• Asian-Australasian Journal of Animal Sciences
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• v.14 no.10
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• pp.1360-1366
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• 2001

The aims of this study were first to evaluate the effects of different levels (20, 40 and 100%) and sources (follicular size: large, >7 mm; medium, >5-7 mm; small, 3-5 mm) of porcine follicular fluid (pFF) on the in vitro maturation (IVM) of porcine oocytes, and the effects of fertilization treatments and different culture conditions on development of fertilized oocytes were also investigated. No differences in the maturation (63.6-76.6%) and cleavage (24.8-34.3%) rates were observed among the 20,40 and 100% pFF groups (p>0.05). The cleavage rates of oocytes cultured and fertilized in 40% and 100% pFF maturation media were significantly higher than those fertilized in m199-NBCS (51.0-61.2% vs. 12.8-31.8%. p<0.05), regardless of sources of the pFF. When oocytes were fertilized in m199-NBCS followed by culture in rabbit oviducts for 4 days, the cleavage rate in 40% pFF group was better than that in 100% pFF group (46.9% vs. 32.5%, p<0.05). Two oocytes recovered from the oviducts in the 40% pFF group developed to blastocysts after IVC. However, none developed to blastocysts when fertilized in the IVM medium after being transferred to rabbit oviducts. In conclusion, addition of pFF accompanied with gonadotropins (FSH, LH) in IVM medium enhanced maturation and cleavage rates of porcine oocytes. Direct addition of sperm suspension to IVM medium may be an alternative to simplify the fertilization procedures and to reduce the mechanical lesion during manipulation. Furthermore, rabbit oviducts provide a better environment for the in vitro fertilized oocyte developing to the morula and blastocyst stages.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.4
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• pp.347-351
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• 2021

Objective: We investigated the impact of vitamin D₃ (VD3) supplementation during mouse preantral follicle culture in vitro and the mRNA expression of 25-hydroxylase (CYP2R1), 1-alpha-hydroxylase (CYP27B1), and vitamin D receptor (VDR) in mouse ovarian follicles at different stages. Methods: Preantral follicles were retrieved from 39 BDF1 mice (7-8 weeks old) and then cultured in vitro for 12 days under VD3 supplementation (0, 25, and 50 pg/mL). Follicular development and the final oocyte acquisition were assessed. Preantral follicles were retrieved from 15 other BDF1 mice (7-8 weeks old) and cultured without VD3 supplementation. Three stages of mouse ovarian follicles were obtained (preantral, antral, and ruptured follicles). Total RNA was extracted from the pooled cells (from 20 follicles at each stage), and then reverse transcriptase-polymerase chain reaction was performed to identify mRNA for CYP2R1, CYP27B1, and VDR. Results: The survival of preantral follicles, rates of antrum formation and ruptured follicles (per initiated follicle) and the number of total or mature oocytes were all comparable among the three groups. Both CYP2R1 and CYP27B1 were expressed in antral and ruptured follicles, but not in preantral follicles. VDR was expressed in all three follicular stages. Conclusion: VD3 supplementation in vitro (25 or 50 pg/mL) did not enhance mouse follicular development or final oocyte acquisition. Follicular stage-specific expression of CYP2R1, CYP27B1, and VDR was observed.

• Korean Journal of Ichthyology
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• v.19 no.1
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• pp.16-23
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• 2007

Oocyte surface in two Korean minnows, Rhynchocypris oxycephalus and R. kumgangensis was examined by light and electron microscope. In two species, the development of the oocyte was similar, but the follicular layer surrounding full-grown oocyte showed an evident difference. In R. oxycephalus, the follicular layer at the yolk vesicle stage became bilaminar with the retention of its outer squamous cell layer and the acquisition of an inner cuboidal or round cell layer just over the zona radiata. As the oocyte grows, the cuboidal cells of the inner follicular layer began to be replaced by columnar cells. At the yolk granule stage, the columnar cells secreted mucin to their cytoplasm (adhesive materials) and then surround the entire oocyte, as bundles of fence-shaped structures. Whereas, although the follicular layer of R. kumgangensis had an outer squamous layer and an inner cuboidal or round cell layer at the yolk vesicles as in R. oxycephalus, no inner cells were more changed with the retention of its cuboidal or round cells. Finally, in R. kumgangensis, the adhesive materials did not occur. In Korean two minnows, the structural difference in the oocyte surface seems to be related to their habitats and spawning characteristics as well as taxonomic characters.

