• Journal of Life Science
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• v.19 no.4
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• pp.436-441
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• 2009

B23/nucleophosmin, a nucleolar protein, translocates into the nucleus from the nucleolus when cells are damaged by extracellular stresses. Recently, it was shown that such translocation of B23/nucleophosmin in normal fibroblasts under stress conditions increases both the stability and activation of the p53 protein by disrupting its interaction with MDM2. Senescent cells have a single large nucleolus and a diminished capacity to induce p53 stability upon exposure to various DNA damaging agents. To investigate the role of B23/nucleophosmin in p53 stability in senescent cells, we established a senescence model system by expressing the ras oncogene in IMR90 cells. The stability of p53 was reduced in these cells in response to nucleolar stress, although the level of B23/nucleophosmin protein was not changed. In addition, p53 did not accumulate in the nucleus and B23/nucleophosmin did not translocate into the nucleoplasm. The binding affinity of B23/nucleophosmin with p53 was reduced in senescent cells, whereas the interaction between MDM2 and p53 was stable. Taken together, the stability of p53 in ras-induced senescent cells may be influenced by the ability of B23/nucleophosmin to interact with p53 in response to nucleolar stress.

• Journal of Aquaculture
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• v.18 no.3
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• pp.167-172
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• 2005

Sex reversal to a functional male of sevenband grouper $(41.0{\pm}1.3cm\;TL,\;1.4{\pm}0.1kg\;BW)$ was induced by $17{\alpha}-methyltestosterone\;(MT,\;0.5\sim2.0mg/kg\;BW)$ implantation from March 17 to May 12,2002. Gonad of control group was composed of genial cells and peri-nucleolus oocyte during the experimental period. Gonad of fish treated with 0.5 mg MT/kg BW had peri-nucleolus oocytes, spermatogonia, spermatids and spermatozoa at the late stages of spermatogenesis, while the fish group treated with 1.0 and 2.0 mg MT/kg BW contained spermatoza in the efferent duct. Sperm were obtained from the experimental groups treated with a dose of $1.0{\sim}2.0mg$ MT/kg BW. In the MT treated groups, testosterone and 11-ketotestosterone levels were higher than those in the control group during the $2{\sim}6$ weeks of the experimental period (P<0.05). $Estradiol-17{\beta}$ was detected from fish in the experimental fish.

• Korean Journal of Plant Tissue Culture
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• v.25 no.6
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• pp.453-475
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• 1998

During the 100 years since the initial discovery of meiotic phenomenon many brilliant aspects have been elucidated, but further researches based on light microscopy alone as an experimental tool have been found to have some limits and shortcomings. By the use of electron microscopy and armed with the advanced knowledges on modern genetics and biochemistry it has been possible to applu molecular technology in gaining information on the detailed aspects of meiosis. As synapsis takes place, a three-layered proteinous structure called the synatonemal complex starts to form in the space between the homologous chromosomes. To be more precise, it begins to form along the paired chromosomes early in the prophase I of meiotic division. The mechanism that leads to precise point-by-point pairing between homologous chromocomes division. The mechamism that leads to precise point-by-point pairing between homologous chromosomes remains to be ascertained. Several items of information, however, suggest that chromsome alignment leading to synapsis may be mediated somehow by the nuclear membrane. Pachytene bivalents in eukaryotes are firmly attached to the inner niclear membrane at both termini. This attached begins with unpaired leptotene chromosomes that already have developed a lateral element. Once attached, the loptotene chromosomes begin to synapse. A number of different models have been proposed to account for genetic recombination via exchange between DNA strands following their breakage and subsequent reunion in new arrangement. One of the models accounting for molecular recombination leading to chromatid exchange and chiasma formation was first proposed in 1964 by Holliday, and 30 years later still a modified version of his model is favored. Nicks are made by endomuclease at corresponding sites on one strant of each DNA duplex in nonsister chromatid of a bivalent during prophase 1 of meiosis. The nicked strands loop-out and two strands reassociate into an exchanged arrangement, which is sealed by ligase. The remaining intact strand of each duplex is nicked at a site opposite the cross-over, and the exposed ends are digested by exonuclease action. Considerable progress has been made in recent years in the effort to define the molecular and organization features of the centromere region in the yeast chromosome. Centromere core region of the DNA duplex is flanked by 15 densely packed nucleosomes on ons side and by 3 packed nucleosomes on the other side, that is, 2000 bp on one side and 400 400 bp in the other side. All the telomeres of a given species share a common DNA sequence. Two ends of each chromosome are virtually identical. At the end of each chromosome there exist two kinds of DNA sequence" simple telpmeric sequences and telpmere-associated sequencies. Various studies of telomere replication, function, and behabior are now in progress, all greatly aided by molecular methods. During nuclear division in mitosis as well as in meiosis, the nucleili disappear by the time of metaphase and reappear during nuclear reorganizations in telophase. When telophase begins, small nucleoli form at the NOR of each nucleolar-organizing chromosome, enlarge, and fuse to form one or more large nucleoli. Nucleolus is a special structure attached top a specific nucleolar-organizing region located at a specific site of a particular chromosome. The nucleolus is a vertical factory for the synthesis of rRNAs and the assenbly of ribosome subunit precursors.sors.

