• 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.

• Applied Microscopy
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• v.31 no.1
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• pp.19-35
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• 2001

Outlines for plasma $estradiol-17\beta$, components, electrophoretic patterns, and ultrastructural changes were obtained in female rainbow trout (Oncorhynchus mykiss) during the seasonal reproductive cycles. Plasma $estradiol-17\beta$ under the natural conditions, exhibited distinct seasonal variation, peaking very late in vitellogenic season during September, decreasing gradually the halt of spawning in December, and ultimately falling during the early stages of seasonal ovarian recrudescence in February and March. This change in $estradiol-17\beta$ appeared to stimulate vitellogenin production as evidenced by increases in plasma calcium, phosphorus, glucose, albumin and total protein levels. The electrophoretic patterns of late maturing or spawning oocytes were stained more intensively than those of late perinucleolus oocytes (molecular weights of approximately 70,000 and 200,000). Two protein bands were found in the SDS-PAGE separation, coincident with the $estradiol-17\beta$ hormone peak. Gonadosomatic indices (GSI) significantly increased from October to January, and showed the highest peak in January, coinciding with the numerically abrupt increase of ripe ova in female. A positive correlation (r=0.701, p<0.01) was established between plasma $estradiol-17\beta$ levels and the gonadosomatic index during the prespawning. The highest level of hepatosomatic index (HSI) observed in December. During the breeding season (December), the gonadotropes were large and filled with GTH-containing inclusions such as granules and globules. The vitellogenic phase began as late perinurleolus oocytes became transformed into early maturing oocytes through the accumulation of yolk, and oocytes reached the late maturing stages as the ooplasm was completely packed with yolk. Marked ultrastructural changed in the granulosa cells during nuclear migration involve the dilation of the rough endoplasmic reticulum and the appearance of the rod-shaped mitochondria with tubular cristae. Microvilli (finger-like projections), from the zona radiata and from the oocyte grew, and made contact with each other in the pore canals of the zona radials during vitellogenesis, but were withdrawn as the zona radiata became more compact and devoid of pore canals during oocyte maturation. The zona radiata grew to a tripartite structure such as an outer thin homogeneous layer, and two inner thick helicoidal layers (zona radials interna and zona radiata externa). Under the normal conditions, the ovarian follicle influenced the histological development and periodical secretion of the hormones , sufficient for a oogenesis and gonadal steroid production.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.10 no.2
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• pp.71-96
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• 1977

Early development Linuparus trigonus(von Siebold) has been studied based on the samples collected monthly in Je-ju Island, Korea from February, 1975 to January, 1977. Gametogenesis, reproductive cycle, embryonic development were investigated by histological mettled, and morphological description was made on the first phyllosoma larva which reared in the laboratory. Testis is composed of two tubular duct which are symmetrical with H-shaped appearance. Outer layer of testis is of fibrous connective tissue capsule. In the lumen there is a convoluted seminiferous tubule with interstitial tissue. Ovary is a pair of symmetrical blind tubular lobes, and the midportions are connected each other. The ovary consists of a couple of ovarian sacs partitioned by two-layered connective tissue fibers. Proliferation of spermatogonia are observed all the year around on the germinal epithelium of seminiferous tubule. Partial spermatogenesis is always in progress, and the spermatozoa appear all the year around in the tubules. Nutrition of early oogonia is supplied by fibrous mesenchyme which is abundantly distributed in ovarian sacs. Oocytes grow and couplete maturation divisions in the follicle layers. They finally develop into mature ova before spawning. Reproductive cycle is classified into four successive stages; multiplication stage from September to December, growing stage from January to March, maturation division stage from April to May and mature stage from June to August. Spawning takes place from May to August with peak spawning from Into July to early August. Cleavage type is superficial. Blastopore is formed in blasto-disc region which is proliferation of blastoderm cells. Germinal layers are also derived from tile region. Mesoderm formation is originated from endodermal cells which are formed front the blasto-disc region. The endodermal cells are separated by the process of delamination from yolk sac and take part in the formation of the mid-gut. Morphological characteristics of first phyllosoma larva are different from the larvae of other Palinurid and Scyllarid species.

