• Clinical and Experimental Reproductive Medicine
• /
• 제18권2호
• /
• pp.123-131
• /
• 1991

In order to study the growth and maturation of ovarian follicle, the localization and activity of alkaline phosphatase(ALPase) and adenosine triphosphatase(ATPase) of the granulosa cells and theca layer were examined according to the follicle size, the follicle state and the ovarian cyclic phase in pig. Theca interna of the small follicles was more heavyly localized with reaction product by the activites of ALPase and ATPase than that of the large follicles. It is assumed that, as the follicles proceed to growth and maturation, antrum formation is the result of the follicular fluid accumulation by means of active transport by the activities of ALPase and ATPase in theca interna. The activities of ALPase and ATPase in atretic follicles were higher than those of normal follicles. These results imply that the mechanisms of follicle maturation and atresia are different according to the phase of ovarian cycle.

• 한국가축번식학회지
• /
• 제17권4호
• /
• pp.279-285
• /
• 1994

The objective of the present study was to characterize the cell cycles of granulosa cell populations during follicular maturation in cattle by using flow cytometer. Granulosa cells were isolated from bovine preovulatory antral follicles of F1(>10mm), F2(5~20mm), F3(3~4mm) and F4(1~2mm) diameter and fixed and stained with fluorochromes that selectively bine to DNA. Flow cytometer equipped with a laser excitation system was used to analyze the intensity of fluorescence from stained cells. Forward angle light-scatter(FSC) and 90$^{\circ}$light-scatter(SSC) signals were adopted to measure the size and the granularity of granulosa cells. As a results of FSC/SSC analysis, granulosa cell populations(G1 phase of cell cycle) from each follicle were relatively regular in size and granularity, regardless of follicular size. However, their distribution in granularity was greater than that in size. Most of granulosa cell populations collected from each follicle were distributed in G0/G1, S and G2/M phases. As the follicles approached to ovulation the percentage of cells in the proliferative phases of cell cycle (S and G2/M) decreased significantly, but there was a concomitant increase in the percentage of granulosa cells in G1 phase. Therefore, these data indicate the proportion of main populations to cell cycle of granulosa cells may be changed from proliferative phase to G1 phase during follicular maturation in cattle.

• 한국가축번식학회지
• /
• 제9권2호
• /
• pp.97-104
• /
• 1985

This study was conducted to investigate the effects of unilateral ovariectomy on the weight of the remaining ovary, the change of number of ovarian follicle, number of corpus luteum and serum progesterone level. Sixty Sprague-Dawley female rats, 23$\pm$2 days old, were divided into 2 groups (control and unilaterally ovariectomized goup) with 30 heads per groups. Each group was again subdivided into 6 groups according to 6 experimental periods; Day 4, 8, 12, 16, 20 and 24 after uniteral ovariectomy. Five arts at every 4 day intervals were sacrificed for the measuring of ovarian weight and for quantitative histologic examination of ovary and at the same time, blood samples were taken for the determination of serum progesterone level of radioimmunoassy. The results obtained were as follows: During the experimental periods, a significant hypertrophy occured in the remaining ovary of unilaterally ovariectomized group from day 16 after operation. The average ovarian weight of control group at day 16 was 21.0$\pm$1.7mg, which is samller than that of unilaterally ovariectomized group weighing 50.5$\pm$8.4mg(P<0.01). The ovarian weight of the unilaterally ovariectomized rats at day 20 and day 24 was 75.9$\pm$2.2 mg and 63.3$\pm$7.0 mg, which is heavier than those of control group; 29.1$\pm$2.3 and 26.3$\pm$1.7 mg(P<0.01 and P<0.01). 2. A same degree of ovarian follicle development was observed in the unilaterally ovariectomized group. Following unilateral ovariectomy and there was no change in total number of follicles larger than 130$\mu$ during the period from day 4 till day 24 after operation. 3. Although the size fo ovarian follicle did not significantly change between two groups from day 4 till day 16, the size of vesicular follicle in unilaterally ovariectomized group (406.3$\pm$26.2$\mu$) was significantly greater as compared to that of control group (323.8$\pm$19.3$\mu$)(P<0.05). 4. Corpus luteum in unilaterally ovariectomized and control group began to a, pp.ar from day 16 after operation and then the number of corpus luteum slightly increased. The number of corpus luteum in unilaterally ovariectomized group at day 24 ws remarkably increased (13.7$\pm$1.41) than that of control (5.2$\pm$2.01)(P<0.01). 5. Serum progesterone levels in unilaterally ovariectomized group were slightly higher than those of control but there were no significant difference between treatment groups.

