ABSTRACT
The objective of this study was to investigate the effects of oocyte selection using brilliant cresyl blue (BCB) and culture density during individual in vitro maturation (IVM) on porcine oocyte maturity and subsequent embryo development using a chemically defined medium. Cumulus-oocyte complexes (COCs) were classified as BCB-positive or BCB-negative after exposure to a BCB solution for 90 min. The classified COCs were matured in a group (15 COCs per 100-microL droplet) or individually (1 COC per 1-, 2.5-, 5-, or 10-microL droplet). Meiotic competence, intraoocyte glutathione concentration, and developmental competence after intracytoplasmic sperm injection were monitored. The BCB selected oocytes competent for nuclear and cytoplasmic maturation. Furthermore, meiotic competence for oocytes matured individually in a 5-microL droplet was superior (P<0.05) to that of oocytes matured in a 1-microL droplet. Also, the culture density in a 5-microL droplet during IVM resulted in a higher (P<0.05) rate of cleaved embryos than that in a 1-microL droplet and produced a similar rate of blastocysts compared with that of a group culture system. Conversely, BCB selection did not improve cleavage and blastocyst formation. In conclusion, it was possible to predict porcine oocytes competent for maturation using oocyte selection with BCB. Moreover, a 5-microL droplet during the individual IVM culture was most suitable for oocyte maturation and subsequent embryo development, although every culture density used in this study supported development up to the blastocyst stage.
Subject(s)
Coloring Agents , Oocytes/cytology , Oocytes/growth & development , Oxazines , Swine , Animals , Blastocyst/physiology , Cell Separation/methods , Cell Separation/veterinary , Cells, Cultured , Culture Media , Embryo Culture Techniques/veterinary , Embryonic Development , Female , Glutathione/analysis , Male , Meiosis , Oocytes/chemistry , Ovarian Follicle/cytology , Sperm Injections, Intracytoplasmic/veterinaryABSTRACT
BACKGROUND: Previous studies have demonstrated appreciable tumor induction in mouse skin by daily irradiation with high-power long-wavelength ultraviolet A (UVA). OBJECT: The aim of the present study was to examine the enhancing effects of UVA on changes in mouse skin mediated by the tumor promoter 12-o-tetradecanoylphorbol-13-acetate (TPA) by measurement of ornithine decarboxylase (ODC) activity and morphometric analysis. In addition, we examined the inhibitory effects of curcumin, a component of turmeric, on these changes. METHOD: ODC activity in the epidermis of CD-1 mice was determined by the method of Russell and Snyder. Epidermal and dermal thickness, and the number of dermal infiltrating inflammatory cells were quantified using a computer-assisted image analyzer. RESULTS: A combination of topical TPA application and UVA irradiation produced a greater increment of ODC activity at 4 h than TPA alone (p < 0.05). Histopathologically, TPA plus UVA tended to increase the dermal infiltrating inflammatory cells in contrast to TPA alone. Pretreatment of mice with curcumin significantly abrogated the TPA-induced changes in ODC activity and the dermal infiltrating inflammatory cells as well as the TPA plus UVA-mediated enhancement of these changes. CONCLUSION: Our data indicate that UVA irradiation (18.72 J/cm2) significantly enhances ODC induction at an early stage (4-6 h) after topical application of TPA, and aggravates the dermatitis elicited by TPA. Pretreatment with curcumin significantly inhibits these enhancing effects.
