• Korean Journal of Ichthyology
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• v.13 no.4
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• pp.254-260
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• 2001

In the goby Micropercops swinhonis, the follicular layer of full-grown oocytes consists of an outer layer (theca cell) and an inner layer (granulosa cell). As the oocyte grows, columnar cells of inner granulosa layer secrete mucin to their cytoplasm and then surround the oocyte. Such granulosa cells appear to be cuboidal cells in the early vitellogenesis, yolk vesicle stage, to be replaced by columnar cell secreting mucins (adhesive materials) in the middle vitellogenesis, yolk granule stage. The enveloping layer of the oocyte has a muco- follicle layer filled with mucins. The mucins are an amorphous and electron-dense substance. Interestingly, the oocyte enveloping layer becomes thickened towards the animal pole as vitelogenesis proceeds. A zona radiata of about $7.8{\sim}11.5\;{\mu}m$ thick is present below the muco-follicle layer. The zona radiata is composed of an one-layered electron-dense externa and a three to five-layered electron-less interna.

• Journal of Animal Science and Technology
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• v.46 no.4
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• pp.571-584
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• 2004

It is known widely that granulosa cell apoptosis leads follicular atresia and macrophages exert their effects directly and/or indirectly from the initiation to the completion of follicular atresia by phagocytosis of apoptotic bodies and secretion of various cytokines. However, the site of initiation, propagation routes and the elimination methods of apoptotic bodies, and the time and methods of penetration of macrophages into the follicles are not known completely. Using pig(Yorkshire-breed) ovary, immunohistochemical studies with TUNEL for apoptotic bodies and pig macrophage monoclonal antibody 4E9 for macrophages, and light and transmission electron microscopic observations were performed. In the pig, follicular atresia began with the granulosa cell apoptosis, and the apoptosis of theca intema cells occured at the same time. The apoptosis of granulosa cells initiated randomly within the granulosa cell layer and propagated rapidly into the whole layer. Ultrastructura1ly, apoptotic granulosa cells showed characteristic pyknotic and deformed nucleus and intracytoplasmic vesicles. Apoptotic bodies were eliminated by intact granulosa cells and macrophages. Intact granulosa cells ingested apoptotic bodies transiently, soon after they fell into the apoptosis. Finally, apoptotic bodies and degenerated oocyte were phagocytosed by macrophages. Macrophages entered the ovarian follicle at the time of initiation of granulosa cell apoptosis, and migrated with the progression of apoptosis. By elimination of theca cells, macrophages contributed the completion of follicular atresia These results will provide valuable informations on the study of the interrelation between macrophage and ovarian follicular atresia.

• Korean Journal of Veterinary Research
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• v.27 no.1
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• pp.141-151
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• 1987

A total of 1200 Korean native cow and 240 dairy cow genitalia were collected during the slaughtering process in Seoul and Kwang Ju abattoir and were examined from July 1985 to March 1986. Ovarian follicles were classified as cystic if the diameter was greater than 2.5cm or if follicles were multiple. In order to investigate the ovarian cysts, anatomical and histological examinations were performed. In addition progesterone and estrogen level in different types of cystic follicular fluid and serum were measured by radioimmunoassay. The results were summerized as follows: 1. The incidences of ovarian cysts were 2.0% in Korean native cow and 7.9% in dairy cow. 2. In distribution of cysts in the left, right and both ovaries, the most encountered ovary was right one. The frequency was 45.8% in right ovaries, 33.4% in left ovaries and 20.8% in both ovaries in Korean native cow. On the contrary the frequency was 42.1% in right ovaries, 31.8% in both ovaries and 26.3% in left ovaries in dairy cow. 3. Six speciemens (25.0%) of Korean native cow and six specimens (31.6%) of dairy cow were associated with corpora lutes in both ovaries. 4. The luteinization of theca layer was most significant in the group 2Aa (71.4%) and 2Ba (38.5%) which associated with no granulosa cell and corpora lutea in the same cystic ovaries. 5. Correlation of progesterone concentration between cystic fluid and serum was found only in the group 2Aa and 2Ab (r=0.86). Progesterone and estrogen concentrations in cystic fluid were closely related to the degree of degeneration of granulosa cell layer. The cystic follicles that consist of thickened theca and degenerated granulosa cell layers contained a large amount of progesterone, and small amount of estrogen. In conclusion, various types of ovarian cysts with various levels of progesterone and estrogen were observed in Korean native cow.