• 한국수정란이식학회지
• /
• 제15권3호
• /
• pp.287-293
• /
• 2000

This study was undertaken to access the effects of collection method, room temperature at oocyte recovery and culture media on the oocyte quality, fertilization and cleavage rates of in vitro matured and fertilized oocytes of Korean native goats. Ovaries obtained from a slaughterhouse were transported to the laboratory and were divided into 2 groups. One group of ovaries was maintained at 30 to 35$^{\circ}C$ of the room temperature and another group was remained at 20 to $25^{\circ}C$ during oocyte recovery. The oocytes were recovered by follicle aspiration, slicing and aspiration+slicing methods from 3 groups of follicles according to size; <2 mm, 2 to 6 mm and >6 mm. The matured oocytes were inseminated with buck epididymal spermatozoa at a concentration of 3~3.5$\times$10$^{6}$ m1 and fertilization was identified when 2 pronuclei were present in the cytoplasm. Although the recovery rate per ovary obtained by the combination of follicle aspiration + slicing(19.6$\pm$2.2) method was higher than aspiration(11.7$\pm$1.1) and slicing(14.8$\pm$1.8) collection, optimal recovery according to oocyte grades resulted form ovarian slicing compared to aspiration or combined methods(P<0.05). However, no significant differences were found in the mean number(2.5$\pm$1.8; 3.3$\pm$3.3; 2.9$\pm$2.4) and the proportion of favorable oocytes(Grades I, II and III) recovered(31.6%, 36.0%, 36.4%,) according to follicle size(<2 mm; 2 to 6 mm; >6 mm). Fertilization rate was 60.0%, 67.7%, 70.6% and 56.4% and the proportion of embryos/zygotes was 11.1%, 7.1%, 5.0% and 2.8% in 20~$25^{\circ}C$/BO, 30~35$^{\circ}C$/BO, 20~$25^{\circ}C$/TALP and 30~35$^{\circ}C$ /groups, respectively.

• Clinical and Experimental Reproductive Medicine
• /
• 제15권1호
• /
• pp.25-34
• /
• 1988

The intrafo1licular echoes of cumulus oophoruses within ovarian follicles were assessed with the use of ultrasound in 86 women taking part in an in vitro fertilization(IVF) or gamete intrafallopian transfer(GIFT) program, stimulated with pure follicle-stimulating hormone(FSH)/human menopausal gonadotropin(hMG)/human chorionic gonadotropin (hCG). When intrafo1licular echoes were clearly separated from the follicular wall or relatively dispersed within the follicle, they were considered to be a dissociated cumulus, and when they were only slightly prominent from the follicular wall, they were suspected to be a nondissociated cumulus. A cumulus was seen in 62.1% of the follicles larger than 10 mm diameter and 75.1% of them were dissociated. The larger the follicles in size, the more the cumuluses in number and dissociation. The number of follicles and intrafollicular echoes per woman was not different whether or not she would be pregnant, but the number of dissociated cumuluses was significantly more in pregnant women. The number of observed dissociated cumuluses correlated significantly with the number of recovered mature oocytes. When an intrafollicular echo is seen, it can be taken as evidence of a sign of maturity of that particular follicle and oocyte. Ultrasonographic monitoring of intrafollicular echoes and follicular size is very helpful to predict follicular maturation in ovulation stimulation cycles.

• Asian-Australasian Journal of Animal Sciences
• /
• 제19권12호
• /
• pp.1711-1715
• /
• 2006

To predict the number of preantral (primordial, primary and secondary) follicles retrieved from bovine ovaries, we examined the relationship between morphological parameters of ovaries and number of preantral follicles retrieved mechanically. The preantral follicles were retrieved mechanically by slicing ovarian tissue and the influences of size of the ovaries, number of antral follicles, and presence of cystic follicle and corpus luteum on the retrieval were evaluated. Total 77 ovaries were used and significant (p<0.05) relationship was detected between the number of antral follicles and the presence of cystic follicles, and the retrieval number. More preantral follicles were retrieved from the ovaries having more than 20 antral follicles than those having less than 20 antral follicles (17,760${\pm}$5,637 vs. 3,689${\pm}$537) in the ovarian cortex. The retrieval number was significantly reduced in cystic ovaries compared with non-cystic ovaries (5,167${\pm}$825 vs. 20,631${\pm}$6,507). However, neither ovary size (<3.5, 3.5 to 4.0, 4.0 to 4.5 and >4.5 cm) nor the presence of corpus luteum affected the follicle retrieval. In conclusion, the number of preantral follicles retrieved from the ovaries can simply be predicted by the number of antral follicles and the presence of cystic follicles in the ovarian cortex.

