• Korean Journal of Fisheries and Aquatic Sciences
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• v.38 no.2
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• pp.94-99
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• 2005

The gametogenesis of the surf clam, Tresus keenae, were investigated by SEM and TEM. Both the testis and the ovary had follicle tubes surrounded by inter-tubal tissue composed of adipogranular cells that provided storage function. In the vitellogenic oocyte, lipid droplets and lipid yolk granules were found in the vacuoles formed by the Golgi apparatus. Proteid yolk granules were formed by the endoplasmic reticulum and cortical granules in the cytoplasm during vitellogenesis. The mature sperm was primitive and resembled a jar with a cover. The sperm heads were approximately $2.00-2.30 {\cal}um$. The acrosomal rod projected in front of the acrosome. In addition, four large mitochondria were in the midpiece.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.112-112
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• 2002

Under normal conditions, women produce a single dominant follicle that participates in a single ovuation each menstrual cycle. But Polycystic ovary syndrome(PCOS) conditions, folliculogenesis does not proceed normally. This condition leads to the accumlation of large numbers of small graffian follicles in which the theca interstitial cells (TIC) produce abnormally large amounts of androgen. PCOS is probably the most common endocrine disorder, affecting women of reprodutive age with 5-10% prevalence estimate. Chronic anovulation, hyperandrogenism, hirsutism, obesity, infertility and polycystic ovaries are clinical hallmarks of women with PCOS. Its etiology remains unknown. To investigate the gene expression pattern of ovary in PCO-induced rat, we used cDNA expression analysis. Total RNA was extracted from the ovary of PCO-induced rat and reverse-transcribed in the presence of［$\alpha$$^{32}$P］-dATP Which were hybridized to Atlas$^{TM}$ Rat Toxicology 1.2 array (Clontech) representing approximately 1176 rat genes. We compared gene expression between ovary of pco-induced immature female rats and control. Differential gene expression profiles were revealed (LIFR-alpha, ADRA1A, Heat shock 90-kDa protein A, PDGFRA). Reverse transcription-polymerase chain reaction(RT-PCR) was used to validate the relative expression pattern obtained by the cDNA array. The precise relationship between the altered expression of genes and PCO is a matter of further investigation. This study was supported by Korea Science and Engineering Foundation(KOSEF)

• Korean Journal of Animal Reproduction
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• v.21 no.3
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• pp.265-274
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• 1997

The purpose of this study was attempted to investigate the a, pp.arences of healthy or artretic follicles in ovaries following repeats of pregnant mare serum gonadotropin(PMSG) treatments for superovulation in nulliparous rats. Thirty two rats(Sprague-Dawely, about 200-250 gm) were randomized into 4 groups. Control group rats were sacrified at estrus phase confirmed by vaginal smear. PMSG-treated group 1 rats, PMSG-treated group 2 rats and PMSG-treated group 3 rats were sacrified at 48 hrs after injection once with PMSG 25 IU, after 2 repeated injection by a week interval, and 3 repeated injection, respectively. The ovaires of rats were removed and then sections by paraffin embedding were stained with H-E or immunohistochemical staining using proliferating cell nuclear antigen monoclonal antibody (PCNA m Ab) and apoptotic kit. The criteria of follicle classification was based as small follicles with preantral follicles with 2~4 layers of granulosa cells surrounding the oocyte, as secondary follicles with more than 5 layers of granulosa cells and early signs of antral cavity or with small clefts on either side of the oocytes, and as tirtiary follicles with a single medium sized antral cavity or large well-formed antral cavity, respectively. The proportions of atretic follicles from small and middle follicles in immunohistochemical staining using PCNA m Ab were 17.9% and 21.3% in control group, 15.5% and 23.5% in PMSG-treated group 1, 24.3% and 26.7% in PMSG-treated group 2, 18.1% and 30.2% in PMSG-treated group 3, respectively. Groups with atretic follicles of higher proportion were ordered as PMSG-treated group 3, PMSG-treated group 2, PMSG-treated group 1 and control group. The proportions of positive cells in small, middle and large follicles were 31.1%, 33.5% and 28.5% respectively. The follicles with positive cells of higher proportion were ordered middle, small and large follicles. In immunohischemical staining using apoptotic kits, small follicles in all 4 groups did not contain positive cells, and proportions of atretic follicles from middle and large follicles were 24.9, 30.7, 33.8 and 40.1% in control, PMSG-treated gruop 1, PMSG-treated group 2 and PMSG-treated group 3, respectively. These results suggested that repeats of PMSG treatment increased proportion of atretic follicles in ovaries, and middle follicles are more quickly developing than small or large follicles.

