• Animal cells and systems
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• 제13권1호
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• pp.49-57
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• 2009

For the study of the reproductive mechanism associated with the process of vitellogenesis, oocyte development and vitellogenesis during oogenesis in female Boleophthalmus pectinirostris were investigated by electron microscopic observations. The ovary consists of a pair of saccular structures with many ovarian lobules. In the early vitellogenic oocyte, the Golgi complex plays an important role leading to the formation of yolk vesicles containing carbohydrate yolks. At this time many pinocytotic vesicles containing yolk precursors are observed in the cytoplasm near the region of initial formation of the zona radiata. In the late vitellogenic oocytes, the multivesicular bodies, which are formed by modified mitochondria, are involved in the formation of the primary yolk granules. Precursors of yolk granules and multivesicular bodies develop to primary yolk globules with participation of pinocytotic vesicles. After primary yolk globules mix with each other, they develop into secondary and tertiary yolk globules. Based on these findings, vitellogenesis of B. pectinirostris occurs by way of the processes of endogenous autosynthesis and exogenous heterosynthesis. The process of autosynthesis involves the combined activity of the Golgi complex, mitochondria, and multivesicular bodies. However, the process of heterosynthesis involves pinocytotic incorporation of extraovarian precursors into the zona radiata of vitellogenic oocytes by way of the thecal cell layers and granulosa cells.

• 한국수의병리학회지
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• 제1권1호
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• pp.72-76
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• 1997

Histological investigation of the number of follicles following gonadotropin treatments for superovulation was carried out in mature Sprague-Dwaley(SD) rats. Routinely serial sections of paraffin-embedded ovaries were stained with hematoxylin-eosin and evaluated with light microscope. During the study unusual cases of microscopic alterations were observed in the medulla of ovaries in two rats. Case one: An ovum and its follicular fluid outflowed in medulla of ovary. The follicular fluid was densly proteinuous. Corona raiata consisted of 2-6 layers thick cells in the periphery of the ovum. While the cortical side of the follicular wall was intact with normal granulosa cell layer the meullary side of it was ruptured. Case two: A large cyst was present in medulla of ovary hilus. The cyst occupied the entire medulla displacing the ovarian archetecture and enclosed by connective tissue and smooth muscle wall.

• Clinical and Experimental Reproductive Medicine
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• 제25권1호
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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.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• 제33권6호
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• pp.879-887
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• 2020

Objective: Numerous studies have indicated that the apoptosis and proliferation of granulosa cells (GCs) are closely related to the normal growth and development of follicles and ovaries. Previous evidence has suggested that miR-126-3p might get involved in the apoptosis and proliferation of GCs, and phosphatidylinositol 3-kinase regulatory subunit 2 (PIK3R2) gene has been predicted as one target of miR-126-3p. However, the molecular regulation of miR-126-3p on PIK3R2 and the effects of PIK3R2 on porcine GCs apoptosis and proliferation remain virtually unexplored. Methods: In this study, using porcine GCs as a cellular model, luciferase report assay, mutation and deletion were applied to verify the targeting relationship between miR-126-3p and PIK3R2. Annexin-V/PI staining and 5-ethynyl-2'-deoxyuridine assay were applied to explore the effect of PIK3R2 on GCs apoptosis and proliferation, respectively. Real-time quantitative polymerase chain reaction and Western Blot were applied to explore the regulation of miR-126-3p on PIK3R2 expression. Results: We found that miR-126-3p targeted at PIK3R2 and inhibited its mRNA and protein expression. Knockdown of PIK3R2 significantly inhibited the apoptosis and promoted the proliferation of porcine GCs, and significantly down-regulated the mRNA expression of several key genes of PI3K pathway such as insulin-like growth factor 1 receptor (IGF1R), insulin receptor (INSR), pyruvate dehydrogenase kinase 1 (PDK1), and serine/threonine kinase 1 (AKT1). Conclusion: MiR-126-3p might target and inhibit the mRNA and protein expressions of PIK3R2, thereby inhibiting GC apoptosis and promoting GC proliferation by down-regulating several key genes of the PI3K pathway, IGF1R, INSR, PDK1, and AKT1. These findings would provide great insight into further exploring the molecular regulation of miR-126-3p and PIK3R2 on the functions of GCs during the folliculogenesis in female mammals.

