BIOCHEMICAL STATUS OF BOAR SPERMATOZOA and SEMINAL PLASMA BEFORE and AFTER A 10-DAY DEPLETION TEST (DT)

1995 ◽  
Vol 31 (1) ◽  
pp. 245-246 ◽  
Author(s):  
J. Strzezek ◽  
W. Demianowicz ◽  
W. Kordan ◽  
J. Torska ◽  
P. Wysocki ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Biochemical Status ◽  
Before And After ◽  
Boar Spermatozoa

Colloid centrifugation removes seminal plasma and cholesterol from boar spermatozoa

2011 ◽  
Vol 23 (7) ◽  
pp. 858 ◽  
Author(s):  
R. Kruse ◽  
P. C. Dutta ◽  
J. M. Morrell
Keyword(s):  
Plasma Membrane ◽  
Seminal Plasma ◽  
Plasma Proteins ◽  
Single Layer ◽  
Sperm Cryopreservation ◽  
Before And After ◽  
Sperm Plasma Membrane ◽  
Seminal Plasma Proteins ◽  
Boar Spermatozoa ◽  
Layer Chromatography

The objective of the present study was to investigate the effect of Single-Layer Centrifugation (SLC) on boar spermatozoa, namely the effect of removal of seminal plasma proteins and cholesterol from the surface of spermatozoa. The presence of porcine seminal plasma proteins I and II (PSP-I/PSP-II) before and after SLC was studied using immunofluorescence, whereas the removal of cholesterol was shown qualitatively by thin-layer chromatography (TLC). Finally, the integrity of the sperm plasma membrane was observed by electron microscopy. It was shown that the seminal plasma proteins PSP-I and -II were removed from spermatozoa during SLC but could be restored by adding seminal plasma to the SLC-selected sperm samples. Some cholesterol was also lost from the spermatozoa during SLC but the plasma membrane itself appeared to be morphologically intact. Further studies are underway to examine the relevance of these findings to boar sperm cryopreservation and sperm fertility.

Changes in epididymal protein anti-agglutinin on ejaculated boar spermatozoa during capacitation in vitro

1999 ◽  
Vol 11 (5) ◽  
pp. 193 ◽  
Author(s):  
Hiroshi Harayama ◽  
Masashi Miyake ◽  
Susan F. Magargee ◽  
Elaine Kunze ◽  
Seishiro Kato ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Western Blotting ◽  
Seminal Plasma ◽  
Indirect Immunofluorescence ◽  
Subcutaneous Injection ◽  
Early Stage ◽  
Before And After ◽  
Purified Antigen ◽  
Boar Spermatozoa

This study is a detailed investigation of changes in epididymal protein anti-agglutinin on ejaculated boar spermatozoa during an incubation designed to promote capacitation in vitro. Ejaculated spermatozoa were collected from six mature boars, washed, and incubated to promote capacitation. Sperm samples were subjected to Western blotting-densitometric analyses, flow cytometry after immunostaining and immunocytochemical observation by indirect immunofluorescence. An antiserum to anti-agglutinin was raised in a rabbit by subcutaneous injection of a purified antigen, as described previously (Harayama et al. 1999). Western blotting-densitometric analyses revealed an approximate halving of the amount of sperm-bound anti-agglutinin during the first 45-min incubation, followed by a gradual decrease thereafter. Comparison between immunostained sperm samples by flow cytometry before and after incubation confirmed this decrease in sperm-bound anti-agglutinin during the incubation. Microscopic characterization established that this decrease occurred mainly on the acrosome. Supplementation with seminal plasma (5% or 10%, v/v) attenuated the decrease. These findings are consistent with the conclusion that a large portion of the anti-agglutinin bound to sperm acrosomes is released at an early stage of the capacitation process in vitro.

scholarly journals Effects of Seminal Plasma Components on Motility and Acrosomal Integrity of Meishan Boar Spermatozoa after Cooling Treatments