• Korean Journal of Animal Reproduction
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• v.13 no.2
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• pp.79-84
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• 1989

This study was carried out to investigate the effect of LH-RH and Gn-RH treatment in Holstein cows with ovarian quiescence and follicular cystic ovaries. The cows with ovarian quiescence and follicular cystic ovaries injected intramuscularly with 100$\mu\textrm{g}$, 200$\mu\textrm{g}$ and 400$\mu\textrm{g}$ of LH-RH and 200$\mu\textrm{g}$ and 400$\mu\textrm{g}$ of Gn-RH respectively. The cows was diagnosed by repeated rectal palpation. The plasma progesterone and estradiol-17$\beta$ concentrations were assayed by radioimmunoassay methods. The resutls of this study were summarized as follows : 1. Ovulations were induced after treatment of LH-RH and Gn-RH. The concentrations of progesterone reached small peak level at luteal phase and estradiol-17$\beta$ reached obvious peak level with the development and maturation of the follicle during the periods of degeneration of the corpus luteum, and normal ovarian cycle activity started subsequently. 2. The cows with ovarian quiescence and follicular cystic ovaries were induced ovulation at 38.9$\pm$5.3 hrs. after treatment of LH-RH in 66.7% cows and at 52.7$\pm$7.9 hrs after treatment of Gn-RH in 60.0% cows respectively. 3. The good ovarian responses were indicated in treatment with 200$\mu\textrm{g}$ to 400$\mu\textrm{g}$ of LH-RH than those treated with 100$\mu\textrm{g}$ in cows with ovarian quiescence, and did not show difference of ovarian responses between treatments with 200$\mu\textrm{g}$ to 400$\mu\textrm{g}$ of Gn-RH in cows with follicular cystic ovaries.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.12
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• pp.1647-1651
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• 2004

The follicles (1.8 to 7.8 mm in diameter) were recovered from the ovaries in marketed pigs and the number of granulosa cells, the diameter of oocytes obtained from different development stages of the follicles and follicular fluid levels were determined. Correlations between size measurements and cell counts as well as the diameter of antral follicles and oocytes were also investigated. The results indicated that, while expanding in size, follicle numbers decreased with a greater atretic proportion. Granulosa cells increased in numbers continuously and remained unchanged beyond the size of 200 ${mm}^3$ in non-atretic follicles, whereas a sudden drop of granulosa counts was observed in atretic follicles. Follicular fluid, on the other hand, linearly increased its volume with follicle size and differed little between those of non-atretic and atretic follicles. Diameters of oocytes in non-atretic follicles increased to its maximum when follicles expanded to 150 ${mm}^3$ and maintained its size during later follicular expansion. It is concluded that, for in vitro culture, the optimal size of porcine follicle should be between 150 to 180 ${mm}^3$if they are collected from pre-pubertal gilts of marketing size slaughtered in an abattoir.

• Journal of Veterinary Clinics
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• v.24 no.4
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• pp.563-567
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• 2007

This study evaluated the effect of ANTORIN R-10(pFSH), a commercially available follicle stimulating hormone on ovarian morphology, on follicular development and serum estradiol levels in rats. Immature female Wistar S/T rats(27 day old; 80-100 g B.wt) maintained under controlled environmental conditions($22{\pm}2^{\circ}C$; 50% humidity; 12 h light/12 h dark cycle) with free access to standard laboratory feed and tap water were utilized. Animals were allowed to acclimatize to the new environment for at least 2 weeks before being included in the experiment. Rats were randomly allotted to 5 groups(Control, SL 0.1AU, SH 0.2AU, TL 0.1AU and TH 0.2AU). ANTORIN R-10 was subcutaneously injected twice daily for 3 days. Twenty hours after hormone treatment, blood was collected to estimate the serum estradiol $17-\beta$ concentration. Immediately, all rats were sacrificed and the ovarian morphology, ovary weight and number of follicles were recorded. Ovaries were fixed for histomorphological examination. Higher standard and treatment groups were significantly increased on ovary weight and the number of follicles more than 1mm compared with lower standard and treatment. However, no difference revealed between standard and treatment groups. ANTORIN R-10 was similar effects of follicles development and maturation compared with House standard FSH.