• 한국동물학회지
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• 제36권2호
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• pp.304-318
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• 1993

1991년 8월부터 그이듬해 7월까지 채취된 무지개 송어(Oncorhvnchus mykiu) 난모세포의발달 단계를 이해하기 위하여 광필 및 전자현미경을 이용하여 본 각구를 실시하였다. 건국 대학교 축산대학 양수.장에서 수온이 14-16$^{\circ}C$이고 용존산소량이 7.6 $\pm$ 0.3ppm. PH 5.8 $\pm$ 0.2인 자연채광 상태인 1개의 10m3 탱크에서 N R.C. 사양표준에 준한 펠렛트 배합사료를 급여하여 연중 사육중인 체중 700-1,2009의 한복새끼인 무지개 송어 암컷 120마리를 공시희.로 이용 하였다. LPO(late perinucleolus oocyte), EMO(early maturing oocyte) 단계에서 과립막 세포와 협막세포와 같은 난포막 세포는 단층구조로 부터 입 방형구조로 변화되었고. 완전한 성장, 성숙 및 과숙기에 방사대(Eons rsdiat3)에 있는 pore canals의 입구가 봉쇄되었다. 방사대는 맨바깥목에 균일하면서 점은 두께의 막과 나선형의 두꺼운 복의 2개의 막으로 구성되어 있으며. 성숙기에 있는 난모세포에는 수초 및 자갈에 붙을 수 있는 온수성 어종 보다 상대적으로 얇은 점액성의 막이 그 주위를 둘러싸고 있다. 난모세포가 성장함에 따라 세포질 주변부 및 임립등 세포내에 막대기 형태의 미토콘드리아와 비대해진 내형질세강의 육가 급증하였다. 전자밀도가 높은 소포는 방사대에 인접한 난세포질 주변부에 많이 존재하며. 이는 성장중인 방사대의 pore Canals을 통해서 간으로 부터 합성된 단백질과 탄수화물을 수송하는 것과 밀접한 관계가 있는 것으로 사료된다. 자연적인 조건하에서 fP포막은 난형성 및 이에 영향을 미치는 성호르몬의 발달에 충분한 대르몬을 계절에 따라 분비시키는 데에 커다란 영향을 미치는 것으로 나타났다.

• Clinical and Experimental Reproductive Medicine
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• 제16권1호
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• pp.23-34
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• 1989

Estradiol (E)은 난소내 과립세포 (granulosa cell, GC)를 증가시키고 생식소 자극호르몬과 협동으로 배란을 유도한다. Androgen은 E의 작용과 반대의 작용을 나타내며 여포의 폐쇄요인으로 알려지고 있다. Testosterone(T)이 폐쇄여포의 여포액내 다량 존재하는 것이 알려짐에 따라 난소내에도 자가조절분비(autocrine)또는 paracrine regulation에 의해 작용을 나타낼 것으로 가정되어 난소내 여포가 폐쇄됨에 따라 그 수용체의 변화하는 양상을 조사하고져 하였다. 흰쥐의 과립세포의 세포질에는 $51.3{\pm}6.1$fmol/mg protein의 Estradiol 수용체(ER) ; $153.1{\pm}25.3$의 Testosterone수용체(TR) ; 또한 $35.1{\pm}8.1$의 Progesterone수용체(PR)가 존재하였다. 과립세포내 ER용 세포질내 E를 제거한 후에 정량이 가능하였고 또한 과립세포내에도 TR이 사람에서는 $23.4{\pm}7.2$ fmol/mg protein, 돼지는 $98.5{\pm}23.1$로 상당량 존재함을 관찰하였다. Dihydrotestosterone Enanthate(DHTE)를 100ug/흰쥐의 농도로 처리한 결과 난소내 TR의 농도는 변화가 없이 ER의 농도만 현저히 저하되고 쥐의 난소무게 역시 감소하는 것을 발견하였다. 위의 결과로 보아 난소내에도 스테로이드 호르몬은 autocrine(자가조절)방법으로 작용하며 An-drogen이 난소의 무게를 감소시키는 것은 ER의 수를 감소시켜 E의 작용이 억제되고 여포들이 폐쇄를 일으켜 그 증식이 저하된 때문으로 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권4호
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• pp.2271-2275
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• 2016

