• Journal of Embryo Transfer
• /
• v.31 no.3
• /
• pp.191-197
• /
• 2016

Historically, Korea old cattle had been consisted with various lines of coat color brindle, black and white-brown breeds or more. The two rare lines of black and white coat color are maintained for animal resources and preserved critically. The present study was carried out to evaluate potential usage of cysteamine supplementation during in vitro matration (IVM) and in vitro culture/production of embryo (IVP) by transvaginal ultrasound-guided follicle aspiration (Ovum Pick-Up: OPU) for the establishment of cryo-banking system. Immature slaughterhouse-derived cumulus-oocyte complexes (SL-COCs) were matured in IVM medium supplemented with 0, 0.1, 0.3 or 0.9 mM cysteamine, and then cultured in mSOF-BAS for 8 days after in vitro fertilization. The treatment of 0.1 mM cysteamine on SL-COCs showed higher rate of blastocyst, so OPU-derived COCs from rare breeds were matured in TCM media supplemented with or without 0.1 mM cysteamine, FSH and 5% FBS. The embryos were evaluated their developmental stages on day 8. During IVM, cysteamine treatment significantly increased the embryo production rate of slaughterhouse-derived COCs (19.6% vs. 30.5%). The presence of cysteamine during IVM of OPU-derived COCs from rare Korean cattle breeds (albino white and black line) also increased embryo production rates than those from SL-COCs (27.4% vs. 41.9% and 36.4%). With these results, cysteamine treatment during IVM is one of key factors IVP of blastocysts to establish banking system of endangered rare Koarean cattle with OPU derived transferable blastocysts.

• Journal of Life Science
• /
• v.28 no.7
• /
• pp.811-818
• /
• 2018

This study was performed to investigate the localization and concentration changes of mercury compound in female reproductive organs with time. Methylmercuric chloride was subcutaneously injected weekly into pubescent female mice for 3 weeks. For the concentration changes of mercury with time, the mice were sacrificed at 10, 150, and 300 days post treatment (DPT). Body and organ weights were not significantly different between the control and mercury-treated groups, except for 10 DPT in body weight. Localization of accumulated mercury was identified by the autometallography method. Localization of mercury compounds in the uterus, ovary, and ovum was analyzed with a light microscope. In the uterus, mercury was densely located in the stroma cells and surface epithelium of the perimetrium at 10 DPT. Mercury concentration was decreased at 150 DPT and did not appear at 300 DPT. In the ovary, mercury particles were distributed in the stroma cells of the cortex region, cells of the theca around the follicle, and the corpus luteum at 10 DPT. Mercury was concentrated in the medulla region at 150 DPT and was not distributed at 300 DPT. In the ovum, mercury particles were mainly located in the marginal region at 10 and 150 DPT. Mercury concentration was decreased and evenly distributed at 300 DPT. These results suggest that hormone synthesis, implantation, and developing embryos will be affected by mercury compound in the female mouse.

• The Korean Journal of Malacology
• /
• v.31 no.4
• /
• pp.273-277
• /
• 2015

This study was conducted to provide the reproductive biological information and basic data on the artificial sex control of Haliotis discus hannai. The morphological sex differentiation process of H. discus hannai could be classified into following five phases: 1) formation of gonad outer membrane (FGOM) (${\leq}SL\;10.0{\pm}1.0mm$), 2) primordial germ cells (PGCs) appearance in the connective tissue between intestine and hepatopancreas (PAC), and formation of gonadal cavity (FGC) (SL $15.0{\pm}2.0mm$), 3) PGCs appearance in the epithelial layer of gonadal cavity (PAG) (SL $18.0{\pm}2.0mm$), 4) formation of gametogenic follicle and appearance of early oocytes and spermatogonia (FGOC) (SL $21.0{\pm}2.0mm$), 5) morphological sex differentiation (MSD) (${\geq}SL\;23.0{\pm}2.0mm$). From histological analysis sex differentiation rate in SL 24.1-25.0 mm of H. discus hannai was 90.0% and sex ratio (female : male) was 1:0.8.

• Clinical and Experimental Reproductive Medicine
• /
• v.23 no.1
• /
• pp.103-108
• /
• 1996

