• 한국동물학회지
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• 제8권2호
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• pp.23-27
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• 1965

The present study is an analysis of some alterations occurring in the population of folliclesin the ovary treated with the gonadotrophins in intact and unilaterally ovariectomized mice. the differences between the findings inthe intact and semispayed animals and those in gonadotrophintreated intact and semispayed animals were disucssed. 1) When mice were semispayed at the stage of diestrus, the remaining ovary showed an increase of secondary follicles in number, while a decrease of atretic folicles, compared with those in intact animals. 2) After injection of PMS and HCG following semispaying at the diestrus stage, the remaining ovary contained larger number of secondary follicles and corpora lutea than those in semispayed animals with no treatment , while showed smaller number of atretic follicles. 3) It is sure that follicular development is remarkably accelerated in the intact mice treated with PMS and HCG. 4) The effect of PMS and HCG on the follicular development was gradually decreased with time in the intact and semispayed animals afte rexogenous hormone therapy. 5) It is believed in the experiment that a decrease of follicular atresia in number and an increase of secondary follicles may be responsible for compensatory reaction after semispaying.

• 한국발생생물학회지:발생과생식
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• 제10권1호
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• pp.1-7
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• 2006

포유류 난포의 폐쇄 과정은 매우 정교한 내분비적 조절작용에 의해 일어나며, 이 과정중에 발생하는 난포 내 과립세포의 퇴화는 핵응축 현상을 동반하는 것으로 알려져 있다. 본 연구는 핵응축 현상과 관련하여 돼지 난소 내 폐쇄난포의 과립세포가 퇴화 시 동반되는 세포 사멸이 아포토시스의 과정에 의해서 일어나는지의 여부와 아포토시스 관련 주요 단백질 분해 효소인 캐스파제-3과 연관된 세포 사멸 기전과 관련이 있는지에 대해서 조사하고자 하였다. 돼지 난소로부터 정상 및 폐쇄난포를 분리하고 이들을 대상으로 조직학적으로 아포토시스 발생 여부를 확인하였고 난포로부터 각각 얻어진 과립세포를 대상으로 하여 아포토시스 과정으로 사멸하는 세포의 형태 및 캐스파제-3의 활성을 관찰 및 측정하였다. 폐쇄 중 돼지 난포 내 과립세포에서 핵의 분절이 흔히 관찰되었고, 유세포 측정기를 이용하여 세포 사멸율을 측정해 본 결과 사멸하는 세포의 수가 폐쇄난포의 과립세포에서 정상난포보다 매우 유의하게 높은 것으로 나타났다. 난포 조직 내 아포토시스 세포는 과립세포에 국한되어 관찰되었으며 협막세포에서는 아포토시스가 관찰되지 않았다. 최종적으로 캐스파제-3의 활성을 정상 및 폐쇄 난포에서 분리한 세포에서 측정해 본 결과 폐쇄난포의 과립세포에서 정상난포보다 유의하게 높은 활성을 보였다. 이와 같은 결과를 종합해 볼 때, 돼지 난포의 폐쇄 중 과립세포의 퇴화는 아포토시스 과정에 의해 일어나며, 캐스파제-3의 활성에 의존적인 것으로 사료된다.

• 대한수의학회지
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• 제37권1호
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• pp.71-78
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• 1997