• Journal of Aquaculture
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• v.9 no.1
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• pp.73-81
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• 1996

This study was conducted to study the reproductive cycle in female fusilier, Caesio diagramma by using the histological methods. Histological changes of the ovary were well correlated with the patterns of gonadosomatic index (GSI) and hepatosomatic index (HSI). GSI was increased in April as the value of HSI increase and reached to its maximum in May and June. Oocytes at the chromatin-nucleolus and peri-nucleolus stages were observed in the ovary throughout the year. In April, oocytes containing yolk appeared in ovaries of a few fishes. Most oocytes appearing in May and June belonged to the tertiary yolk stage. Frequency of oocytes appearance at the tertiary yolk stage in May and June was higher than that of the other months. Moreover, the empty follicles and atretic oocytes were observed in the ovaries with many vitellogenic oocytes during these two months. Thereafter, oocytes of the yolk stage disappeared in September. The spawning period of C. diagramma is from the month of April to June, and this species belongs to an asynchronous and multiple spawner.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.39 no.3
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• pp.297-302
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• 2006

We investigated gonadal development and sex ratio of artificial seedlings of the oblong rockfish Sebastes oblongus, based on samplings for 370 days just after parturition. The primordial germ cells and genital ridge appeared separately under the mesentery in the yolk-sac stage larva (total length: 7.10-7.77 mm) just after parturition. The primordial germ cells and genital ridge integrated to form primordial gonad in 5-day-old larvae (7.12-9.68 mm), and then proliferation of somatic cell and germ cell occurred in the gonad, which was maintained undifferentiated until 45-days after parturition (18.6-20.4 mm). The ovarian differentiation began in the larva of 50-days old (dab) after parturition (dap) (20.0-24.5 mm). The somatic tissues elongated from the both opposite end-sites of undifferentiated gonad were consequently fused and formed a complete ovarian cavity at 60-days old dap (25.5-32.0 mm). In 80-days old dap (37.3-47.2 mm), meiosis of oogonia occurred to be chromatin nucleolus stage oocyte. The perinucleolus stage oocytes appeared at in 130-days old dap (68.0-86.0 mm), and previtellogenic stage oocytes appeared in 370-days old dap (101.0-116.0 mm). Only female was observed in the artificially produced oblong rockfish in the present study. This result revealed the effect of higher temperature on the sex determination of the oblong rockfish..

• Korean Journal of Fisheries and Aquatic Sciences
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• v.39 no.5
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• pp.404-409
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• 2006

This study investigated the reproductive cycle of the female hairychin goby, Sagamia geneionema, histologically. The fecundity of female hairychin goby ranged from 1,002 to 1,240 eggs when they reached a total length of 9.1-10.0 cm. Fecundity is related to total length. The gonadosomatic index (GSI) increased from December (1.87$\pm$0.46) and reached a maximum in April (11.57$\pm$1.92). The histological changes in the ovary were correlated with the GSI. Oocytes at the chromatin-nucleolus and peri-nucleolus stages were observed in the ovary year-round. In December, oocytes containing yolk appeared in the ovaries of a few fish. Most oocytes appearing in January were at the yolk globule stage. The frequency of oocytes appearing at the yolk globule stage from January to March was higher than in other months. Subsequently, empty follicles and atretic oocytes were observed in the ovaries in May. Based on the histological observations of gonad development and the monthly change in the GSI, the reproductive cycle was classified into the following successive stages: growing (October to November), mature (December to January), ripe and spawning (February to April), and degenerative and resting (May to September) stages. The histological observations of ovaries during the spawning period indicate that this species is a multiple spawner with abbreviated iteroparity based on the developmental pattern of oocytes.