• Clinical and Experimental Reproductive Medicine
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• v.47 no.3
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• pp.194-206
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• 2020

Objective: The aim of this study was to investigate microRNAs (miRNAs) related to follicle-stimulating hormone (FSH) responsiveness using miRNA microarrays and to identify their target genes to determine the molecular regulatory pathways involved in FSH signaling in KGN cells. Methods: To change the cellular responsiveness to FSH, KGN cells were treated with FSH receptor (FSHR)-specific small interfering RNA (siRNA) followed by FSH. miRNA expression profiles were determined through miRNA microarray analysis. Potential target genes of selected miRNAs were predicted using bioinformatics tools, and their regulatory function was confirmed in KGN cells. Results: We found that six miRNAs (miR-1261, miR-130a-3p, miR-329-3p, miR-185-5p, miR-144-5p and miR-4463) were differentially expressed after FSHR siRNA treatment in KGN cells. Through a bioinformatics analysis, we showed that these miRNAs were predicted to regulate a large number of genes, which we narrowed down to cytochrome P450 family 19 subfamily A member 1 (CYP19A1) and estrogen receptor alpha (ESR1) as the main targets for miR-4463. Functional analysis revealed that miR-4463 is a regulatory factor for aromatase expression and function in KGN cells. Conclusion: In this study, we identified differentially expressed miRNAs related to FSH responsiveness. In particular, upregulation of miR-4463 expression by FSHR deficiency in human granulosa cells impaired 17β-estradiol synthesis by targeting CYP19A1 and ESR1. Therefore, our data might provide novel candidates for molecular biomarkers for use in research into poor responders.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.1
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• pp.45-56
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• 2002

Objectives: Recently, recombinant FSH (rFSH) has been manufactured using a Chinese hamster ovary cell line transfected with the gene encoding human FSH. Both rFSH and urinary gonadotropin (uFSH) could be used for controlled ovarian hyperstimulation (COH). However, uFSH implies a number of disadvantages, such as batch-to-batch inconsistency, no absolute source control, dependence on large amounts of urine, low specific activity, and low purity. The purpose of this study was to evaluate the efficacy of rFSH in human IVF-ET program. Materials and Methods: A total of 508 infertile women was enrolled in this study. They are classified into rFSH group (n=177) or uFSH group (n=331), and all of them were matched by age and cause of infertility in same period. The $Puregon^{(R)}$ (Organon, Holland) was used as rFSH, and the Metrodin-$HP^{(R)}$ (Serono, Switzeland) and $Humegon^{(R)}$ (Organon, Holland) was used as uFSH. We subdivided the patients into three age groups. The outcomes of IVF-ET program were analyzed using the statistical package for social sciences (SPSS). Results: There was no significant differences in the level of estradiol on hCG injection day, the numbers of retrieved oocytes, matured oocytes, fertilized oocytes, transferred embryos, frozen embryos between the two groups. The total dose (IU) of gonadotropin for COH was significantly lower in the rFSH group compared to uFSH group ($1339{\pm}5491.1$ vs $2527.8{\pm}1075.2$ IU, p<0.001). Clinical pregnancy rate per embryo transfer in the rFSH group showed increasing tendency, compared to the uFSH group, but there was no statistical significance (35.2% vs 29.3%). Our results demonstrated that the relative efficiency of rFSH compared with uFSH is higher in older patients. Conclusions: The ovarian stimulatory effect and clinical outcome of recombinant FSH was similar to that of the urinary gonadotropin. The IVF-ET cycles with significantly lower dose of gonadotropin in rFSH group showed comparable results. Therefore, we suggest that recombinant FSH is more potent and effective than urinary gonadotropin.

