• Korean Journal of Animal Reproduction
• /
• v.22 no.4
• /
• pp.411-417
• /
• 1998

The objective of this study was to dertermine the effects of oviductal fluid and oviductal conditioned medium on polyspermy and in vitro development of porcine oocytes. The addition of oviductal fluid and oviductal conditioned medium in the prefertilization and fertilization medium significantly decreased polyspermy rates and the mean number of spermatozoa in penetrated eggs(P＜0.05). The acrosome reaction rate significantly increased when spematozoa were exposed for 1.5, 3, 4.5h in oviductal fluid and oviductal conditioned medium(P＜0.05). When oocytes cultured for 192h, the percentage of oocytes that developed to the morula and blastocyst stage was higher in culture medium with oviductal fluid and oviductal conditioned medium than without oviductal fluid and oviductal conditioned medium(P＜0.05). These results indicated that the oviductal secreations will effectively reduce both the polyspermy rates and the mean number of spermatwa in penetrated eggs. And the presence of culture with oviductal fluid and oviductal conditioned medium promotes in vitro development of porcine oocytes.

• Asian-Australasian Journal of Animal Sciences
• /
• v.22 no.7
• /
• pp.962-968
• /
• 2009

The aim of this study was to compare the effects of oviductal fluid, porcine zygote medium (PZM)-3, PZM-4 and PZM-5, and modified PZM-5 culture media, and determine the effects of zona pellucida (ZP) removal on the development of nuclear transfer (NT) embryos. There were no significant differences in the rates of fusion and cleavage among the five different oviductal fluid concentrations. However, the rates of blastocyst formation and the cell numbers per blastocyst were high in the embryos at the 14 and 28 $\mu{g}$/ml concentrations of oviductal fluid compared to the 0, 56 and 100 $\mu{g}$/ml concentrations. The rates of cleavage and blastocyst formation, and the cell numbers per blastocyst were higher in the PZM-3, PZM-5 and modified PZM-5 media than in the PZM-4 medium. However, there were no significant differences in the fusion rates of oocytes among the four culture media. The cell numbers per blastocyst in the embryos without ZP were significantly greater than those with ZP. However, there were no significant differences in the rates of fusion, cleavage and blastocyst formation between the embryos with and without ZP. In conclusion, we improved blastocyst development and the quality of NT embryos by replacing PVA with 3 mg/ml of BSA in PZM-5 medium and supplementing the PZM-5 medium with 14 $\mu{g}$/ml oviductal fluid. The NT embryos produced by the zona-free NT method had a high rate of blastocyst formation in the modified PZM-5 medium.

• Korean Journal of Animal Reproduction
• /
• v.20 no.1
• /
• pp.1-8
• /
• 1996

The objective of this study was to determine effects of oviductal fluid on the sperm penetration and subsequent in vitro development of porcine oocytes. The addition of oviductal fluid to the fertilization medium decreased sperm pen etration and the mean number of spermatozoa in penetrated eggs. The number of spermatozoa firmly bound to zona pellucida was also decreased in the presence of oviductal fluid. Chlortetracy cline (CTC) fluorescence patterns were used to determine incidence of capacitation and acrosome reaction. The proportion of capacitated a and acrosome free spermatozoa increased when spermatozoa were exposed for 1.5 and 3 h to oviductal fluid. These results suggest that the factor(s) in secretion from the oviduct reduces polyspermic fertilization and the number of spermatozoa that will penetrate porcine oocytes. The reduction of polyspermic penetration by oviductal secretions may be due to a reduced number of spermatozoa in the fertilization me-dium into an intact acrosome.