• The Korean Journal of Zoology
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• v.32 no.4
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• pp.337-350
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• 1989

In this study, we compared patterns of hemolvmph with ovarian proteins during the yolk formation of Gewis paludum by using SDS/polyacrylamide gel electrophoresis and he dimen-tlonal electrophoresis. In addition, we examined patterns of glycoprotein which were composed of yolk substances. The results were as follows: The protein patterns in ovary of the 5th instar without eggs were similar to those of adult after ovulation. Protein amounts of the ovaw without developing eggs were less than those of the ovaw containing oocvtes or matured eggs at the molecular weights range from 66, 000 to 110.000 daltons. No glvcoproteins were observed in ovaw without eggs. But the glvcoproteins were gradually increased according to development of eggs in the 5th instar. After the ovulated ovaw of adult, no glvcoprotein bands urere occurred as the bands of the ovary without eggs in the 5th instar. Also, amounts of hemolvmph protein between 66, 000 and 110, 000 in molecular weight were increased during yolk formation of the 5th instal. The results suggest that ovarian protein substances may originate from hemolpnph. In the 5th instar, the amounts of glycoprotein of developing eggs was gradually increased in the hemolymph. The band of M.W. 29, 500 was occurred in the hemolpnph of the 5th instar and adult without eggs. This protein mal be precursor of glvcoprotein in the high molecular weight area in the ovary of more developed eggs. The number of spots and the amounts of protein in ovary without eggs were less than those of ovary containing eggs by two dimensional elec-trophoresis. The protein bands between 45, 000 and 110, 000 almost appeared in acidic field of he dimensional gel. Especially, the band, M.W. 109, 000, uras separated 3 spots, a1, a2, and as. The band, M.W. 102, 000, has spots which designated b1, b2, b3, and b4. Besides, proteins below M.W. 24, 000 were occurred less spots in the basic field than those of acidic field. The mechanism of intracellular organs (= cell organells) uras related to the yolk protein synthesis of oocyte in the process of yolk protein formation of derris pcfudum.

• Development and Reproduction
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• v.8 no.1
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• pp.57-64
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• 2004

This study was carried out to examine the expression of the circadian clock genes in the mouse ovary and testis at different developmental stages. Expression of Period1(Per 1), Period2(Per2), Period3(Per3), Cryptochrome1(Cry1), Cyptochrome2(Cry2), Clock Small and Prokineticin1 and Prokineticin2 receptor(Prok1r, Prok2r) genes in mouse ovary was explored by semiquantitative reverse transcription Polymerase chain reaction(RT-PCR) according to the developmental stage(post partum day; ppd 1, 7, 10, 21 and 35). Immunohistochemistry using PER1 antibody was also analyzed. The differential expression pattern of clock genes was presented according to stages of the mouse ovarian development (ppd 1, 7, 10, 21 and 35). In the cases of ovaries, at the starting point of follicle growth at ppd 7 and 10, the clock gene expression patterns were changed vastly. According to the developmental stages, the clock genes were highly expressed at ppd 7 and 10 in mouse testis also. Receptors for Prok2, the circadian output molecule of SCN, were also expressed in ovary at ppd 7 and in testis at ppd 1 and 7, respectively. Immnunohistochemical analysis of PER1 showed positive signals in the cytoplasm of oocytes and granulosa cells. The level or PER1 expression was increased in cells at the spermatogonia and the condensing spermatids. The expression pattern of Perl and localization of PER1 were showed similar patterns according to the developmental stages in ovary and testis. Taken together, it could be observed that the expression of clock genes was highly correlated with gonadal development and germ cell differentiation in mice. Therefore, in this study, circadian programming of the genes in the ovary and testis is strongly imposed across a wide range of core reproductive cycles and normal development of gametes. Although the existence of circadian genes is clearly investigated, further studies on the direct evidence is required for the understanding of the relationship between circadian genes and regulation of gonadal differentiation and germ cell development.