• 한국수정란이식학회지
• /
• 제11권2호
• /
• pp.103-109
• /
• 1996

The aim of this study was to evaluate the effect of recombinant bovine somatotropin (bST; Boostin-S, LG Chem) treatment with FSH (Super OV) or PMSG on superovulatory response for transvaginal ultrasound-guided oocyte retrieval (TVR) in calves. Eight Korean Native Cattle(KNC) heifer calves; 150 to 240 days old; were randomly assigned to four treatment groups: 1) FSH(75 mg); 2) FSH (75 mg) + bST(500mg) 3) PMSG(1;000 IU); 4) PMSG(1, 000 IU) + bST(500 mg). Experimental calves in group 1 (n=2) and 2(n=2) were weekly superovulated for 4 consecutive weeks with daily injection of FSH for 3days and the next day subjected to TVR session. Animals in group 3 (n=2) and 4(n=2) were weekly stimulated for 4 consecutive weeks with a single dose of 1, 000 IU PMSG. TVR was performed on 72 hours after PMSG injection. Calves in group 2 and 4 was received injection of 500 mg of bST every 10 days. At each TVR session, follicle number and size were recorded; the oocytes collected and graded according to cumulus and cytoplasm investment. Collected oocyte were determined viable oocyte according to morphological quality with granulation of oocyte and number and status of cumlulus cells. IVM and IVF were performed and assessed cleavage rate on day 3 after fertilization. A Sonovet 600(Medison, Co., Ltd) realtime ultrasound scanner with a 6.5 MHz convex transducer, fixed at the tip of 500 mm estended handle equipped with a needle guide was used in collecting oocyte. Differences between groups were analysed by chi-square test. The population of large follicle ($\geq$5 nun) and aspiration rate were not significant different among the 4 groups. But, the number of small follicles (<5 mm) and aspirated oocyte in the KNC calves treated with bST were 1.3~1.6 times higher than in KNC calves treat with FSH or PMSG alone. In conclusion, the administration of bST with FSH or PMSG at superovulation for TVR in calves was increase the nurnber of small follicle which was influenced the number of aspiratable follicle.

• Clinical and Experimental Reproductive Medicine
• /
• 제13권1호
• /
• pp.39-51
• /
• 1986

This experiment has been done to evaluate the relationship between the follicular atresia and the protein patterns on electrophoreais of the follicular fluids in porcine ovary. The protein concentration of the follicular fluids was lower than that of serum, and gradually decreased as the follicle siae became larger. The number of protein bands of follicular fluid on electrophoresis was less than that of serum, and gradually increased as follicle size became larger. Three specific bands were detected on disc PAGE and one band(M W. 75,000) on SDS PAGE in the follicular fluids, while not in serum. One band (A) at ${\beta}$-globulin region on disc PAGE became heavier, as follicles became atretic. Two bands less than(M. W. 20,000) were detected only in the large follicular fluid. Another band(M. W. 43,000) was not detected in necrotic group, whereas all other groups showed it. It could be concluded that the component and composition of the proteins follicular fluids changes according to the follicular size during atresia. Therefore detection of the changing pattern of proteins in the follicular fluid can be used as a basic criterion for the identification of follicular atretic stage.

• Applied Microscopy
• /
• 제24권1호
• /
• pp.114-122
• /
• 1994

Drastic changes were observed in egg chamber during oogenesis of Aedes togoi immediately after blood meal. The egg chamber, in which the oocyte remains at previtellogenic stage before blood meal, shows very little change in size even for 3 days after emergence but increased 25 folds in volume within 60 hours after blood meal, presumably due to rapid yolk formation. Upto 6 hours after blood meal structures considered to be the precursor of the yolk granules were not observed in the space between follicle cells and oocyte. Vitelline bodies, which are similar to dense bodies secreted from follicle cells, appeared in the space at 10 hours after blood meal. Although vitelline bodies were fused to form vitelline layer, these bodies seem to contribute to the formation of yolk granules. Nurse cells are connected to oocytes by cytoplasmic bridge before blood meal, but the cells are absorbed into oocyte at 6 hours after blood meal.

• Applied Microscopy
• /
• 제28권3호
• /
• pp.263-271
• /
• 1998

Oocyte maturation of the swordtail (Kiphophorus hellerii) was investigated by light and electron microscopy. In the ovary of the swordtail, various staged oocytes were observed, Mature oocytes were located in ovarian cortex, meanwhile immature ones were positioned in ovarian medulla. The oocyte was surrounded by several structures or cells such as chorion, follicle cells, follicular theaca and ovarian epithelium, respectively, from the inside toward outside. Growing and maturing oocytes healed numerous microvilli which interconnected the oocyte and the follicle cells to communicate each other. The mature oocyte had the electron dense chorion which appeared to be ultrastructure of two layers and contained pore canals. Oocyte maturation was characterized by not only the enlarged cell size and well differentiated cell organelles, brit also the increases of fat droplets, pinocytotic vesicles and yolk granules.