1998 ◽  
Vol 69 (8) ◽  
pp. 720-727
Author(s):  
Hiroshi HARAYAMA ◽  
Akira IMANO ◽  
Masashi MIYAKE ◽  
Seishiro KATO
Keyword(s):  
Seminal Plasma ◽  
Boar Spermatozoa ◽  
Acrosomal Integrity

scholarly journals Changes in Bull Semen Metabolome in Relation to Cryopreservation and Fertility

Animals ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 1065
Author(s):  
Valentina Longobardi ◽  
Michal A. Kosior ◽  
Nunzia Pagano ◽  
Gerardo Fatone ◽  
Alessia Staropoli ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Glycine Betaine ◽  
Low Fertility ◽  
High Fertility ◽  
Over Expression ◽  
Bull Semen ◽  
Before And After ◽  
Associated Proteins ◽  
Cellular Compartments

Semen cryopreservation determines several sperm damages, including the loss of fertility-associated proteins. The purpose of the study was to compare the metabolite contents in bovine sperm and seminal plasma before and after cryopreservation, and between high- and low-fertility bulls in vitro. Forty-eight ejaculates, collected from eight bulls (six per bull), were analyzed by liquid chromatography–mass spectrometry. Cryopreservation resulted in an over-expression of lysophosphatidylcholine (0:0/18:2(9Z,12Z)) in seminal plasma. In addition, higher levels of glycine betaine and pyro-l-glutaminyl-l-glutamine were observed in cryopreserved compared to fresh spermatozoa. The fresh seminal plasma of high-fertility bulls showed an over-expression of l-acetylcarnitine, glycerol tripropanoate, 2,3-diacetoxypropyl stearate and glycerophosphocholine, and an under-expression of lysophosphatidylcholine and butyrylcarnitine, compared to low-fertility bulls. Higher levels of glycerophosphocholine and lysophosphatidylcholine (16:0/0:0) were recorded in fresh spermatozoa from high-fertility bulls. In high-fertility bulls, a greater content of glycerophosphocholine and lower levels of butyrylcarnitine, glycine betaine and l-carnitine were found in cryopreserved seminal plasma, and lower levels of glycine betaine were detected in cryopreserved spermatozoa. In conclusion, cryopreservation affects bovine semen metabolome at both plasmatic and cellular compartments, and metabolic profile differs between high- and low-fertility bulls.

scholarly journals Induced hyperactivity in boar spermatozoa and its evaluation by computer-assisted sperm analysis

Reproduction ◽  
2004 ◽  
Vol 128 (2) ◽  
pp. 171-179 ◽  
Author(s):  
Harald Schmidt ◽  
Günter Kamp
Keyword(s):  
Seminal Plasma ◽  
Roc Curve ◽  
Curve Analysis ◽  
Computer Assisted ◽  
Ionophore A23187 ◽  
Threshold Values ◽  
Roc Curve Analysis ◽  
Sperm Analysis ◽  
Computer Assisted Sperm Analysis ◽  
Boar Spermatozoa