This study was designed to investigate the number of the growing and mature follicles following gonadotrophin treatments for superovulation in mature rats. Eighteen mature rats (Sprague-Duwely, initially 190~230gm) were randomly alloted into 3 groups. One group was control group, another FSH-treated group was injected intramuscularly with 0.5 units of follicular stimulating hormone (FSH) / rat, and third PMS and HCG-treated group was intramuscularly injected with 20~25IU of pregnant mare serum (PMS) / rat and then at the 48 hrs later, with 20~25IU of human chorionic gonadotrophin (HCG) / rat. The uteri and ovaries of rats were collected and then were observed grossly and serial sections of paraffin embedding ovaries were stained with H-E. Number of ovarian follicles by following 3 grades of large, middle and small follicles from secondary and tertiary follicles were investigated by LM photography of preparations. Small follicles were classified as secondary follicles of preantral follicles with more than 2 layers of granulosa cells surrounding the oocyte and middle follicles were classified as secondary follicles with early signs of antral cavity or with more than one small cavity on either side of the oocytes and large follicles were classified as tertiary follicles with a single medium sized antral cavity or large well-formed antral cavity. In gross findings, the uteri were slightly swelling in FSH-treated group and markedly swelling or filled with fluid in the uterine lumen in PMS and HCG-treated group. In histological findings, the shape and size of the follicles were diverse in middle and large follicles of FSH-treated group and PMS and HCG-treated group, and proportion of atretic follicles was increased in FSH-treated group and PMS and HCG-treated group than those in control group. The uteri of FSH-treated group and PMS and HCG-treated group were hypertropied or filled with fluid in the lumens and walls of uteri. The wall tissue layers were flattened and their blood and lymph vessels were dilated. The mean number of follicle per ovary in control group were appeared to be $17.1{\pm}5.6$($14.0%{\pm}4.6%$), $37.8{\pm}9.1$($30.9{\pm}7.4%$) and $67.6{\pm}30.1$($55.2{\pm}24.6%$) respectively at large, middle and small follicles and total number of these 3 grade follicles were appeared to be $122.5{\pm}40.0$. The mean number of follicle per ovary in FSH-treated group were appeared to be $22.8{\pm}7.0$($17.4%{\pm}5.3%$), $43.4{\pm}6.6$($33.2{\pm}5.1%$) and $64.5{\pm}13.0$($49.3{\pm}9.9%$) respectively at large, middle and small follicles and total number of these 3 grade follicles were appeared to be $130.7{\pm}16.6$. The mean number of follicle per ovary in PMS and HCG-treated group were appeared to be $29.7{\pm}11.0$($16.3%{\pm}6.0%$), $61.9{\pm}17.2$($33.9{\pm}9.4%$) and $91.1{\pm}28.2$($49.9{\pm}15.4%$) respectively at large, middle and small follicles and total number of these 3 grade follicles were appeared to be $182.6{\pm}32.7$. The above findings reveal that large follicles were increased 29.8% in FSH-treated group and 73.7% in PMS and HCG-treated group than those in control group and in histologic findings, proportion of atretic follicles were more increased in ovaries with more number of more developing follicles.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1996년도 공동 춘계학술대회
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• pp.43.1-43
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• 1996

No Abstract, See Full Text

• 한국동물학회:학술대회논문집
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• 한국동물학회 1996년도 공동 춘계학술대회
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• pp.43.2-43
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• 1996

No Abstract, See Full Text

• 한국발생생물학회지:발생과생식
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• 제1권2호
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• pp.157-164
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• 1997

To study the regulation of porcine follicular cell apostosis by gonadotropin, steroid, and nitric oxide, we analyzed DNA fragmentation, the hallmark of apoptosis, and nitrite production of porcine granulosa cells. Dissected indiidual follicles from ovary were separated in size (small, 2-3 mm; medium, 5-6 mm; large, 7-8 mm) and isolated granulosa cells were classified morpholocally as atretic or nonatretic. Nitrite concentration was measured by mixing follicular fluids with an equal volume of Griess reagent. Follicular nitric oxide (NO) concentration of healthy follicles was higher than that of atretic follicles. Apoptotic DNA fragmentation was suppressed in non-apoptotic granulosa cells. Follicular apoptosis was induced by androgen but prevented by gonadotropin in vitro. Apoptosis was confined to the granulosa cells. But it was not clear whether apoptosis of granulosa cells were isolated, incubated with or without gonadotropin, androgen and sodium nitroprusside (SNP), respectively at $37^{\circ}C$ for 24 hrs. Cultured granulosa cells were used to extract genomic DNA and culture media was asssayed for nitrite concentration. Nitrite production of culture media was increased, while apoptotic DNA fragmentation was suppressed in PMSG, hCG, testosterone+SNP and SNP treated groups. Nitrite concentration in culture media was decreased, but apoptotic DNA fragmentation was induced in testosterone treated group. These data suggest that NO production and apoptosis may be involved of granulosa cell apoptosis induced by testosterone.

• Clinical and Experimental Reproductive Medicine
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• 제30권1호
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• pp.47-55
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• 2003

Objectives: Bok, Bcl-2-related ovarian killer, is a proapoptotic Bcl-2 family protein identified in the ovary based on its dimerization with the antiapoptotic protein Mcl-1. The present study examined the hormonal regulation and localization of Bok messenger RNA levels in the mouse ovary during the follicle development. Methods: The animals were implanted subcutaneously with Silastic brand capsules containing the synthetic estrogen, DES at $21{\sim}23$ days of age. Ovaries were collected $1{\sim}3$ days after implantation for RNA analysis and in situ hybridization. Some mice were removed capsule for $1{\sim}2$ days to induce ovarian follicle apoptosis. Ovaries were also collected from 26 day-old immature mice at various times after treatment with 10 IU PMSG. Some mice received a single intraperitoneal injection of 10 IU hCG to induce ovulation, and ovaries were obtained at different time intervals for Northern blot and in situ hybridization analysis, respectively. Results: Treatment of immature mice with diethylstilbestrol (DES) for $24{\sim}48$ h increased ovarian Bok mRNA levels. Bok mRNA was remained the same levels in mice removed DES for $24{\sim}48$ h to induce apoptosis. High signals of Bok mRNA after DES treatment were detected in granulosa cells of early antral follicles. Treatment of immature mice with PMSG for 12 h increased markedly ovarian Bok mRNA expression which was detected mainly in preantral and atretic follicles. Interestingly, low levels of Bok mRNA were also expressed in granulosa cells of preovulatory follicles. Treatment of PMSGprimed mice with hCG stimulated strongly ovarian Bok mRNA expression at $6{\sim}9$ h. At that time, Bok mRNA was expressed in granulosa cells of atretic and small growing follicles. Conclusion: These results demonstrate that Bok is one of proapoptotic Bcl-2 members expressed in early growing and atretic follicles during the ovarian follicular development. Gonadotropins induce a transient increase of Bok gene expression in granulosa cells of preantral and preovulatory follicles indicating some role in the ovulatory process.

