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Acrosome Integrity (acrosome + integrity)
Selected AbstractsEffect of Docosahexaenoic Acid on Quality of Cryopreserved Boar Semen in Different BreedsREPRODUCTION IN DOMESTIC ANIMALS, Issue 3 2010K Kaeoket Contents During the cryopreservation process, the level of polyunsaturated fatty acids, especially docosahexaenoic acid (DHA), in the sperm plasma membrane decreases significantly because of lipid peroxidation, which may contribute to sperm loss quality (i.e. fertility) of frozen,thawed semen. The aim of this study was to investigate the effect of supplementation of DHA (fish oil) in freezing extender II on frozen,thawed semen quality. Semen from 20 boars of proven motility and morphology, were used in this study. Boar semen was split into four groups, in which the lactose,egg yolk (LEY) extender used to resuspend the centrifuged sperm pellet was supplemented with various levels of fish oil to reach DHA level of 1X (group I, control, no added fish oil), 6X (group II), 12X (group III) and 18X (group IV). Semen solutions were frozen by using a controlled rate freezer. After cryopreservation, frozen semen was thawed and evaluated for progressive motility, viability by using SYBR-14/Ethidiumhomodimer-1 (EthD-1) staining and acrosome integrity by using FITC-PNA/EthD-1 staining. There was a significantly higher (p < 0.001) percentage of progressive motility, viability and acrosome integrity in DHA (fish oil) supplemented groups than control group. Generally, there seemed to be a dose-dependent effect of DHA, with the highest percentage of progressive motility, viability and acrosome integrity in group-III. In conclusion, supplementation of the LEY extender with DHA by adding fish oil was effective for freezing boar semen as it resulted in higher post-thaw plasma membrane integrity and progressive motility. [source] Studies on the Effect of Supplementing Boar Semen Cryopreservation Media with Different Avian Egg Yolk Types on in Vitro Post-thaw Sperm QualityREPRODUCTION IN DOMESTIC ANIMALS, Issue 1 2006R Bathgate Contents Fertility after insemination of cryopreserved boar semen is currently below that of fresh semen. In an attempt to improve the post-thaw motility and acrosome integrity of boar sperm, semen was frozen using an adapted Westendorf method in which the chicken egg yolk was replaced by either duck or quail egg yolk. The different composition of the yolk types, particularly the amount of cholesterol, fatty acids and phospholipids, were thought to potentially afford a greater level of protection to sperm against damage during freezing and thawing. Sperm frozen in medium containing chicken egg yolk displayed higher motility immediately after thawing, but there was no difference in the motility of sperm frozen with different types of egg yolk 3 or 6 h after thawing and maintenance at 37°C. Sperm frozen in media containing chicken or duck egg yolk had a higher proportion of intact acrosomes immediately after thawing than sperm frozen in medium containing quail egg yolk, but 6 h after thawing and maintenance at 37°C the sperm that had been frozen in medium containing chicken egg yolk had a higher proportion of intact acrosomes than the sperm frozen in media containing duck or quail egg yolk. Analysis of the composition of the different yolk types showed that the basic components of the yolks were similar, but the ratios of fatty acids and phospholipid classes differed. Duck egg yolk had more monounsaturated fatty acids (MUFA) than chicken egg yolk, which had more MUFA than quail egg yolk. Duck egg yolk contained more phosphotidylinositol (PI) than chicken or quail egg yolks and quail egg yolk contained more phosphotidylserine than either chicken or duck egg yolks. The differences in post-thaw motility and acrosome integrity of boar sperm when frozen in media containing the different types of egg yolk may be due to the variation in composition. [source] Effect of amino acids on cryopreservation of cynomolgus monkey (macaca fascicularis) spermAMERICAN JOURNAL OF PRIMATOLOGY, Issue 4 2003Yahui Li Abstract The effects of three amino acids (proline, glutamine, and glycine) added to the freezing medium Tes-Tris-egg yolk (TTE) for cryopreservation of cynomolgus monkey (Macaca fascicularis) spermatozoa were studied. This is the first report on the effects of amino acids on nonhuman primate sperm cryopreservation. The addition of 5mM proline, 10mM glutamine, and 10 or 20mM glycine each significantly improved post-thaw sperm motility and membrane and acrosome integrity compared with the control (TTE alone). However, a significant decrease in motility and membrane/acrosome integrity was observed when amino acid concentrations increased to 60mM for proline and glutamine, and 80mM for glycine. The results suggest that adding a limited amount of amino acids to the freezing media is beneficial for freezing cynomolgus monkey sperm. Am. J. Primatol. 