Hyperactivity, a form of sperm motility characterized by vigorous flagellar movements, has been proposed as essential for fertilization in mammals. The objective of the present study was to establish a method for inducing hyperactivityin vitroin boar spermatozoa and to define threshold values to differentiate between hyperactive and non-hyperactive spermatozoa by computer-assisted sperm analysis (CASA) as a prerequisite for analyzing the energy metabolism during hyperactivity. In TALP-HEPES medium, non-frozen boar spermatozoa were stimulated to hyperactivity by 50 μmol l−1Ca2+within 15 min at 37 °C if 5 μmol l−1of the Ca2+ionophore A23187 was present. If 25% seminal plasma was present, boar spermatozoa required higher Ca2+concentrations (about 700 μmol l−1) for hyperactivity. Under both conditions, immobilization and head-to-head agglutination were low so that hyperactive spermatozoa could be analyzed for at least 40 min. The transition from normal to hyperactive movement was characterized by an increase in flagellar beat angle from 49° ± 12° to 200° ± 36° (n= 32) and a decrease in flagellar curvature ratio from 0.89 ± 0.04 to 0.47 ± 0.11 (n= 32). For quantification of hyperactive boar sperm, kinematic parameters of hyperactive and non-hyperactive spermatozoa were measured by CASA and statistically evaluated (receiver operating characteristic (ROC) curve analysis). The threshold values of the following four parameters were well suited for differentiating between hyperactive and non-hyperactive boar spermatozoa (ROC curve analysis: >50% specificity at 100% sensitivity). Hyperactive boar spermatozoa showed mean lateral head displacement >3.5 μm, curvilinear velocity >97 μm s−1, linearity <32% and wobble <71%. According to this multiparametric definition, induction of hyperactivity increased significantly (P< 0.0001) the fraction of hyperactive spermatozoa in semen samples from 5.1 ± 4.3% (n= 13) to 48.3 ± 6.6% (n= 7) in the absence and to 44.2 ± 7.6% (n= 10) in the presence of 25% seminal plasma, while the overall percentage of motile spermatozoa did not change significantly.

92 EVALUATION OF BOAR SPERM FUNCTIONALITY AFTER A CUSHIONED CENTRIFUGATION TECHNIQUE

2006 ◽  
Vol 18 (2) ◽  
pp. 154
Author(s):  
J. Gadea ◽  
S. Martínez-Miró ◽  
G. Decuadro-Hansen ◽  
C. Matás
Keyword(s):  
Seminal Plasma ◽  
Acrosome Reaction ◽  
Sperm Viability ◽  
Lipid Packing ◽  
Lipid Disorder ◽  
Merocyanine 540 ◽  
Centrifuge Tube ◽  
Before And After ◽  
Centrifugation Technique

Separation of sperm from seminal plasma is required in most semen freezing procedures. Semen is typically subjected to centrifugation to concentrate sperm into a pellet and allow removal of the seminal plasma prior to dilution in freezing extender. Centrifugation is a relatively effective method to recover sperm, however, the process also causes considerable sperm damage. The use of a dense, inert, and isotonic solution as a cushion in the bottom of the centrifuge tube allows a greater centrifugation speed to be applied and results in greater sperm recovery. The aim of the present work was to evaluate the effects of this cushioned centrifugation technique on in vitro sperm viability and functionality. Sperm-rich fractions from 16 fertile boars were diluted and cooled to 15�C; then subsamples were centrifuged by one of two different techniques. A standard method (SM), 800 g for 10 min in 50-mL tubes (Westendorf et al. 1975 Dtsch. Tier�rztl. Wschr. 82, 261-267) and a cushioned method (CM), 1000 g for 20 min using 45 mL of diluted semen on 5 mL of an isotonic iodixanol solution (60% w/v gradient) were performed. Sperm samples were stained with merocyanine 540 (M540) and Yo-Pro 1 (Harrison et al. 1996 Mol. Rep. Dev. 45, 378-391) to detect changes in lipid packing disorder of the plasma membrane. Another set of sperm samples was incubated in the presence of (0.7 �M) 22,72-dichlorodihydrofluorescein diacetate (Gadea et al. 2005 J. Androl. 26, 396-404) to estimate production of reactive oxygen species (ROS). A final set of sperm samples was stained with peanut aggultinin-fluorscein isothiocyanate (PNA-FITC) and propidium iodide to evaluate the acrosome reaction. All of these parameters were evaluated by flow cytometry before and after centrifugation. ANOVA analysis revealed that centrifugation altered lipid packing disorder and viability. Raw semen (RS) had a larger number of viable low lipid disorder sperm than centrifuged semen (RS = 86.9a vs. SM = 81.64b vs. CM = 80.6b, P < 0.01) and a decreased number of dead sperm cells (RS = 9.5a vs. SM = 15.0b vs. CM = 16.3b, P < 0.01). However, the cushioned and standard centrifugation methods yielded similar results for all the parameters measured. No significant differences were found for generation of ROS or in the number of sperm exhibiting the acrosome reaction. In conclusion, compared to the standard centrifugation method, this simple cushioned modification is a more efficient means of processing boar semen for freezing because significantly less sample losses are detected; also, it provides similar levels of sperm viability and functionality, and consequently a higher number of doses per ejaculation can be produced.

