• Journal of Photoscience
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• v.9 no.2
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• pp.272-274
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• 2002

The anterior brain vesicle of ascidian larvae contains two distinct pigment cells. Ultrastructure of these pigment cells has been shown that the anterior pigment cell is an otolith for perception of gravity and the posterior pigment cell is an ocellus for light reception. The larva has remarkably simple central nervous system (CNS) composed of about 330 cells. We focused to study neural networks of visual systems. In the present paper, we report the whole structure of the photoreceptors of the ascidian larva visualized by an antibody against arrestin. Visual arrestin is the key protein for the termination of phototransduction and one of the abundant proteins in photoreceptors. Recently, we cloned an arrestin homologue gene, Ci-arr and the expression of Ci-arr was found to be restricted to the photoreceptors in the ocellus. To study the whole structure of the photoreceptors in the larva, we prepared an antibody against Ci-Arr. It is found that anti Ci-Arr antibody specifically stains the photoreceptors, including the cell bodies, the axons, and the nerve terminals. The photoreceptor cell bodies lies in row outside the pigment cup which penetrate the pigment cell and is continuous with the outer segments of the photoreceptor cell, inside the concavity of the pigments. The axons form bundle into a single tract. The tract extends toward the midline, where the nerve terminals diverge and seem to form synapses

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.8
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• pp.1190-1195
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• 2007

This study was conducted to improve efficiency of a follicle culture system without reducing developmental competence of intrafollicular oocytes. Preantral follicles (100 to $125{\mu}m$ in diameter) of F1 hybrid (B6CBAF1) mice were cultured singly for 216 h in modified ${\alpha}$-MEM-glutamax medium, to which 2.5 IU/ml hCG and epidermal growth factor was added 16 h prior to the end of culture. Medium change was either performed three times (54 h interval), twice (72 h interval), once (108 h interval), or not at all (216 h interval). Maturation (progression to the metaphase II stage) of intrafollicular oocytes was detected from 4 days after culture in the three-times change treatment, while all treatments yielded mature oocytes from day 5 of culture. Compared with the three-times change, decreasing the change frequency to once did not reduce the capacity to begin maturation (germinal vesicle breakdown of 82 to 86%), to mature (78 to 79%) and to develop into blastocysts after parthenogenetic activation (29 to 32%). Morphological parameters were similar among these treatments. Except for the no medium change treatment, similar colony-forming activity of inner cell mass cells after culturing of blastocysts in leukemia inhibitory factor-containing medium was detected, while the morphology of the colony-forming cells deteriorated in the change-once treatment compared with the change twice or three-times. In conclusion, the efficiency of the preantral follicle culture system could be improved by reducing frequency of medium change up to a 72 h interval (three times in total 216 h culture) without decreasing developmental competence of oocytes.

• Clinical and Experimental Reproductive Medicine
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• v.32 no.1
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• pp.65-70
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• 2005

본 연구는 이식 후 남은 잉여의 포배기 배아를 두 번의 냉동과 융해 과정을 반복적으로 실시한 후 이식한 결과에 관한 보고이다. 사람 포배기 배아의 동결보존에서 높은 생존율과 성공적인 임신율이 보고되고 있으나 미성숙 난자로부터 발달한 포배기 배아에 두 번의 초급속 냉동 방법을 실시한 후 이식한 보고는 되어 있지 않다. 이에 본 연구에서는 다낭성 난소 증후군 환자에게서 얻은 미성숙 난자로부터 발달한 포배기 배아를 artificial shrinkage 후 초급속 냉동함으로써 생존율을 높이는 방법을 이용하여 재냉동 이식하였을 때 임신에 성공한 증례를 보고하고자 한다. 29세의 환자로부터 채취한 55개의 미성숙 난자들(germinal vesicle stage oocytes)을 체외배양 하여 성숙한 37개의 난자들로부터 30개의 수정란을 얻을 수 있었다. 12개의 배아가 포배기 배아까지 발달하였으며 이 중 3개의 양질의 포배기 배아를 선별하여 이식하였고, 이식을 한 후에 남은 9개의 포배기 배아들은 artificial shrinkage의 과정을 마친 후에 초급속 냉동 방법을 이용하여 동결보존 하였다. 그 중, 4개의 포배기 배아들을 융해한 후 이식을 하지 않고 다시 재냉동을 하여 보관하였고 이 후 재냉동 되었던 4개의 포배기 배아들을 다시 융해 하여 이식을 한 결과 임신이 되어 건강한 남아를 분만하였다. 이로써 미성숙 난자로부터 얻은 포배기 배아가 두 번의 냉동과 융해의 과정을 통해 크게 손상을 입지 않고 생존할 수 있다는 것을 알 수 있었다. 그러므로 융해이식 후 남은 잉여의 포배기 배아를 다시 냉동 보관하여 다음 주기에 이용함으로써 축적된 임신율을 증가시킬 수 있을 것으로 사료된다.