Background: The value of 5-aminolevulinic acid (ALA) in fluorescence detection of peritoneal metastases and the underlying mechanisms were evaluated in patients with peritoneal surface malignancies. Materials and Methods: Oral 5-ALA was administered at a concentration of 20 mg/kg body weight with 50 ml of water 2 hours prior to surgery (n=115). The diagnostic value of 5-ALA based fluorescence production was evaluated following white light inspection during prior to cytoreductive surgery and hyperthermic intraperitoneal chemotherapy. Then, peptide transporter PEPT1 (ALA influx transporter) and ATP-binding cassette transporter ABCG2 (porphyrin efflux transporter) gene expression was determined with quantitative real time (qRT)-PCR and pathological diagnoses confirmed for all tissue samples. Results: The 5-ALA based photodynamic detection rate was 17% for appendiceal mucinous neoplasms, 54% for colorectal cancers, 33% for gastric cancers, 67% for diffuse malign peritoneal mesotheliomas, and 89% for epithelial ovarian cancer of peritoneal metastases. 5-ALA was detected in all cases of peritoneal metastases originating from cholangiocarcinomas whereas it was not able to detect any in granulosa cell and gastrointestinal stromal tumor cases. Furthermore, PEPT1 was overexpressed whereas ABCG2 expression was downregulated in tumors detected with fluorescence. Conclusions: 5-ALA provided 100% specificity and high sensitivity to detect peritoneal metastases in subgroups of patients with peritoneal surface mailgnancies. ALA influx transporter PEPT1 and porphyrin efflux transporter ABCG2 genes are important in tumor specific 5-ALA induced fluorescence in vivo. Further studies should clarify diagnostic utility of 5-ALA in peritoneal surface malignancies.

• The Korean Journal of Physiology and Pharmacology
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• 제14권5호
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• pp.265-272
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• 2010

Follicular cystic ovary (FCO) is one of the most frequently diagnosed ovarian diseases and is a major cause of reproductive failure in mammalian species. However, the mechanism by which FCO is induced remains unclear. Genetic alterations which affect the functioning of many kinds of cells and/or tissues could be present in cystic ovaries. In this study, we performed a comparison analysis of gene expression in order to identify new molecules useful in discrimination of bovine FCO with follicular cystic follicles (FCFs). Normal follicles and FCFs were classified based on their sizes (5 to 10 mm and $\geq25mm$). These follicles had granulosa cell layer and theca interna and the hormone $17{\beta}$-estradiol ($E_2$)/ progesterone ($P_4$) ratio in follicles was greater than one. Perifollicular regions including follicles were used for the preparation of RNA or protein. Differentially expressed genes (DEG) that showed greater than a 2-fold change in expression were screened by the annealing control primer (ACP)-based PCR method using $GeneFishing^{TM}$ DEG kits in bovine normal follicles and FCFs. We identified two DEGs in the FCFs: ribosomal protein L15 (RPL15) and microtubule-associated protein 1B (MAP1B) based on BLAST searches of the NCBI GenBank. Consistent with the ACP analysis, semi-quantitative PCR data and Western blot analyses revealed an up-regulation of RPL15 and a down-regulation of MAP1B in FCFs. These results suggest that RPL15 and MAP1B may be involved in the regulation of pathological processes in bovine FCOs and may help to establish a bovine gene data-base for the discrimination of FCOs from normal ovaries.

• 한국임상수의학회지
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• 제20권3호
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• pp.369-375
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• 2003

The ultrasonography was performed in 77,924 subestrous dairy cows from 680 dairy farms at 57 districts. The cows were examined for confirmation of resumption of ovaries and uterus after 30 days postpartum, for early pregnancy diagnosis after 30 days post-insemination, confirmation of pregnancy at 8 month post-insemination, and for diagnosis of reproductive dysfunctions, respectively. The results of reproductive examination by ultrasonography in 77,924 dairy cows were ovarian disorders 35,452 (46.8%), uterine disorders 10,721 (13.8%), pregnancy and pregnant failure 24,044 (30.8%), adhesion of reproductive tracts 167 (0.2%), freematins 13 (0.01%) and others 6,527 (8.4%) cows, respectively. The 13,732 (49.6%) cows out of 27,694 cows were treated for silent heat or error of estrus detection, 3,473 (12.5%) cows for follicular cysts, 815 (3.0%) cows for luteal cysts, 606 (2.2%) cows for inactive ovaries, 4 (0.01%) cows for granulosa cell tumors, and 6,455 (23.3%) cows for endometritis, respectively. The indices of reproductive efficiencies before (2001-08-10) and after (2003-07-20) periodical examination of reproductive status were as follows; the mean intercalving interval were shortened from 471 days to 383 days, the mean interval calving to conception were shortened from 187 days to 99 days, the mean interval calving to first service were shortened from 105 days to 67 days, the mean cows showing heat by 60 days postpartum were increased from 33% to 89%, the mean conception rate to first service were increased from 43% to 68%, and the mean service per conception were shortened from 2.6 times to 1.8 times, respectively. These results suggest that periodical examination of reproductive status would be useful for the improvement of the reproductive performance in dairy cows.