scholarly journals Quantitative Analysis of the Seminal Plasma Proteome in Secondary Hypogonadism

2019 ◽  
Vol 8 (12) ◽  
pp. 2128 ◽  
Author(s):  
Giuseppe Grande ◽  
Federica Vincenzoni ◽  
Francesca Mancini ◽  
Ferran Barrachina ◽  
Antonella Giampietro ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Seminal Plasma ◽  
Control Group ◽  
Testosterone Replacement Therapy ◽  
Male Hypogonadism ◽  
Grey Zone ◽  
Label Free ◽  
Clinical Markers ◽  
Before And After ◽  
Secondary Hypogonadism

In the grey zone of testosterone levels between 8 and 12 nmol/L, the usefulness of therapy is controversial; as such, markers of tissue action of androgens may be helpful in adjusting clinical decisions. To better understand the effect of the hypothalamic-pituitary-testicular axis on male accessory secretion, we performed a proteomic quantitative analysis of seminal plasma in patients with secondary hypogonadism, before and after testosterone replacement therapy (TRT). Ten male patients with postsurgical hypogonadotrophic hypogonadism were enrolled in this study, and five of these patients were evaluated after testosterone treatment. Ten men with proven fertility were selected as a control group. An aliquot of seminal plasma from each individual was subjected to an in-solution digestion protocol and analyzed using an Ultimate 3000 RSLC-nano HPLC apparatus coupled to a LTQ Orbitrap Elite mass spectrometer. The label-free quantitative analysis was performed via Precursor Ions Area Detector Node. Eleven proteins were identified as decreased in hypogonadic patients versus controls, which are primarily included in hydrolase activity and protein binding activity. The comparison of the proteome before and after TRT comes about within the discovery of six increased proteins. This is the primary application of quantitative proteomics pointed to uncover a cluster of proteins reflecting an impairment not only of spermatogenesis but of the epididymal and prostate epithelial cell secretory function in male hypogonadism. The identified proteins might represent putative clinical markers valuable within the follow-up of patients with distinctive grades of male hypogonadism.

scholarly journals From One Ejaculate to Another: Transference of Sperm Traits via Seminal Plasma Supplementation in the Ram

Biology ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 33
Author(s):  
Christine Green ◽  
Jessica P. Rickard ◽  
Simon P. de Graaf ◽  
Angela J. Crean
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Perceived Risk ◽  
Positive Relationship ◽  
Assisted Reproductive Technologies ◽  
Sperm Quality ◽  
Reproductive Technologies ◽  
Before And After ◽  
The Difference ◽  
Sperm Traits

Males can adjust sperm motility instantaneously in response to the perceived risk of sperm competition. The speed of this response suggests that sperm motility is regulated by changes in seminal plasma rather than changes in the sperm cells themselves. Hence, here we test whether inter-ejaculate variation in seminal plasma can be used to alter sperm quality prior to use in assisted reproductive technologies. We supplemented fresh ejaculates of Merino rams with seminal plasma collected from previous ‘donor’ ejaculates to test whether changes in sperm kinetics were related to the relative quality of donor to focal ejaculates. We found a positive relationship between the change in sperm traits before and after supplementation, and the difference in sperm traits between the donor and focal ejaculate. Hence, sperm motility can be either increased or decreased through the addition of seminal plasma from a superior or inferior ejaculate, respectively. This positive relationship held true even when seminal plasma was added from a previous ejaculate of the same ram, although the slope of the relationship depended on the identity of both the donor and receiver ram. These findings indicate that seminal plasma plays a key role in the control and regulation of sperm kinetics, and that sperm kinetic traits can be transferred from one ejaculate to another through seminal plasma supplementation.

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