• The Korean Journal of Zoology
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• v.39 no.3
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• pp.273-281
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• 1996

Previously, we have proposed a two-cell type model for follicular steroidogenesis inamphibians with Rana nigromacu lota. Present experiments were carried out to ascertain whether the model Is applicable to R. dybowskii. The role of theca layer were also reassessed by using granulosa cell-free pure theca layer (P-THEP). Theca/epithelium (THEP) layers, P-THEP layers, and granulosa cell enclosed-oocytes () were obtained from ovarian follicles of R. dybowskii by microdissection. Intact follicles (IFs) and different types of tissues were cultured for 6 hour in amphibian Ringer's m the presence or absence of FPII (0.05 gland/mi) or various steroid precursor (100 ng/ml). The amounts of product steroids converted by the components were measured by RIA. Exogenously added pregnenolone (P5) resulted in a marked increase in progesterone (P$_4$) by GCEOs (2143 pg/follicle) and IFs (2346 pg/follicle) but a smaller increase in P4 by THEP layer (495 pg/follicle). Addition of P$_4$ increased 17 a-hydroxyprogesterone (17 $\alpha$-OHP$_4$) levels by GCEOs (1118 pg/follicle) and IFs (1333 pg/follicle) but less by THEP layer (290 pg/follicle). However, much less amounts of P$_4$ or 17 $\alpha$-OHP$_4$ were producad by P-THEP layers than THEP in the presence of P5. Exogenous 1 7$\alpha$-OIIP$_4$ increased androstenedione (AD) levels by GCEOs (1415 pg/follicle) and IFs (561 pg/follicle) but not by THEP layers. In contrast, addition of AD resulted m a marked increase in testosterone (T) levels by TIIEP (2594 pg/follicle) and IFs (2223 pg/follide) but much less by GCEOs (339 pg/follicle). Exogenous T increased estradiol (E$_2$) levels by GCEOs (551pg/follicle) and IFs (887 pg/follicle), but not by THEP layer (<10 pg/follicle). Without addition of FPH or steroid precursors, very low or nondetectable levels of steroids were produced (< 20 pg/follicle) by all the types of follicular components examined. The data presented here indicate that the two-cell type model based on the study with R. nigromacu Iota is applicable to R. dybowskii and also suggest that the minor pathway, which convert P5 to 17$\alpha$-OHP$_4$, is not present in theca layer.

• The Korean Journal of Zoology
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• v.38 no.4
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• pp.523-530
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• 1995

Specific affinity binding sites for atrial natriuretic peptide (ANP) were Investigated in the pig ovarian tissues by in vitro autoradiographic techniques. In the pig ovary, the highest binding sites for 12514abeiled rANP(l~28) were localized in the granulosa cell layer of the forncles. The binding sies on theca layer of the ovarian follicles were mainly localized in the external layer, but none was observed In the Internal layer. In the corpus luteum, the binding site was not observed. The specific bindings of 200 pM of l2Sl4abelled rANP(l~28) to granulosa and theca externa layers were reversed completely by excess concentration (1 ~4) of unlabelled rANP(l~28) but not by 10 ~ of unrelated peptides, human angiotensin II and arginine vasopressin. The binding was also displaced by 1 ~ of desiGIn18, Ser19, Gly2O, leu21, Gly22I ANP(4~2g) (C- ANF) as a spedfic ligand of the ANP clearance receptor. Therefore these results indicate ~hat the biological and the clearance ANP receptors exist in the theca externa and granulosa layer of the pig ovary, and suggest that the ANP receptors may be related with the regulatory lundion of the ovarian follicular development including oocyte maturation.

• Korean Journal of Ichthyology
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• v.19 no.2
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• pp.112-119
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• 2007

A histological study on development and transformation of the oocyte' follicle cell for Korean four sillurid fishes, Liobagrus obesus, L. mediadiposalis, Pseudobagrus koreanus, and P. brevicorpus was performed by light and electron microscopes. The follicular layer surrounding the oocyte consisted of an outer theca cell and an inner follicle cell (granulosa cell). The follicle cells of the oocyte were flatten cells at early oocyte but during vitellogenesis they were transformed it to a single layer of cuboidal cell, then to a single columnar cell layer, and finally to a layer covered with a substance secreted by themselves. Although the development and transformation of the follicle cells was similar to four species, the secreted materials, called an adhesive membrane, were divided into two types in its appearance and nature. Firstly, a jelly coat-like type was found in L. obesus and L. mediadiposalis, which they are presumed to be polysaccharides and mucoproteins in its nature and secondly, a granular type in P. koreanus and P. brevicopus, being mucoprotein. A zona radiata with about $0.6{\sim}3.1{\mu}m$ thin was present below the adhesive material secreted by the transformed-follicle cell's activity. The zona radiata was composed of two layers, a thin externa and a thick interna.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.3
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• pp.195-200
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• 2002

Objective : To investigate the expression of apoptosis related proteins and apoptotic cells on the human ovarian follicles. Materials and Methods: Thirty five Formalin-fixed paraffin-embedded human ovarian tissue blocks were selected from the surgical pathology files of the department of pathology, College of Medicine, Yonsei University, for the period from 1996 to 1998. All specimen were from premenopausal women aged from $32{\sim}45$. Ovarian tissues were collected from the patients performing hysterectomy for benign uterine diseases. Immunohistochemical staining was performed for the detection of DNA fragmented cell, Bcl-2, Bax, Fas and Fas-ligand. Results: Bcl-2 and bax were not expressed on the surrounding cells and oocyte of the primary, primordial and preantral follicles. Fas and Fas-ligand (Fas-L) were not expressed on the surrounding cells on the primordial and primary follicles. But expressed on the surrounding granulosa cells and oocyte in the primordial and primary follicles. In the healthy follicles, Bcl-2 was expressed on the granulosa cells, however, Bax was not expressed. DNA fragmented cells were expressed on the inner granulosa cell layer of atretic follicles. Conclusion: Fas, Fas-ligand, and Bax may be responsible for the follicular atresia and Bcl-2 may be involved in the follicular survival in the human ovary.