Through the previous studies(I,II), it was observed that human follicular fluid(HFF) was more effective than human fetal cord serum(HFCS) on promoting meiotic resumption of oocytes and improving embryonic development of mouse in vitro. On the basis of these results, we have gradually exchanged HFCS with HFF as protein supplement in human ART. This study was performed to investigate the efficiency of HFF on improving the pregnancy rate in ART. Oocytes were retrieved transvaginally from patients treated with pituitary suppression with GnRH-agonist and ovarian stimulation with human menopausal gonadotro-pin(HMG) and pure follicle stimulating hormone(FSH). Aspirated oocytes were rinsed and cultured in TCM-199 containing HFF, and the concentrations of HFF were adjusted to 10, 20, and 30% according to the use for insemination, embryo growth and embryo transfer, respectively. As possible as, we tried to do embryo transfer into fallopian tube to mimic the coincidence of the cell stage with the place of sojourn in vivo, so we performed various ART programs(IVF & ET; in vitro fertilization, ZIFT; zygote intra fallopian-tube transfer, ZIFT & ET) according to the tubal conditions of patients. On the while, intra cytoplasmic sperm injection(ICSI) was used to assist IVF of the patients who had shown poor standard IVF results by immunological or severe male factor. Of the 255 cycles of ART programs using HFF as protein supplement, 118 cycles were turn out to be succeeded in pregnancy(46.2%, per cycle, p<0.05), while 21 pregnancies were achieved in the 69 cycles using HFCS(30.4%). The 255 cycles using HFF were subdivided into cycles with the type of ART programs, and each pregnancy rate of the ART programs were 44.7% (IVF & ET, 76/170 cycles), 53.4%(ZIFT, 31/58 cycles) and 40.7% (ZIFT & ET, 11/27 cycles), respectively. In the 61 ICSI cycles using HFF, 28 cycles succeed in pregnancy(45.9%), while 7 pregnancies were obtained in the 17 ICSI cycles using HFCS. Also the ongoing pregnancy rate in the group using HFF(78.8%, 93/118 cycles) was higher than that in the group using HFCS(61.9%). Therefore, we found that the use of HFF as protein supplement was more suitable and effective than the use of HFCS to improve the pregnancy rate in ART.

• Development and Reproduction
• /
• v.7 no.1
• /
• pp.15-22
• /
• 2003

The Fas antigen (Fas) as a cell-surface receptor protein which mediates apoptosis-inducing signals plays an important role in the immune system. Expression of Fas mRNA is detected not only in lymphoid organs but also in the nonlymphoid organs. In the ovary, most of the follicles is known to undergo atreisa through apoptosis. However, the exact mechanism of atresia was not elucidated yet. Therefore, the purposes of the present study were to investigate the expression of Fas and Fas ligand in mouse ovary and to clarify the relationship between expression of Fas and Fas ligand and atresia of follicle. The result of RT-PCR demonstrated that Fas and Fas ligand mRNA was expressed in ovary, especially granulosa cells and oocytes. The immunohistochemistry showed that the granulosa cells and oocytes in growing follicles were stained for Fas and Fas ligand, but primordial follicles were not. Furthermore, Fas and Fas ligand were intensively stained in the atretic follicles As results of TUNEL staining to detect apoptotic cells in the ovaries, the number of TUNEL-positive (apoptotic) granulosa cells and oocytes increased in the atretic follicles compared to the healthy normal follicles. These results demonstrate that there is the positive relationship between expression of Fas and Fas ligand in granulosa cells and oocyies and apoptosis of them leading to atresia of follicles. It suggests that expression of Fas and Fas ligand could be associated with atresia of follicles in mouse ovary.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.37 no.4
• /
• pp.505-511
• /
• 2010

Odontoma represents 22% of all odontogenic tumors, which is characterized by slow growth pattern. Most of the odontomas usually develop during dental follicle proliferation. The growth of odontoma is limited and lesion is generally asymptomatic. It is frequently diagnosed during assessments for delayed eruption of permanent tooth and is usually founded in the second decade of life. Odontoma is usually diagnosed through radiographic views and is difficult to diagnose at the early developmental stage of odontoma. But an uncalcified developing odontoma can disturb the eruption of the tooth, so it is important to perform periodic radiographic examinations. Treatments are surgical removal and observation of odontoma followed by surgical opening or orthodontic traction of impacted tooth according to the tooth development and the location of impacted tooth. In this case, we found the radiopaque calcified odontoma in the radiographic view meanwhile follow up of the impacted tooth showing idiopathic eruption disturbance. This suggests that a developing odontoma is the cause of eruption disturbace.

• Korean Journal of Environmental Biology
• /
• v.20 no.3
• /
• pp.224-231
• /
• 2002

Photoperiod (length of light per day) is a major factor in regulating reproductive function in golden hamsters. The information of photoperiod is transmitted to the reproductive endocrine system by melatonin. Thus the effects of melatonin aye investigated in male golden hamsters exposed to photoperiods. Paired testicular weights were markedly reduced in the animals housed in short photoperiod $(SP,\le{12\;hours\;day^{-1})$ and injected with melatonin in the evening, but not in long photoperiod $(LP,\le{12.5}\;hours\;day^{-1})$ and injected with melatonin in the morning. The histological examination of regressed testes showed reduction of tubular lumen diameter including the numbers of cells and Leydig cell number. The mean values of both follicle stimulating hormone (FSH) and luteinizing hormone (LH) were also lowered in the sexually inactive animals than in the sexually active animals. Melatonin receptor was identified by reverse-transcription polymerase chain reaction (RT-PCR) and its expression was examined in various tissues to scrutinize the action site of melatonin. It turned out 309 nucleotides and was definitely expressed in hypothalamus and pituitary including spleen, retina, and epididymis. And gonadotropin releasing hormone (GnRH) gene, which is a key element in regulating reproduction, was identified by RT-PCR but the expression of GnRH was not modified by the treatment of melatonin. Taken together, photoperiod via melatonin indirectly affects reproductive endocrine system, possibly through the release of GnRH, not the synthesis of GnRH.