59:159,165, 2003. © 2003 Wiley-Liss, Inc. [source] Protein tyrosine phosphorylation under capacitating conditions in porcine fresh spermatozoa and sperm cryopreserved with and without alpha tocopherolANDROLOGIA, Issue 3 2009M. M. Satorre Summary The aim of this study was to evaluate the capacitation behaviour of fresh and ,-tocopherol frozen spermatozoa. Spermatozoa frozen with or without ,-tocopherol and fresh semen were incubated under capacitating conditions. Aliquots were collected at 0, 15, 30, 45, 60, 90, 120 and 180 min of incubation time. Parameters of semen quality were evaluated by optical microscopy and capacitation was determined by the epifluorescence chlortetracycline technique. Protein tyrosine phosphorylation was examined by Western immunoblotting. Motility, viability and intact spermatozoa were higher (P < 0.05) in fresh semen compared with frozen samples. These parameters significantly decreased, in every treatment, throughout the incubation time. Fresh semen showed a progressive increase in capacitated spermatozoa, reaching 25 ± 3% at 180 min. Cryopreserved semen had a fast increase at the beginning of incubation time (28 ± 5% at 45 min and 28 ± 3% at 30 min for samples with or without ,-tocopherol, respectively). The amount of an MW 32 kDa tyrosine-phosphorilated protein, associated with capacitation, increased throughout incubation for fresh semen and spermatozoa cryopreserved with ,-tocopherol. The supplementation with ,-tocopherol preserved sperm plasma membrane, reflected not only in the acrosome integrity but also in a greater efficiency of energy production. [source] Association between freezing agent and acrosome damage of human spermatozoa from subnormal and normal semenANDROLOGIA, Issue 6 2001M. E. Hammadeh Summary. This experimental study compares the effects of human sperm preservation medium (HSPM) with TEST,yolk buffer (TYB) as cryoprotectants of human spermatozoa with respect to the integrity of the acrosome after the freeze,thawing procedure. Fifty-six semen samples were included in this study; 18 were subnormal (G1) and 38 were normal (G2) based on World Health Organization criteria, except for morphology, which was evaluated according to strict criteria. Each semen sample was divided into two parts: the first part was prepared for cryopreservation by the addition of HSPM (1:1) and the second by addition of TYB (1:1). Freezing was performed in liquid nitrogen vapour. Smears were made before freezing and after the thawing process for evaluation of acrosome integrity using fluorescent-lectin labelling. The mean percentage of spermatozoa with intact acrosomes in the subnormal group was 77.0 ± 7.2% before freezing and decreased significantly (P < 0.001) after thawing: to 63.7 ± 8.2% with the use of HSPM and 66.8 ± 8.7% with the use of TYB. The corresponding values in the normal semen samples were 83.4 ± 9.2%, 76.0 ± 8.8% and 77.9 ± 9.2%, respectively. It is obvious that the decrease in the mean percentage of spermatozoa with intact acrosome was significantly higher when using HSPM in comparison with TYB, not only for G1 (,14.9 ± 1.9% versus ,11.8 ± 1.4%) but also for G2 samples (,13.8 ± 1.5% versus ,11.9 ± 1.3%). In conclusion, TYB should be recommended for freeze,thawing of human spermatozoa as the first-choice cryoprotectant, for normal as well as subnormal semen samples, in order to protect the sperm acrosome from the deleterious effects of the freeze,thawing procedure. [source] Do physical forces contribute to cryodamage?BIOTECHNOLOGY & BIOENGINEERING, Issue 4 2009Joseph Saragusty Abstract To achieve the ultimate goal of both cryosurgery and cryopreservation, a thorough understanding of the processes responsible for cell and tissue damage is desired. The general belief is that cells are damaged primarily due to osmotic effects at slow cooling rates and intracellular ice formation at high cooling rates, together termed the "two factor theory." The present study deals with a third, largely ignored component,mechanical damage. Using pooled bull sperm cells as a model and directional freezing in large volumes, samples were frozen in the presence or absence of glass balls of three different diameters: 70,110, 250,500, and 1,000,1,250,µm, as a means of altering the surface area with which the cells come in contact. Post-thaw evaluation included motility at 0,h and after 3,h at 37°C, viability, acrosome integrity, and hypoosmotic swelling test. Interactions among glass balls, sperm cells, and ice crystals were observed by directional freezing cryomicroscopy. Intra-container pressure in relation to volume was also evaluated. The series of studies presented here indicate that the higher the surface area with which the cells come in contact, the greater the damage, possibly because the cells are squeezed between the ice crystals and the surface. We further demonstrate that with a decrease in volume, and thus increase in surface area-to-volume ratio, the intra-container pressure during freezing increases. It is suggested that large volume freezing, given that heat dissipation is solved, will inflict less cryodamage to the cells than the current practice of small volume freezing. Biotechnol. Bioeng. 2009; 104: 719,728 © 2009 Wiley Periodicals, Inc. [source] | |||||||||||||