• Asian-Australasian Journal of Animal Sciences
• /
• v.11 no.1
• /
• pp.60-64
• /
• 1998

The different developmental stage embryos and oviductal and uterine fluids of rabbit were analyzed by gas chromatography. Myristic (C 14:0), palmitic (C 16:0), palmitoleic (C 16:1), stearic (C 18:0), oleic (C 18:1), linoleic (C 18:2), linolenic (C 18:3), arachidic (C 20:0), arachidonic (C 20:4), docosahexaenoic (C 22:6) and lignoceric (C 24:0) acids were the common fatty acid constituents with little exception. In most of the samples palmitic, oleic, linoleic and arachidonic acids were observed in high concentration. Moreover, linoleic, linolenic and arachidonic acids were the three poly-unsaturated fatty acids in both type sample except day-1 oviductal fluids. Similarly, in both day-1 and day-2 oviductal and uterine fluids myristic, palmitoleic, stearic, linolenic, arachidic and docosahexaenoic acids were in less composition or undetected.

• Asian-Australasian Journal of Animal Sciences
• /
• v.10 no.5
• /
• pp.523-527
• /
• 1997

The fatty acid compositions of oocytes, follicular, oviductal and uterine fluids of pig and cow were analyzed using gas chromatography. Myristic (C 14: 0), palmitic (C 16: 0), palmitoleic (C 16: 1), stearic (C 18 : 0), oleic (C 18: 1), linoleic (C 18: 2), linolenic (C 18: 3) and arachidonic (C 20: 4) acids were identified as the common fatty acid constituents with little exception. Oleic acid composition was the highest (21.90 to 36.24%) in both pig and cow followed by palmitic (18.61 to 31.90%) and stearic (10.34 to 20.39%) acid. The three polyunsaturated fatty acids like linoleic, linolenic and arachidonic acids were detected in both pig and cow reproductive fluid samples. Myristic acid was not detected in pig oviductal fluid. Similarly, in cow oocytes myristic, palmitoleic and linolenic acids were not detected. Moreover, palmitic, stearic, oleic and linoleic acid comprised about 80% (73.74 to 88.00%) of the total fatty acids in the different samples analyzed in both animals.

• Korean Journal of Animal Reproduction
• /
• v.22 no.2
• /
• pp.177-186
• /
• 1998

While tubal pregnancy is frequently observed in human, it has been reported to rarely occur in other mammals. To investigate the reason of the absence of tubal pregnancy in other mammals, the ability of bovine tubal(oviductal) fluid to su, pp.rt the outgrowth of mouse embryos waw examined by using an in vitro model system wherein the trophoblast cells of hatched mouse blastocysts attach to and outgrow on tissue culture plates coated with FBS. When mouse blastocysts grwon in vitro from 2-cell embryos were cultrued in the dishes coated with FBS, human follicular fluid(hFF) and bovine follicular fluid(bFF), respectively, underwent outgrowth by spreading onto the plastic dishes during 48 hr. In contrast, none of the embryos cultured in the dishes coated with BSA or bovine obiductal fluid(bOF) did outgrow but remained as late blastocysts. Since addition of bOF at 5mg/ml or higher conc. to the culture medium resulted in degeneration of all embryos during 48 hr culture, 10mM conc. of glutathione(GSH) was added to the bOF-containing medium to circumvent the toxicity of bOF. In addition, bOF was heated $65^{\circ}C$ for 30 min(hbOF) to get rid of its precipitating properties and then added to the culture medium. When blastocysts were cultured in the presence of both hbOF and GSH 45.4% of embryos attached to the culture dishes. However, none of these embryos underwent outgrowth. Fially embryos were cultured in the presence of both hbOF and GSH but in the dishes coated with FBS. When they were examined after 48 hr, all of the blastocysts exhibited well-developed outgrwoth. Based upon these results, it is concluded that bovine oviductal fluid is capable of su, pp.rting the attchment of mouse blastocysts onto the culture plaste whereas it cannot promote the outgrwoth of mouse blastocysts in vitro, probably due to the lack of outgrwoth factor.