• Clinical and Experimental Reproductive Medicine
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• v.12 no.1
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• pp.83-98
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• 1985

In order to study the mechanism of follicular atresia, the follicles of the porcine ovary were isolated according to the presence or absence of the corpus luteum and their size, and then classified to the normal? or atretic?follicle on the morphological observation such as the transparency, the vascularization of follicle, the nuclear phase of oocyte, and the homogeneity of the granulosa cell layer. The viability of granulosa cells was examined. The concentrations of progesterone ($P_4$), testosterone (T), and estradiol-17 beta ($E_2$) in each follicular fluid were estimated by the radioimmunoassay. The viability of granulosa cells in the atretic follicle was much lower than that of the normal one. The concentration of each steroid hormone increased as the follicular size was increased, was not different in quantity between the normal- and the atretic follicle of which diameter was below 3mm, and were much higher in the atretic follicle than those in the normal one of which diameter was above 7mm. The ratio of the concentration of E2 to T in the large atretic follicle valued higher than that in the normal one, but smaller in the small and medium atretic follicle than that in the normal one. The present study suggests that the mechanism of atresia of the large follicle may be different from that of the small and the medium follicle and that the amount of steroid hormones regarded as the one of the criteria for the atretic follicles.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.1
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• pp.93-102
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• 1998

Normal and abnormal follicular growth and steroidogenesis depend on gonadotropins as well as intraovarian peptides, which may mediate or potentiate gonadotropin action. Inhibin also affect follicular development and steroidogenesis and may play a role in dominant follicle selection and follicular atresia. Therefore, we studied the differences of serum inhibin, gonadotropin and androgen levels in the women with only the ultrasound findings and no disorder, and polycystic ovary (PCO) with ovulatory disturbance. We prospectively analysed forty-three women with PCO. The diagnosis of PCO was based on typical appearance of the ovaries on TVS. Twelve women with regular menstrual cycle and normal ovarian morphology were selected as control. Basal levels of inhibin, luteinizing hormone (LH), follicle stimulating hormone (FSH), estradiol $(E_2)$, testosterone (T), androstenedione (ADD), dehydroepiandrosterone-sulfate (DS), prolactin and TSH in serum were determined. There were significant differences in basal LH levels and LH/FSH ratio between the control and the women with PCO. The basal levels of inhibin and $E_2$ in the oligo-amenorrheic PCO (N=34) were significantly higher than those in the control. There was higher negative correlation between the inhibin and T levels in the oligo-amenorrheic PCO, but, not in the regular cycling PCO. Also, there was higher positive correlation between the LH and T levels in the oligo-amenorrheic PCO, but not in the regular cycling PCO. These data presume that the initial event of PCO is elevated pituitary LH secretion. Elevated levels of LH may down-regulate LH receptors on granulosa cells and also cause hypertrophy of the thecal layer. High level of androgen secreted by the hypertrophied thecal layer may stimulate inhibin secretion from granulosa cells and can be converted to estrogen by extraovarian tissues and could serve to augment pituitary sensitivity to GnRH with a resultant secretion of more LH than FSH. Inhibin may inhibit FSH action on granulosa cell in the PCO follicle, impairing follicular development and dominant follicle selection resulted in ovulatory disturbance.

• The Korean Journal of Zoology
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• v.35 no.2
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• pp.136-143
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• 1992

돼지 난포의 폐쇄기 작을 규명하고 폐쇄난포의 판정기준을 보완, 확립하기 위하여 난포의 크기 및 상태 그리고 난소주기에 따라 Acid phosphatase(Acpase)의 활성부위와 활성도를 조사하였다. Acpase의 활성부위는 과립세포와 난포막내층이었고, 반응정도는 난포 :극 대난포와 중난포의 경우 정상군에 비해 폐쇄군에서 강한 반응을 보였다. 황체기난포의 과립세포에서는 중난포와 소난포 모두 반응이 미약하였고, 난포막층에서는 정상군과 폐쇄군 모두 강하게 나타났다. Acpase의 활성도는 난포기 난포의 과립세포에서 난포가 커짐에 따라 증가하였는데 난포막층의 활성도는 감소하였으며 , 폐쇄군이 높은 활성도를 보f:다. 황체기의 경우 반포기와는 달리 과립세포와 난포막층에서 난포가 커짐에 따라 활성도는 감소하였으나 폐쇄군은 역시 높았다. 이상의 결과에서 폐쇄난포의 Acpase의 활성부위와 활성도는 정상난포에 비하여 뚜렷하고 강하여서 이 방법은 폐쇄의 판정기준으로 이용 가능한 것으로 사료된다. 또한 난소주기에 따라 상이한 결과를 나타낸 것은 폐쇄기 작이 서로 다름을 시사하였다.