• Korean Journal of Environment and Ecology
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• v.17 no.1
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• pp.18-25
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• 2003

In order to know the effects of PCB(Arochlor 1248) on the oocyte maturation and proges-terone(P$_4$) production by FPH(Frog pituitary homogenate: 0.01 p.e/$m\ell$) of frog in vitro, the oocytes were cultured for 20 hours in presence of the PCB at various concentrations and exam-ined their maturation(germinal vesicle breakdown: GVBD) rates and P$_4$ levels secreted by the oocyte in the culture medium. The results show that PCB concentration of 10 ppb suppressed the maturation of the oocytes and secretion of P$_4$. To examine the reversibility of the inhibitory effects, the oocytes were exposed to the PCB only for 3 hours, and then transferred to plain medium and cultured further for 17 hours. The oocytes were recovered from the toxic effect of the PCB when they were exposed to 2.5 ppb, but not to 5 ppb of the PCB. These results indi-cate that PCB suppress the maturation of oocytes and secretion of p, at low concentration, sug-gesting that the frog oocyte culture system can be used as a useful tool to evaluate the toxicity of the pollutants in the environment.

• The Korean Journal of Cytopathology
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• v.1 no.1
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• pp.74-84
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• 1990

Herpes simplex virus type 1 and 2(HSV-1, HSV-2) are the ubiquitous human pathogens responsible for a variety of afflictions. HSV-2 is one of the viruses that were suspected of promoting carcinogenesis in the uterine cervix. Certainly, there is a need for the more sensitive and accurate laboratory techniques for HSV detection. We examined total 80 cases of smears including 17 Tzanck smears of skin and 63 cases of Papanicolaou smears from total 77 patients with clinical impression of herpetic infections, from September, 1985 through August, 1989. Immunohistochemical typings for HSV-1 and HSV-2 were performed together with routine cytologic findings and compared. The results are as follows : 1) Patients were 9 males and 33 females, and age distribution was between 5 and 71 years. 2) Subjective symptoms such as ulceration, vesicle, vaginal discharge, pruritus, and pain were complained in 36 patients and 38 cases were genital herpes. Recurrence was noted in 11 cases. 3) Positive results were obtained in 42 among 80 cases. 4) Both routine cytology and immunohistochemical staining were positive in 13 cases and in 24 cases only immunohistochemical staining were positive. 5 cases were positive only in routine cytologic smears. 5) The cases that immunocytochemical stain had been performed were 37 cases, which were all positive in type 2. Among the above 37 cases, type 1 also were positive in 5 cases. The results show that the immunoperoxidase technique is one of the rapid and reliable method to confirm the herpetic infection when suspected and that it is particularly useful when the Papanicolaou smear findings are equivocal.

• Toxicological Research
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• v.16 no.2
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• pp.163-172
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• 2000

Recently, there is a worldwide concern that a great number of man-made chemicals have a hormone-like action both in humans and in animals. DECD is developing screening programs using validated test systems to determine whether certain substances may have an effect in humans. In the present study. the establishment oj repeated-dose toxicity test method was tried. Flutamide. an anti-androgenic agent. was administered by gavage to Sprague-Dawley rats for 28 days at dose levels of 0. 0.5. 3 and 18 mg/kg body weight (10-15 rats/sex/group) to examine the effects on general findings. especially reproductive and endocrine parameters. Clinical signs. body weights, food consumption, and sexual cycle were checked and measured. For the gross and microscopic examinations. 10 rats/sex/group were sacrificed at the end of dosing period and the remaining animals of control and high dose groups (5 each) were sacrificed after 14 days recovery. Examinations for hematology and clinical chemistry were carried out at necropsy. There were no treatment-related changes in clinical signs. body weights, food consumption. gross necropsy. hematology and clinical chemistry at all doses of both sexes. The period and regularity of sexual cycle were not adversely affected at all doses by the test agent. At 18 mg/kg. both decreased weights of prostate, seminal vesicle and epididymis in males and increased weights of spleen and thymus in females were observed. In addition, decreased number of spermatids and sperms. increased serum testosterone concentration and increased incidence (100%) of interstitial cell hyperplasia were seen in males. At 18 mg/kg of the recovery group. decreased prostate weight. reduced sperm count and increased incidence (20%) of interstitial cell hyperplasia in males and increased thymus weight in females were observed. At 3 mg/kg. reduced sperm count was found. There were no adverse effects on parameters examined at 0.5 mg/kg of both sexes. The results suggested that the potential target organs of flutamide may be accessory sexual glands including testes for males and spleen and thymus for females. Taken together. this test method was found to be a useful screening test system for endocrine disrupting chemicals.