• Development and Reproduction
• /
• v.5 no.1
• /
• pp.23-33
• /
• 2001

When mammalian oocytes ovulate into the oviduct, associating follicular fluid components are exposed to the oviductal environment, possibly resulting in the mutual interaction between fillicu1ar and oviductal fluids. In the Present study, we have demonstrated for the first time that components of fallicular fluid could be modified by the oviductal fluid. Gelatin zymographic analyses of human follicular fluid (hFF) obtained from IVF patients showed consistently the presence of 110 kDa gelatinase (GA110) in addition to many bands among which 62 kDa gelatinase was predominant. Addition of EDTA or phenanfhroline to the gelatinase substrate buffer during gel incubation abolished GA110 band whereas phenylmethylsulffnyl fluoride (PMSF) did not. In contrast, bovine oviductal fluid(bOF) exhibited only 62 kDa gelatinase. Surprisingly, when bOF was added to hFF in 1:1 ratio and then the mixture was incubated for 3 h at 37$^{\circ}$C, GA110 of hFF disappeared. Disappearance of GA110 by bOF was observed even within 30 min after mixing with hFF. Addition of aminophenylmercuric acetate (APMA) to hFF also abolished enzymatic activity of GA110 but increased the activityof 62 kDa gelatinase. However, APMA abolished many other gelatinases as well unlike bOF. Interestingly, treatment of hFF with EDTA for 3 h remarkably increased the enzymatic activity of GA110 but not that of other gelatinases. Addition of phenanthroline, PMSF or soybean trypsin inhibitor (SBTI) did not affect overall gelatinase activities. Again, addition of bOF to the hFF pretreated with any of the above proteinase inhibitors abolished the appearance of GA110. Human serum also showed GAI 10 of which activity was greatlyenhanced by EDTA treatment. Similar to hFF, serum GA110 also disappeared by the addition of bOF. Human granulosa cell homogenate did not reveal any appreciable gelatinase activity except 92 kDa gelatinase. Anti-human gelatinase A antibody reacted with 62 kDa gelatinase of hFF. Based upon these results, it is concluded that bOF could selectively degrade an isoform of gelatinase A present in hFF and human serum.

• Korean Journal of Animal Reproduction
• /
• v.18 no.4
• /
• pp.285-297
• /
• 1995

The mammalian oviduct is a place where ontogeny of an animal begins. Nowadays, however, it is possilbe to manipulate a part of physiological events occurring in the oviduct so that fertilization of gametes and early embryonic development of zygotes could proceed outside oviductal environment. Rabbit zygotes readily develop to blastocysts in a conventional culture condition. Most of the mouse fertilized eggs do so when cultured under a specific environment, e.g., in a medium containing ethylenediamine tetraacetic acid. Similarly, a significant number of zygotes from rat, sheep, pig or cattle can develop to blastocysts if they are cultured in the presence of particular component which appear to be somewhat species-specific. Instead of changing the components of medium, somatic cells including oviductal epithelial cells, have widely been used to improve mammalian embryonic development in vitro. Many investigators have reported that mammalian zygotes, whether fertilized in vivo or in vitro, could develop to blastocysts when they were cultured on a monolayer of various kinds of somatic cells or even in a somatic cell-conditioned medium. While little is known about the nature of embryotrophic factor(s) produced in vitro by somatic cells, the existence fo oviduct-specific protein(s) has consistently been demonstrated in many laboratories. Some of these proteins are reported to be associated with oviductal eggs. However, the physiological role of these proteins has still to be determined. Recently we observed that the perivitelline space of mouse oocytes was fluorescently stained with various fluorochrome-protein conjugates following ovulation into the oviducts or upon their expossure to oviductal extracts. Furthermore, it was also found that cattle or pig oviductal fluid gave similar results when examined using mouse ghost ZP. These observations lead to suggest that mammalian oviduct induces changes of biochemical properties of oocytes. Further studies are needed to clarify the nature of oviductal factor(s) and the physiological meaning of the reaction.

• 대한생식의학회:학술대회논문집
• /
• 1996.11a
• /
• pp.61.1-61.1
• /
• 1996

• 대한생식의학회:학술대회논문집
• /
• 1996.11a
• /
• pp.61.2-62
• /
• 1996