• 한국동물생명공학회지
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• 제35권4호
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• pp.329-337
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• 2020

Cytochrome P450 1A2 (CYP1A2) is a member of the cytochrome P450 superfamily enzymes in mammals and plays a major role in metabolizing endogenous hormones in the liver. In recent days, CYP1A2 expression has been found in not only the liver but also other tissues including the pancreas and lung. However, little information is available regarding the expression of CYP1A2 in the ovary, in spite of the facts that the ovarian follicle growth and atresia are tightly associated with controls of endocrine hormonal networks. Therefore, the expression of CYP1A2 in the ovaries of prepubertal and pubertal rats was investigated to assess its expression pattern and puberty-related alteration. It was demonstrated that the expression level of CYP1A2 was significantly (p < 0.01) higher in the pubertal ovaries than prepubertal counterparts. At the ovarian follicle level in both groups, whereas CYP1A2 expression was less detectable in the primordial, primary and secondary follicles, the strongly positive expression of CYP1A2 was localized in the granulosa cell layers in the antral and pre-ovulatory follicles. However, the ratio of CYP1A2-positive ovarian follicle was significantly (p < 0.01) higher in the ovary of pubertal group (73.1 ± 3.1%) than prepubertal one (41.0 ± 10.5%). During the Immunofluorescence, expression of CYP1A2 was mainly localized in Fas-positive follicles, indicating the atretic follicles. In conclusion, these results suggested that CYP1A2 expression was mainly localized at the atretic follicular cells and affected by the onset of puberty. Further study is still necessary but we hypothesize that CYP1A2 expresses in the atretic follicles to metabolize residue of the reproductive hormones. These findings may have important implications for the fields of reproductive biology of animals.

• 한국가축번식학회지
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• 제16권2호
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• pp.165-173
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• 1992

In order to study the mechanism of follicle growth and maturation, and also to supplement the criteria identifying the follicle state of normal of atretic, the histochemical investigation on the ovarian follicles according to the ovarian cycle of mouse, rat and pig has been done. The intercellular space of granulosa cells, especailly Call-Exner body, and follicular fluid in the antrum showed positive to PAS, and blue stain by trichrome dye. The resutls suggest that the mucous polysaccharide was synthesized by the granulosa cells, and secreted into the antrum through Call-Exner body so as to be the components of the follicular fluid as the follicles proceeded to growth and maturation. The further the follicles proceeded to atresia the more densely their theca externa were stained blue by follicles proceeded to atresia the more densely their theca externa were stained blue by trichrome dye, and the more densely the granulosa cells were stained red by oil red 0 dye. Therefore, these staining methods can be applied to the criteria identifying the follicle atresia.

• Nuclear Engineering and Technology
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• 제31권4호
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• pp.385-390
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• 1999

Ovarian follicles are faced with one of two fates, atresia or development. Up to 99% of follicles become degenerated rather than ovulated in female life span. Thus, atresia occurs at all stages of follicle development in mammalian ovaries. In the present experiment, the effect of ${\gamma}$-radiation on primordial follicles was morphologically analyzed in a mouse ovary. Thirty-seven percent of the primordial follicles in the non-irradiated control mice ovaries were abnormal. At day 8 post irradiation, most of primordial follicles became atretic. They lost their integrity of architecture in the follicular shape. Then, all the oocytes disappeared from the follicles. And only 3 to 4 granulosa cells lay down onto the basement membrane. Disappearance of granulosa cells or oocytes resulted from the radiation-induced apoptotic process. It is definitely clear that ${\gamma}$-radiation induces rapid apoptotic degeneration of the primordial follicles. The morphological degeneration induced by radiation in the primordial follicles can be used as an experimental model to draw out a deeper insight for radioprotectant researches.