• Reproductive and Developmental Biology
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• v.33 no.3
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• pp.153-156
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• 2009

The purpose of this study was to assess for re-breeding concentrate period in postpartum in milking cows. The 48 cows aged $3.5{\sim}5.5$ years and of $400{\sim}600\;kg$ body weight were examined every 3rd day from 15 to 36 day postpartum. Blood samples for progesterone and estradiol $17{\beta}$ hormone analyses were withdrawn from the coccygeal vein every third day until the end of the experiment. The ovarian follicular numbers were verified and measured using a multi frequency probe. The least squares means are presented for each day by GLM of SAS. The results showed that ovary lengths (right ovary; $1.64{\pm}0.62\;cm$, left ovary; $1.44{\pm}0.46\;cm$) were similar in right and left ovary activity level during estrous cycle of postpartum cows. We were judged completed uterus on day at $2.31{\pm}0.17\;cm$ level of cervix diameter. And we were monitoring started at $6.44{\pm}2.03\;cm$ from day 15 after postpartum. The results showed that mean plasma concentration of progesterone (3.28 ng/ml) in large follicle gradually increased days 30 in postpartum. And, monitoring of estradiol 176 (22.18 pg/ml) hormone during postpartum period would be useful to predict the ovarian and uterus activity for re-breeding in postpartum milking cows. From these results, we conclude that cervix diameter (mean: 2.31 cm) was very important for reproductive organ recovery standard level of postpartum milking cows, hormone secretion level ($P_4$: 3.28 ng/ml, $E_2$: 22.18 pg/ml) and body condition score ($2.5{\sim}2.75$) level about 30 days in postpartum period.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.30-30
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• 2001

The culture of preantral follicles has important biotechnological implications through its potential to produce the large quantity of oocytes for embryo production, transgenesis research, conservation of rare breed, and a potential source of ovarian genetic material. The present study was conducted to establish the optimal conditions of in vitro culture for intact bovine preantral follicles; and to examine the developmental ability of oocytes derived from the in vitro-grown preantral follicles; and to investigate the effects of various concentrations of FSH and LH on these processes. Bovine preantral follicles (150 $\pm$ 1.2${\mu}{\textrm}{m}$), surrounded by theca cell, were isolated enzymetically and mechanically from ovarian cortical slides in Leibovitz L-15 medium containing 1 mg/$m\ell$ collagens and 0.2 mg/$m\ell$ DNase I and cultured for 25 days in the presence of different concentrations of bovine FSH and LH in $\alpha$MEM medium with insulin, transferrin, and selenite. The survival was tested by frypan Blue and Hematoxylin. The survival and growth rates of follicles were higher in FSH treatment groups than these in control (P<0.001), but there were no significant differences between the LH treatment groups and the control. In 25 days, the survival and growth rates of follicles in FSH and LH treatment group (50%, 300$\pm$1.0${\mu}{\textrm}{m}$) were higher than in FSH treatment group (40%, 244$\pm$0.5${\mu}{\textrm}{m}$) and the control group (25%, 160$\pm$ 1.0${\mu}{\textrm}{m}$). Fifty-five percent of healthy antral follicles were obtained, and 60% of the oocytes complete meiotic maturation to the metaphase II stage. Twenty-two percent of the mature oocytes underwent cleavage, and 9% developed to the blastocyst stage. In this study, in vitro-grown oocytes (111 $\pm$ $1.5mutextrm{m}$), under our culture conditions, were not equivalent in size to the in vivo-grown oocytes (130$\pm$1.3${\mu}{\textrm}{m}$). Therefore, these results suggest that bovine preantral follicles with intact theca cell can grow to the antral stage in 25days, and that oocytes from those follicles can acquire the meiotic competence and normally undergo fertilization and development to the blastocyst stage. However, the developmental capacity of in vitro-grown oocytes is presumably not comparable to those of the in vivo counterparts.