• Korean Journal of Ichthyology
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• v.20 no.4
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• pp.255-262
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• 2008

The endemic Korean stumpy bullhead Pseudobagrus brevicorpus is a first-grade endangered wild fish as designated by the Ministry of Environment of Korea. As part of its restoration and proliferation effort, a histological study of this fish was carried out to investigate sex differentiation and gonadal development based on F1 generation individuals obtained by artificial breeding. On days 4~5 after hatching, a pair of genital ridges including clusters of primordial germ cells was observed between the gut and the mesonephric duct. On days 20 after hatching, the ovary began to initially differentiate and contained early oocytes with chromatin-nucleolus and peri-nucleolus stages on days 30~40 after hatching. As yolk material accumulated after day 80 from hatching, the oocytes grew increasingly large and were surrounded by a distinct follicular layer. On days 306 after hatching, the oocytes grew toward a mature ovum. In the males, the testis was distinguished by emergence of spermatogonium cells on 25 days after hatching, and day 40 after hatching it contained a small number of seminal lobes forming cysts. From 173 days after hatching, the testis consisted of numerous enlarged seminal lobes including spermatocytes and spermatids. Over 14 months after hatching, some seminal lumens were filled with spermatozoa.

• Parasites, Hosts and Diseases
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• v.25 no.2
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• pp.99-109
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• 1987

The present study was carried out to examine the effect of a calcinogen, aflatoxin B1 on the ultrastructural changes of ciliary epithelial cells in mice infected with Clonerchis sinensis. A total of 93 male albino mice(BALB/c strain) was divided into 3 groups; group I, treated with 1.0 ppm aflatoxin Bl for 12 weeks; group II, given 50 C. sinensis n;etacercariae, and group III, given 50 metacercariae and treated with 1.0 ppm aflatoxin Bl for 12 weeks. Three mice served for untreated-uninfected controls. From 4 weeks after the treatsment and/or in(ection, three mice from each group were sacrificed at 4 week intervals up to the 40th week, and their hepatobiliary tissues were prepared for transmission electron microscopy. The most prominent ultrastructural changes in group I were remarkable enlargement of nuclear size, separation of nucleolus, dispersed chromatin granules in nuclei and increased dense granules along the inner membrane of nuclei. In the cytoplasm there was slight proliferation of mitochondria and endoplasmic reticulum (ER) at earlier stage. At the 12th week separation of fibrillar and granular components of the nucleolus was a characteristic finding. As the time elapsed, epithelial cells showed fiattened-cuboidal form and a tendency of atrophy. Most of the nuclei were elongated and polygonal in shape. In group II the appearance of elaborate interwoven folds of lateral cytoplasm forming a labyrinth of interconnected intercellular space and variety in nuclear shape were the prominent fadings at earlier stage. The cytoplasm showed slight proliferation and dilatation of mitochondria and ER, and a small number of mucin droplets. In the basement membrane scanty fibrous cells were seen. With time, variety in nuclear shape, marked proliferation and dilatation of rough ER and some collagen fibrils were demonstrated. Other features of intracellular organelles and mucin droplets persisted. In group III cuboidal epithelial cells showed their remarkably enlarged and irregular nuclei, increased chromatin granules in the nuclei, separated nucleoli, proliferated and dilated rough ER. With time, sequestered mitochondria showed blob-like evaginations which lacked cristae and dense matrix, and were limited by a single membrane. Since the 20th week, microvilli were relatively scanty and poorly developed. Organelles and inclusions in the cytoplasm of metaplastic cells were poor. Nuclei were variable in shape. The nlost prominent changes at later stage were separation of nuclei from the cytoplasm, and appearance of numerous and irregularly angled electron dense granules in the nuclei.

