtained was 278.46 fps by Spk20. Values for VCL, straight-line velocity (VSL), straightness (STR), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) were high in DDC and 10 µm depth. In both DDC 10 and 20 µm, the sperm velocities (VCL, VSL, VAP) and ALH values decreased significantly from the centre to the edges, while Wobble and BCF increased. No defined behavior was observed along the CLC. However, all the kinematic variables had a higher value in a highly concentrated sample, in both chamber types. In conclusion, analyzing a minimum of nine fields at 250 fps from the centre to the edges in Spk10 chamber using a dilution of 30 × 106 sperm/mL offers the best choice for donkey computerised sperm motility analysis.

The effect of chronic treatment with naltrindole, a selective δ-opioid antagonist, on μ-opioid receptor-mediated antinociception in diabetic mice

1993 ◽  
Vol 113 (2) ◽  
pp. 167-171 ◽  
Author(s):  
Junzo Kamei ◽  
Naoya Kawashima ◽  
Yuriko Iwamoto ◽  
Tsutomu Suzuki ◽  
Hiroshi Nagase ◽  
...  
Keyword(s):  
Opioid Receptor ◽  
Chronic Treatment ◽  
Opioid Antagonist ◽  
Diabetic Mice ◽  
Μ Opioid Receptor

Inflammation-reactive astrocytes can be restored with a three drug combination

2014 ◽  
Vol 5 (3) ◽  
pp. 209-209
Author(s):  
E. Hansson ◽  
L. Block ◽  
U. Björklund ◽  
B. Biber
Keyword(s):  
Opioid Receptor ◽  
Primary Cultures ◽  
Reactive Astrocytes ◽  
Opioid Antagonist ◽  
Opioid Agonist ◽  
Cell Parameters ◽  
Ultralow Concentrations ◽  
Inflammatory State ◽  
Μ Opioid Receptor

Abstract Aims In inflammation-reactive astrocytes the cell parameters, Ca2+ signalling, Na+ transporters, cytoskeleton, and release of proinflammatory cytokines are affected. We want to re-establish these parameters with agents, which might have a potential to restore the cells back to a normal non-inflammatory level. Methods Astrocytes in primary cultures were incubated with lipopolysaccharide (LPS) (10 ng/ml) for 24 h to become inflammation-reactive. Different parameters were analysed to verify this inflammation: Ca2+ signalling, Na+/K+-ATPase expression, actin filament organization, and interleukin-1beta release (IL-1β). Results We have used an opioid agonist, endomorphin-1, that stimulates the Gi/o protein of the μ-opioid receptor, an opioid antagonist, naloxone, that inhibits the Gs protein of the μ-opioid receptor in ultralow concentrations, and an anti-epileptic agent, levetiracetam, that counteracts the release of IL-1β. The combination of these three agents managed to activate the Gi/o protein and Na+/K+-ATPase activity, inhibit the Gs protein, and decrease the release of IL-1β. The disorganized actin filaments were restored. Conclusions The findings that the important cell parameters in astrocytes were restored back to their normal non-inflammatory state after the cells were treated with the inflammatory agent LPS could be of clinical significance. It may be useful for the treatment of neuroinflammation and also maybe of long-term pain. The astrocyte networks play a significant role and therefore a well-working intercellular Ca2+ signalling is of utmost importance. Significance These findings put new potential drug regimens towards treatment of neuroinflammation and long-term pain into focus.

90 EFFECT OF EGG YOLK ON THE KINEMATICS AND ACROSOME MEMBRANE INTEGRITY OF COOLED-REWARMED CANINE SPERMATOZOA

2010 ◽  
Vol 22 (1) ◽  
pp. 204
Author(s):  
J. Dorado ◽  
M. J. Galvez ◽  
M. R. Murabito ◽  
S. Demyda ◽  
L. J. De Luca ◽  
...  
Keyword(s):  
Semen Analysis ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Mean Values ◽  
Head Displacement ◽  
Healthy Dogs ◽  
Acrosome Integrity ◽  
Digital Manipulation ◽  
Lateral Head