• Journal of Aquaculture
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• v.4 no.2
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• pp.97-110
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• 1991

This study was undertaken to understand the seasonal changes of hormone levels such as luteinizing hormone (LH), follicle stimulating hormone (FSH) and estradiol-$17{\beta}$, serum components such as albumin, total protein and triglycerides, and GSI of rainbow trout (Onco-rhynchus mykiss) under the normal-light circumstances. LH levels were the highest in November whereas FSH levels were decreased progressively from October to February. Serum GTH, such as LH and FSH, levels were low immediately after egg-stripping and were significantly increased in the summer in association with the initiation of vitellogenesis. Serum albumin, total protein, and triglycerides were significantly increased in August in association with initiation of vitellogenesis and were peak in November prior to egg-stripping. Oocyte growth, measured by oocyte diameter and GSI, was significantly increased in August and showed the highest value in January. Correlation coefficients of serum LH with FSH and estradiol-$17{\beta}$ were +0.844 and +0.947, respectively. Correlation coefficients of estradiol-$17{\beta}$ with albumin and total protein were +0.634 and +0.859, respectively. Correlation coefficients of serum estradiol-$17{\beta}$ with triglycerides and GSI were +0.673 and +0.694, respectively. Probably by positive feedback mechanism, LH, FSH and estradiol-$17{\beta}$ appeared to stimulate to liver to secrete albumin, total protein and triglycerides for vitellosenesis.

• Applied Microscopy
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• v.41 no.2
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• pp.109-116
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• 2011

Polycystic ovary syndrome (PCOS) is hormonal imbalance condition as the endocrine and metabolic disorder that induces the infertility and various complications in reproductive age women. Estradiol valerate (EV) is used hormone replacement therapy in menopausal women and is reported that excessive administration of EV induces the PCOS. Nerve growth factor (NGF) is the factor to regulate the survival and maturation of developing neuronal cell and is also synthesized in ovary. And NGF is overexpressed in EV-induced polycystic ovary (PCO) as previously reported. Therefore, this study examined the possibility of NGF as can be used the biological marker in diagnosis of PCOS, the hormonal imbalance condition, using PCO and CHO (chinese hamster ovarian) cell lines. The concentration of EV treatment is optimized a 1 mg as not influence on the proliferation of CHO cell but 2 mg and 3 mg of EV treatment have the inhibition effect at initial stage. The morphological change was not observed in CHO cell after dose dependent manner treatment of EV. Expression of NGF mRNA and protein is significantly increased at 30 min after EV treatment in CHO cells compared to that of control. And NGF protein expression is strongly increased in PCO tissue, which observed many follicular cysts compared to normal ovary tissue. Taken together, overexpression of NGF may be act as a molecule to induce an abnormal development of follicle, suggesting that NGF can be used as a biological marker in diagnosis of PCOS.

• Clinical and Experimental Pediatrics
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• v.53 no.3
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• pp.420-427
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• 2010

Purpose : Several complications can occur in patients who received bone marrow transplantation (BMT) during childhood and adolescence. This study aims to investigate endocrine dysfunctions after BMT so that better care can be provided to care for long-term survivors of BMT. Methods : One hundred patients (61 males, 39 females) were included in this study. Clinical parameters such as initial diagnosis, age at BMT, conditioning regimen, presence of graft-versus-host disease (GVHD), growth pattern, thyroid function, and pubertal status were retrospectively reviewed to evaluate risk factors associated with endocrine dysfunction. Results : Height standard deviation score (SDS) at BMT, after 1 year of BMT, and at the last visit were $0.08{\pm}1.04$, $-0.09{\pm}1.02$, and $-0.27{\pm}1.18$, respectively (P =0.001). Height SDS significantly decreased in patients who received total body irradiation (TBI) (P =0.017). One of the patients who received TBI demonstrated growth hormone deficiency. Thirty (31.9%) of 94 patients had compensated hypothyroidism. Incidence of compensated hypothyroidism was higher among those who had GVHD (odds ratio 2.82, P =0.025). Of the 32 patients (17 males, 15 females) who were over 14 years in male and 13 years in female at the last visit, 16 (3 males, 13 females) had increased luteinizing hormone (LH) or follicle-stimulating hormone (FSH). Abnormal elevation of LH or FSH was more common in females (odds ratio 30.3, P =0.001). Conclusion : The most common endocrine dysfunction was ovarian insufficiency. Regular check-up for endocrine function needs to be required due to high incidence of endocrine dysfunction in patients with BMT.