• Applied Microscopy
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• v.10 no.1_2
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• pp.27-32
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• 1980

A case of cutaneous leishmaniasis developed in a 48 year old Korean male who returned from middle east was studied by light and electron microscopic observations. Light microscopically, the lesion consisted of heavy chronic ill-defined granulomatous inflammation involving entire thickness of the dermis, composed of mainly histiocytic and small mononuclear cell infiltrations without evidence of necrosis or giant cell formation. Giemsa staining revealed numerous intracellular micro-organisms within histiocytes, showing dark stained central dot surrounded by light stained cytoplasm. Electron microscopically, the organisms were observed mostly ovoid in shape and frequently binary mitotic features within the host cells. follicle consisted of double unit membranes and microtubules, which are immediately below these membrnae. A long kinetoplast was noted within a very elongated mitochondrion at the center of the organisms and a flagella rose in front of the kineoplast but ended within the cytoplasm. Large numbers of free ribosomes, occasional Golgi complex and SER were also noted, but RER was seldom found. These ultrastructural features corresponded to promastigote stage of Leishmania tropica. In principle, leishmaniasis is a tropical disease and can not be found in temperate zone. However, travel to mideast by many Koreans may contract this disease while they are in endemic regions.

• Journal of Veterinary Clinics
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• v.18 no.1
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• pp.55-60
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• 2001

Immunohistochemical and Electron Microscopical Studies on the Initial Skin Lesions Induced Experimentally by Very Virulent Strain of Marek\`s Disease Virus in Chickens Marek\`s disease virus (MDV), which is an avian herpesvirus, causes malignant CD3+CD4+CD8-T cell lymphomas at many sites including visceral organs, muscles, peripheral nerves and skin. In the early skin lesions induced by MDV, corelationship between the translational activity of MDV early gene, pp38 and demonstration of MDV particles in the lymphoid cells are not well studied. Therefore, skin biopsies taken at weekly intervals for 2 weeks from the same specific-pathogen free chicknes inoculated with Md/5 MDV were examined immunohistochemically and electron microscopically. In the skin biopsies sampled at 1 week and 2 weeks post inoculation (PI), feather follicle epithelium (FFE) exhibited usually strong positive reaction for pp38, whereas only few lymphoblasts, which were infiltrated around FFE revealed positive reaction. Electron microscopically, small lymphocytes were detectable in the dermis and subcutaneous skin tissues sampled at 1 week PI. The number of small lymphocytes was increased and pleomorphic lymphoblasts, which were medium to large in size were scattered among the small lymphocytes at 2 weeks PI. Some of lymphoblasts revealed degenerative and necrotic changes. FFE contained a lot of MDV particles in the nucleus including mature and immature ones. Infrequently, immature virus particles were observed not only in the degenerative and necrotic lymphoblasts, but also rarely in the health lymphoblasts. From the present results, spontaneous MDV activation including translational activity of MDV pp38 gene and formation of MDV particles was occurred in the lymphoblasts of early MD skin lesions.

• Applied Microscopy
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• v.32 no.4
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• pp.303-310
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• 2002

Spermatogenesis and sperm ultrastructure are investigated by means of light and transmission electron microscopy in the equilateral venus, Gomphina veneriformis which is dominant bivalve in the east coast of Korea. In the active spermatogenic season, testis consists of numerous spermatogenic follicles which is contains germ cells in the different developmental stage. The spermatogonia attached to spermatogenic follicle wall and has a large nucleus with electron-dense nucleolus. The spermatocytes are characterized by appearance of synaptonemal complex and well-developed Golgi complex. Nucleus of spermatid consists of numerous heterogeneous granules with high electron density. Karyoplasmic condensation, acrosome and flagellum formations are observed during spermiogenesis. Testicular matured sperms of sperm bundle consists of head, midpiece and tail. The head is about $8.5{\mu}m$ long and comprises a long nucleus and a bullet-like acrosome ($8.5{\mu}m$ in length). Acrosomal rod of microfilaments is observed in the lumen between nucleus and acrosome. The midpiece has four mitochondria. And tail has the typical '9+2' microtubule system.