Browsing by Subject "Sperm"
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Item Assessment of boar sperm samples by computer-assisted sperm anaylsis and the mobility assay(2007-05) Tilley, Breanna; Vizcarra, Jorge A.; Alvarado, Christine Z.; Glayean, MichealThe sperm mobility assay (Accudenz) used in the present study measures sperm penetration into a biologically inert gradient solution. When a sample of sperm is overlaid in a cuvette containing Accudenz, sperm penetrate the solution and then measured with a spectrophotometer. The mobility assay has been successfully used to select chicken and turkey sperm donors. We validated this assay for semen from boars. Absorbance was measured after overlaying stored semen (24h) from each boar in prefilled cuvettes for 1, 5, 10, 15, 20, 40 min. There were no significant differences between sperm concentrations 1 x 108 and 5 x 107 viable sperm/mL. Absorbance was half-maximal at 13.1 min. In addition, there was medium repeatability for individual boars. There were positive correlations between mobility values and several computer-assisted sperm analysis (CASA) measurements, and low to high reproducibility for several CASA measurements. We concluded that further study is needed to find the proper Km and, the mobility and CASA measurements are correlated.Item Effect of Density Gradient Centrifugation on Quality and Recovery Rate of Equine Sperm(2010-07-14) Edmond, Ann J.Density gradient centrifugation of sperm is a common assisted-reproduction procedure in humans used to improve semen quality. The technique allows sperm separation based on their isopycnic points. Sperm with morphologic abnormalities are often more buoyant, leading to their retention above centrifuged density gradients, with structurally normal sperm passing through the gradient. Three experiments were conducted to evaluate the effects of tube size, sperm number following centrifugation, and density gradient volume (height) on stallion sperm quality and recovery rate in sperm pellets following centrifugation. In all three experiments, equine semen was initially centrifuged to increase sperm concentration. In Experiment 1, one-mL aliquots were layered over EquiPure? Bottom Layer (1-Layer) or over-tiered EquiPure? Top and Bottom Layers (2-Layer). For Experiment 2, one-mL aliquots were layered over three different heights of EquiPure? Bottom Layer in 15-mL or 50-mL conical-bottom tubes. For Experiment 3, four different aliquots containing a sperm load of 1-4x were layered over a constant volume of EquiPure? Bottom Layer in 15-mL or 50-mL conical bottom tubes. The tubes were then centrifuged. Resulting sperm pellets were evaluated for morphologic quality, DNA integrity, motility and recovery rate. Sperm-EquiPure? centrifugation yielded improvements in motility, morphology and DNA integrity parameters (P<0.05), as compared to controls. The 1-Layer method resulted in a higher recovery rate than the 2-Layer method (P<0.05). Sperm processed in the 15-mL tubes yielded higher velocity and higher recovery rates than sperm processed in the 50-mL tubes (P<0.05). Within tube type, gradient volume did not impact parameters of semen quality or recovery rate. An increase in sperm number for density gradient centrifugation resulted in a decreased recovery rate (P<0.05) when 15-mL tubes were used.Item Elucidating the signal cascades induced by progestins that mediate sperm hypermotility in Atlantic croaker (Micropogonias undulatus) and southern flounder (Paralichthys lethostigma)(2013-12) Tan, Wenxian, active 21st century; Thomas, P. (Peter)The overall goal of this research was to verify the involvement of membrane progestin receptor alpha (mPRα) in mediating progestin-stimulated sperm hypermotility in the Atlantic croaker and southern flounder. Sperm motility in Atlantic croaker and southern flounder were tested with both the endogenous progestin, 17,20β,21-trihydroxy-4-pregnen-3-one (20β-S) or the selective mPRα agonist, 10-ethenyl-19-norprogesterone (Org OD 02-0). In croaker, the Pi3k/Akt/Pde and ErbB2/Mapk intracellular signaling pathways were examined. The role of mPRα in mediating sperm hypermotility and fertility in southern flounder was also studied. The effects of seasonal hypoxia on sperm motility in croaker were investigated in a field study in the northern Gulf of Mexico in the fall of 2010. Finally, the effects of acidified activator solution (simulating ocean acidification) were studied in the laboratory. In vitro, Org OD 02-0 mimicked the stimulatory actions of 20β-S in inducing sperm hypermotility and intracellular signaling cascades in croaker and flounder sperm, indicating that mPRα is the mediator of progestin signaling in the sperm of these species. In croaker sperm, both the Pi3k/Akt/Pde and ErbB2/Mapk intracellular signaling pathways were shown to be important mediators of progestin-induced sperm hypermotility, suggesting novel functions of G [subscript olf] βγ-subunits in teleost sperm. In flounder sperm, mPRα was shown to be important in mediating sperm hypermotility as only high motility sperm with high expression of mPRα were responsive to progestin stimulation, resulting in higher fertilization success compared to low motility sperm. A single LHRHa injection resulted in increased sperm motility and fertility, associated with an increase in mPRα expression in the sperm plasma membrane. The results also suggest that the mPRα/Acy/cAMP pathway first described in croaker sperm is present in flounder sperm. Field studies of male Atlantic croaker exposed to chronic seasonal hypoxia showed that hypoxia exposure resulted in smaller gonads, lower spermatogenesis, reduced testicular mPRα expression, and in some sites, reduced sperm motility. Studies with croaker sperm using acidified activator solution to simulate ocean acidification indicated that croaker sperm were sensitive to environmental insult. Furthermore, the results suggested that the progestin signaling mechanism is more sensitive to changes in ocean pH levels than the mechanism that controls sperm motility.