• Development and Reproduction
• /
• v.11 no.1
• /
• pp.13-20
• /
• 2007

In all female mammals, reproductive system is one of the first biological systems to show age-related decline. Female mammals in reproductive aging, though the phenomena is somewhat species-specific, start to show declining fertility and changes of numerous physiological functions. This review will present a current information on the aging of the female reproductive hormonal axis and introduce three useful rodent models for studying this field. Middle age($8{\sim}12$ months old) in female rats and mice is comparable to the stage prior to the entry of menopause in human. In this period pulsatile and surge GnRH secretion from hypothalamus gradually attenuated, then reduced pulsatile and surge LH secretion is followed consequently. This age-related defects in GnRH-LH neuroendocrine axis seem to be highly correlated with the defects in brain signals which modulate the activities of GnRH neuron. Many researchers support the idea which the age-related hypothalamic defects are the main cause of reproductive aging, but some ovarian factors such as inhibin response also could contribute to the induction of reproductive senescence. Some rodent models are quite valuable in studying the reproductive aging. The follitropin receptor knockout(FORKO) mice, both of null and haploinsufficient state, could produce depletion of oocyte/follicle with age. Dioxin/aryl hydrocarbon receptor(AhR) knockout mice also show severe ovarian defects and poor reproductive success early in their life compared to the age-matched normal mice. Further studies on the reproductive aging will be a great help to evaluate the benefits and risks of hormone replacement therapy(HRT) and to improve the safety of HRT.

• Journal of Genetic Medicine
• /
• v.2 no.1
• /
• pp.11-15
• /
• 1998

This paper reports 3 cases with 46,XX sex reversed male. Three 46,XX hypogonadal subjects showed complete sex reversal and had normal phallus and azoospermia. We studied them under clinical, cytogenetic and molecular aspects to find out the origin of the sex reversal. Patients had markedly elevated serum follicle-stimulating hormone (FSH) and lutenizing hormone (LH) and decreased or normal range of serum testosterone. The testicular volumes were small (3-8ml). Testicular biopsy showed Leydig cell hyperplasia and atrophy of seminiferous tubules. We obtained the results of normal 46,XX, and the presence of Y chromosome mosaicism was ruled out through XY dual fluorescent in situ hybridization (FISH). By using polymerase chain reaction (PCR), we amplified short arm (SRY, PABY, ZFY and DYS14), centromere (DYZ3), and heterochromatin (DYZ1) region of the Y chromosome. PCR amplification of DNA from these patients showed the presence of the sex-determining region of the Y chromosome (SRY) but didn't show the centromere and heterochromatin region sequence. The SRY gene was detected in all the three patients. Amplification patterns of the other regions were different in these patients; one had four amplified loci (PABY+, SRY+, ZFY+, DYS14+), another had two loci (SRY+, ZFY+) and the other had two loci (PABY+, SRY+). We have found that each patient's translocation elements had different breakpoints at upstream and downstream of the SRY gene region. We conclude that the testicular development in 46,XX male patients were due to insertion or translocation of SRY gene into X chromosome or autosomes.

• The Korean Journal of Zoology
• /
• v.30 no.1
• /
• pp.1-9
• /
• 1987

Cyclic AMP (cAMP) was known to play a key role in the regulation of cumulus expansion and oocyte maturation of mammalian cumulus-oocyte complexes (COC's) in vivo and in vitro. The present experiments were conducted to know how intracellular level of cAMP in these cells is controlled. Intracellular cAMP level was modulated by culturing mouse CGC's with an adenylate cyclase stimulator, forskolin, a phosphodiesterase inhibitor, 3-isobutyl-1-methyixanthine (IBMX), human chorionic gonadotrophin (HCG), or follicle stimulating hormone (FSH). The rate of cumulus expansion and germinal vesicle break-down (GVBD) was checked after culture and used as a biological end point. Forskolin in the medium began to stimulate the expansion of the complexes at 1 nM and induced maximum expansion (80~90%) at 0 1~10 $\mu$M. The expansion rate was reduced to 60% when forskolin concentration was increased to 100 $\mu$M. Oocyte GVBD occurred normally (75~82%) in the presence of 10 $\mu$M of forskolin, but partial suppression was appeared at 100 pM of the drug (40%). IBMX also stimulated the expansion from the concentration of 0.01 pM and induced full expansion (81~89%) between the concentration of 1-1000 $\mu$M. Meiotic resumption was occurred normally under 10 $\mu$M of IBMX, but suppressed drastically from the concentration of 100 $\mu$M. The minimum exposing time to hormone or drugs required to trigger cumulus expansion was two minutes with HCG, 15~30 minutes with FSH and fors kolin, and two hours with IBMX. The data presented here seemed to imply that intracellular cAMP level in cumulus cells is regulated by both adenylate cyclase and phosphodiesterase and cumulus expansion is induced by a peak of cAMP while meiotic arrest is maintained by continuous presence of cAMP.