Tris-egg yolk-based diluents provide adequate cryoprotection for the sperm of most species. This study was conducted to compare the ability of Tris-glucose extender containing 2 different concentrations of egg yolk to maintain sperm motility and acrosome integrity of canine spermatozoa during 72 h of preservation. For this purpose, a total of 20 ejaculates from 4 clinically healthy dogs (2 Spanish Greyhound, 1 German Pointer, and 1 Crossbreed) were collected by digital manipulation. The sperm-rich fraction of each ejaculate was divided into 2 aliquots. Then, they were diluted in Tris-based extender and centrifuged at 700g for 8 min. Sperm pellets were resuspended in either Tris buffer added to 20% (EY20) or 10% centrifuged egg yolk (EY10) and cooled to 5°C over 72 h. The effects of these extenders on motility and acrosome integrity were assessed objectively using a computer-aided semen analyzer (Sperm Class Analyzer, Microptic SL, Spain) and Spermac® staining, respectively. Each cooled-rewarmed semen sample was evaluated after 24, 48, and 72 h of preservation. Sperm motion parameters shown by computer-assisted semen analysis (CASA) are progressively motile (PMS) and motile spermatozoa (MS), curvilinear velocity (CLV), average path velocity (APV), progressive speed (SLV), and lateral head displacement (LHD). Data were statistically analysed by ANOVA. Dependent variables expressed as percentages were arsine-transformed before analysis. Differences between mean values were evaluated by the Duncan method. Data were presented as mean ± SEM. Differences were considered significant when P < 0.05. Analyses were performed using the statistical package SPSS 12.0. A total of 98 172 motile sperm trajectories were analyzed by CASA: 52 259 in EY20 and 45 913 in EY10. After 24, 48, and 72 h of preservation, MS and PMS were statistically higher (P < 0.01) in EY20. No significant differences were found for LHD using either extender over a 72-h period. No significant differences were observed for CLV using either extender during the first 2 days. At Day 3, CLV data were significantly higher (P < 0.01) in EY20. Similarly, from Day 2, APV was significantly higher (P < 0.001) in EY20. After 24 h of preservation, SLV was statistically higher (P < 0.001) in EY10, whereas the opposite tendency was found at Day 3. No significant differences were observed for SLV using either extender after 48 h of preservation. During the first 2 days, acrosome integrity was statistically higher (P < 0.001) in EY20. At hour 72, higher acrosome integrity (P < 0.001) was observed in EY10. In conclusion, we have observed that the EY20 extender provided higher motility after 72 h of chilled preservation; however, the acrosome membrane integrity was better preserved in EY10.

17 Increased inbreeding levels negatively affect sperm kinetics and motility in Purebred Spanish horses

2021 ◽  
Vol 33 (2) ◽  
pp. 116
Author(s):  
Y. Pirosanto ◽  
A. Molina ◽  
M. Valera ◽  
J. Dorado ◽  
E. Terán ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Quality ◽  
Reproductive Traits ◽  
Study Groups ◽  
Inbreeding Coefficient ◽  
Computer Assisted ◽  
Effective Population ◽  
Mean Velocity ◽  
Sperm Analysis ◽  
Head Displacement

Reproductive performance is one of the key factors in livestock production. It is well known that reproductive traits are influenced by several genetic factors, such as the increase of individual inbreeding levels, which are associated with changes in sperm motility and shape in several species. In horses, the increase in inbreeding is a common problem because of the reduction in effective population size and the increase in selection intensity observed in several breeds. However, studies assessing the effect of high levels of inbreeding on the sperm quality of stallions are scarce. In the present study, we aimed to determine the effect of increased inbreeding levels and age on the sperm motility patterns of Purebred Spanish horses (PRE). We performed kinetic characterisation of 557 sperm samples of 82 PRE stallions aged between 3 and16 years, using computer-assisted sperm analysis (Androvision™, Minitube). We evaluated 5 parameters in 6 different fields per sample: curved line velocity (VCL, µm/s), velocity average path (VAP, µm/s), velocity straight line (VSL, µm/s), amplitude of lateral head displacement (ALH, µm), and beat-cross frequency (BCF, Hz). We determined the pedigree-based inbreeding coefficient (Fped) based on ∼300,000 PRE pedigree records to evaluate the inbreeding effect. Individuals were separated into 2 groups: highly inbred (n=339) and lowly inbred (n=218) according to an F value of 12.5%. Differences between groups were analysed using a generalized linear model. The analysis did not show significant differences (P&gt;0.05) in the variables analysed with respect to the age of stallions. However, VAP, VCL, and AHL were lower in highly inbred than in lowly inbred animals (P&lt;0.05), suggesting less velocity and amplitude of head displacement. In the case of BCF, no significant differences (P&gt;0.05) were observed between the two study groups. In conclusion, age did not affect sperm quality parameters in the age group of stallions analysed. In addition, we demonstrated that high inbreeding coefficient reduced the mean velocity and trajectory pattern of spermatozoa in PRE.