• Asian-Australasian Journal of Animal Sciences
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• v.8 no.2
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• pp.123-127
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• 1995

In vitro embryo production (IVP) is affected by various factors during in vitro maturation, fertilization, and development. In this experiment, the effect of ovary type, quality of follicular oocyte, medium used for fertilization, presence of hormone in medium, sperm concentration on in vitro maturation and fertilization were examined for effective IVP. In vitro maturation was carried out using TCM-199 supplemented with 15% FCS and hormones in 5% $CO_2$ incubator for 24h. In vitro fertilization was performed with frozen-thawed sperm in modified mTALP medium containing 0.3% BSA, $10{\mu}g/ml$ heparin, and 5mM/ml caffeine for 24h. The fertilized embryos were co-cultured on monolayer of cumulus cells in TCM-199. When oocytes were collected from functionally active and inactive ovaries, maturation rate was 76.9 and 7.7%, respectively. When oocytes were classified morphologically to good and poor grades, maturation rate was 75 and 58.8%, respectively. FSH + LH + $E_2$ (86.4%) showed higher maturation rate than control (53.0%) and FSH (73%). The fertilization rate was 28.2, 100 and 91.7% in $1.6{\times}10^5$, $5.0{\times}10^5$ and $10.0{\times}10^5$ sperm concentration per ml. When oocytes were fertilized in mTALP and BO media, fertilization and cleavage rates of oocytes in mTALP were higher (84.3 and 56.9%) than those (67.4 and 23.3%) in BO medium. In this experiment, in vitro maturation, fertilization and development of oocytes were affected by type of ovary, grade of oocyte, hormones, sperm concentration and fertilization medium.

• The Korean Journal of Zoology
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• v.39 no.4
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• pp.410-418
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• 1996

Gangilosides are ubiquitous membrane components in mammalian cells and are suggested to play essential roles in cellular phenomena such as cell-cell interadion, differentiation, and signal transduction. Rat ovary contained GM3 as major gangiloside. Nn order to study GM3 distribution in the atretic follicles and its possible changes during follicular development, frozen sections were stained with spedfic monocional antibodies against eleven gangilo-series gangliosides including GM3. In the atretic follicles, Glf3 was expressed in a spatlo-temporally different manner during foilicular development, but GM1 and other gangliosides were not immunohistochemicaily detected. in atretic follicle from primary follicle stage, GM3 was expressed in all the theca cells and some granulosa cells adjacent to oocyte. In atretic follicle from secondary follicle stage, GM3 were expressed in all theca cells and granulosa cells. in atretic follicles from developing Graaflan follicle stages, GM3 was similarly expressed as in secondary follicle stage.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.249-249
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• 2004

This study was conducted to investigate optimal insemination timming as a scanning changes of follicular development by synchronization of ovulation(Ov-synch.) treatment for timed artificial insemination of deer. Sixty-nine elk does were inserted CIDR into virginia for 14 days from 16 to 29 September(breeding season). (omitted)

• Journal of Embryo Transfer
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• v.23 no.2
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• pp.93-100
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• 2008

This study was conducted to establish an in vitro maturation (IVM) system by selection of efficient macromolecule in the porcine in vitro production (IVP) technology. To choose the efficient macromolecules in the development of porcine embryos, the effects of 3 kinds of macromolecules (porcine serum; PS, porcine follicular fluid; pFF, and polyvinyl alcohol; PVA) supplemented in IVM media on the maturation, cleavage, and development rates to blastocyst of parthenogenetic activation (PA) and in vitro fertilization (IVF) embryos were examined. The maturation rates of porcine oocytes in media supplemented with PS were significantly higher than those with pFF and PVA (92.4% vs. 85.4%, 77.1%; p<0.05). In the cleavage and development to blastocyst rates, supplement with PS or pFF in the IVM media was more effective than PA. However, there were no significant differences in cleavage and development to blastocyst between PS and pFF group. From the results of this study, it was demonstrated that PS was optimal macromolecule in the porcine IVM media.