• Clinical and Experimental Reproductive Medicine
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• v.32 no.3
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• pp.269-277
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• 2005

Objectives: Previously, we sought to compile a list of genes expressed during early folliculogenesis by using cDNA microarray to investigate follicular gene expression and changes during primordialprimary follicle transition and development of secondary follicles (Yoon et al., 2005). Among those genes, a group of genes related to the cell size growth was characterized during the ovarian development in the present study. Methods: We determined ovarian expression pattern of six genes related to the cell size growth (cyr61, emp1, fhl1, socs2, wig1 and wisp1) and extended into CCN family (${\underline{c}}onnective$ tissue growth factor/${\underline{c}}ysteine$-rich 61/${\underline{n}}ephroblastoma$-overexpressed), ctgf, nov, wisp2, wisp3, including cyr61 and wisp1 genes. Expression of mRNA and protein according to the ovarian developmental stage was evaluated by in situ hybridization, and/or semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), and immunohistochemistry, respectively. Results: Among 6 genes related to the cell size growth, cyr61 and wisp1 mRNA was detected only in oocytes in the postnatal day5 mouse ovaries. cyr61 mRNA expression was limited to the nucleolus of oocytes, while wisp1 was expressed in the cytoplasm and nucleolus of oocytes, except nucleus. cyr61 mRNA expression, however, was found in granulosa cells from secondary follicles. The rest 4 genes in the cell size growth group were detected in oocytes, granulosa and theca cells. Cyr61 and Wisp1 proteins were expressed in the oocyte cytoplasm from primordial follicle stage. Especially, Cyr61 protein was detected in pre-granulosa cells, Wisp1 protein was not. By using RT-PCR, we evaluated and decided that Cyr61 protein is produced by their own mRNA in pre-granulosa cells that was not detected by in situ hybridization. cyr61 and wisp1 genes are happen to be the CCN family members. The other members of CCN family were also studied, but their expression was detected in oocytes, granulose and theca cells. Conclusions: We firstly characterized the ovarian expression of genes related to the cell size growth and CCN family according to the early folliculogenesis. Cyr61 protein expression in the pre-granulosa cells is profound in meaning. Further functional analysis for cyr61 in early folliculogenesis is under investigation.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.7
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• pp.956-965
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• 2019

Objective: The main goal of this study was to provide a morphological indicator that could be used to select high-quality oocytes of appropriate meiotic and developmental capabilities in pig. The higher quality of immature oocytes, the higher success rates of in vitro maturation (IVM) and in vitro fertilization (IVF). Thus, prior to the IVM culture, it is important to characterize oocytes morphologically and biochemically in order to assess their quality. Two of the largest indicators of oocyte quality are the presence of cumulus cells and status of chromatin. To investigate the effects of porcine oocyte chromatin configurations on the developmental capacity of blastocysts, we assessed oocyte chromatin status according to follicle size and measured the developmental potency of blastocysts. Methods: To sort by follicle size, we divided the oocytes into three groups (less than 1 mm, 1 to 3 mm, and more than 3 mm in diameter). To assess chromatin configuration, the oocytes were assessed for their stages (surrounded nucleolus [SN] germinal vesicle [GV], non-surrounded nucleolus [NSN] GV, GV breakdown, metaphase I [MI], pro-metaphase II [proMII], and metaphase II [MII]) at different maturation times (22, 44, and 66 h). To assess the development rate, oocytes of each follicle size were subjected to parthenogenetic activation for further development. Finally, GV oocytes were grouped by their chromatin configuration (SN, SN/NSN, and NSN) and their global transcriptional levels were measured. Results: SN GV oocytes were more suitable for IVF than NSN GV oocytes. Moreover, oocytes collected from the larger follicles had a greater distribution of SN GV oocytes and a higher developmental capacity during IVM, reaching MII more quickly and developing more often to blastocysts. Conclusion: Porcine oocytes with high-level meiotic and developmental capacity were identified by analyzing the relationship between follicle size and chromatin configuration. The porcine oocytes from large follicles had a significantly higher SN status in which the transcription level was low and could be better in the degree of meiotic progression and developmental capacity.