20 Assessment of Motion and Kinematic Characteristics of Semen from Four Cattle Breeds Using Computer-Aided Sperm Analysis

2018 ◽  
Vol 30 (1) ◽  
pp. 149
Author(s):  
M. L. Mphaphathi ◽  
M. M. Seshoka ◽  
T. R. Netshirovha ◽  
Z. C. Raphalalani ◽  
N. Bovula ◽  
...  
Keyword(s):  
Objective Evaluation ◽  
Semen Parameters ◽  
Sperm Cells ◽  
Sperm Analysis ◽  
Progressive Motility ◽  
Cattle Breeds ◽  
Bull Semen ◽  
Head Displacement ◽  
Computer Aided ◽  
Sperm Motion

Subjective semen evaluation using standard optical microscopy is the most common practice. Semen parameters routinely assessed are volume, concentration, progressive motility, and morphology. However, computer-aided sperm analysis (CASA) represents an objective evaluation, sperm assessment that are reproducible and reliable. Such semen parameters have not been evaluated in Afrikaner, Brahman, and Bonsmara bulls’ semen. The present study evaluated the sperm motion and kinematics characteristics of semen from stud Afrikaner, Brahman, Bonsmara, and Nguni bulls using CASA technology. The electro-ejaculator was used for semen collection from Afrikaner (n = 11), Brahman (n = 7), Bonsmara (n = 10) and Nguni (n = 16) bulls of known and proven fertility. Semen was collected following 4 days of resting period. The bulls ranged between 5 and 6 years of age. After collection, the semen samples were immediately transferred to a thermo-flask and maintained at 37°C for further evaluation in the mobile laboratory (Nedambale, 2014). The CASA-Sperm Class Analyzer® system (Microptic, Barcelona, Spain) was used to evaluate sperm motion, velocity, and kinematic parameters or characteristics of raw/fresh semen from 4 cattle breeds. Data were analysed using GenStat® statistical programme (VSN International, Hemel Hempstead, United Kingdom). Treatment means were compared using one-way ANOVA. The total sperm motility rate was similar for all breeds: Afrikaner (92.2 ± 4.2), Brahman (90.7 ± 9.0), Bonsmara (93.9 ± 4.0), and Nguni (96.0 ± 2.7). However, Brahman and Afrikaner bull semen had higher sperm cells moving in a progressive motility of 57.3 and 45.6%, respectively, compared with other breeds (P < 0.05). Nguni, Afrikaner, and Bonsmara had the highest sperm cells moving in a rapid movement of 73.7, 72.4, and 67.4% (P > 0.05), respectively. The bulls sperm trajectories had a variation, as they were recorded to be irregular and not linear (P < 0.05). The straight-line sperm velocity (µm s−1), wobbling %, and amplitude of lateral head displacement % was similar for the 4 breeds (P > 0.05). In conclusion, CASA technology was a useful technique for assessing differences in sperm motion and kinematic (motility and velocity characteristics) among different bull breeds.

scholarly journals Two Protocols of Cryopreservation of Goat Semen with the Use of Computer-Assisted Semen Analysis System

2007 ◽  
Vol 76 (4) ◽  
pp. 601-604 ◽  
Author(s):  
R. Kozdrowski ◽  
A. Dubiel ◽  
W. Bielas ◽  
M. Dzięcioł
Keyword(s):  
Citric Acid ◽  
Semen Analysis ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Semen Cryopreservation ◽  
Velocity Amplitude ◽  
Head Displacement ◽  
Analysis System ◽  
Thawing Procedure ◽  
Lateral Head

The objective of the study was a comparison of two protocols of goat semen cryopreservation with the use of computer-assisted semen analysis system. Twenty ejaculates obtained with electroejaculation method were assessed. Each ejaculate was divided in half and frozen according to two protocols. In protocol I semen was centrifuged in order to remove its plasma and diluted in Tris buffer extender containing glucose, citric acid and glycerol with 20% addition of egg yolk. Protocol II did not include removal of plasma and the extender contained 1.5% egg yolk. It was shown that the removal of semen plasma improved motility of goat spermatozoa following freezing/thawing with respect to the following motility indicators: motility, average path velocity, amplitude of lateral head displacement at p < 0.05, and straight velocity, straightness and linearity at p < 0.01. In conclusion, the removal of semen plasma through centrifugation improved motility properties of goat semen following the freezing/thawing procedure.

Sign in / Sign up

Export Citation Format

Share Document