• The Korean Journal of Physiology
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• v.30 no.2
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• pp.279-288
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• 1996

Chronic exposure to cadmium impairs various renal tubular functions, including organic acid (anion) secretion. To investigate the mechanism of cadmium-induced alterations in the organic anion transport system, kinetics of p-aminohippurate (PAH) uptake was studied in renal cortical basolateral membrane vesicles (BLMV) isolated from cadmium-intoxicated rats (adult male Sprague-Dawley). Cadmium intoxication was induced by subcutaneous injections of $CdCl_{2}$ (2 mg Cd/kg per day) for 3 weeks. The renal plasma membrane vesicles were prepared by Percoll gradient centrifugation. The vesicular uptake of $^{14}C$-PAH was determined by rapid filtration technique using Millipore filter. Cadmium intoxication resulted in a marked attenuation of $Na^{+}$-dependent, ${\alpha}$-ketoglutarate (${\alpha}$KG)-driven PAH uptake with no changes in $Na^{+}$ and ${\alpha}$KG-independent transport component. Kinetic analysis indicated that Vmax, but not Km, of the $Na^{+}$-dependent, ${\alpha}$KG-driven component was reduced. A similar reduction of $Na^{+}$-dependent, ${\alpha}$KG-driven PAH uptake was observed in normal membrane vesicles directly exposed to inorganic cadmium in vitro, and this was accompanied by an inhibition of both $Na^{+}$-dependent ${\alpha}$KG uptake and ${\alpha}$KG-PAH exchange activity. These results indicate that during chronic exposure to cadmium, free cadmium ions liberated in the proximal tubular cytoplasm directly interact with the basolateral membrane and impair the active transport capacity for organic anions, most likely due to an inhibition of both $Na^{+}$-dicarboxylate cotransporter and dicarboxylate-organic anion antiporter activities.

• Reproductive and Developmental Biology
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• v.36 no.4
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• pp.237-241
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• 2012

Embryonic genome activation (EGA) is the first major transition that occurs after fertilization, and entails a dramatic reprogramming of gene expression that is essential for continued development. Although it has been suggested that EGA in porcine embryos starts at the four-cell stage, recent evidence indicates that EGA may commence even earlier; however, the molecular details of EGA remain incompletely understood. The RNA polymerase II of eukaryotes transcribes mRNAs and most small nuclear RNAs. The largest subunit of RNA polymerase II can become phosphorylated in the C-terminal domain. The unphosphorylated form of the RNA polymerase II largest subunit C-terminal domain (IIa) plays a role in initiation of transcription, and the phosphorylated form (IIo) is required for transcriptional elongation and mRNA splicing. In the present study, we explored the nuclear translocation, nuclear localization, and phosphorylation dynamics of the RNA polymerase II C-terminal domain in immature pig oocytes, mature oocytes, two-, four-, and eight-cell embryos, and the morula and blastocyst. To this end, we used antibodies specific for the IIa and IIo forms of RNA polymerase II to stain the proteins. Unphosphorylated RNA polymerase II stained strongly in the nuclei of germinal vesicle oocytes, whereas the phosphorylated form of the enzyme was confined to the chromatin of prophase I oocytes. After fertilization, both unphosphorylated and phosphorylated RNA polymerase II began to accumulate in the nuclei of early stage one-cell embryos, and this pattern was maintained through to the blastocyst stage. The results suggest that both porcine oocytes and early embryos are transcriptionally competent, and that transcription of embryonic genes during the first three cell cycles parallels expression of phosphorylated RNA polymerase II.

• Development and Reproduction
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• v.17 no.4
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• pp.469-475
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• 2013

In mammals, puberty is a process of acquiring reproductive competence, triggering by activation of hypothalamic kisspeptin (KiSS)-gonadotropin releasing hormone (GnRH) neuronal circuit. During peripubertal period, not only the external genitalia but the internal reproductive organs have to be matured in response to the hormonal signals from hypothalamic-pituitary-gonadal (H-P-G) axis. In the present study, we evaluated the maturation of male rat accessory sex organs during the peripubertal period using tissue weight measurement, histological analysis and RT-PCR assay. Male rats were sacrificed at 25, 30, 35, 40, 45, 50, and 70 postnatal days (PND). The rat accessory sex organs exhibited differential growth patterns compared to those of non-reproductive organs. The growth rate of the accessory sex organs were much higher than the those of non-reproductive organs. Also, the growth spurts occurred differentially even among the accessory sex organs; the order of prepubertal organ growth spurts is testis = epididymis > seminal vesicle = prostate. Histological study revealed that the presence of sperms in seminiferous tubules and epididymal ducts at day 50, indicating the puberty onset. The number of duct and the volume of duct in epididymis and prostate were inversely correlated during the experimental period. Our RT-PCR revealed that the levels of hypothalamic GnRH transcript were increased significantly on PND 40, suggesting the activation of hypothalamic GnRH pulse-generator before puberty onset. Studies on the peripubertal male accessory sex organs will provide useful references on the growth regulation mechanism which is differentially regulated during the period in androgen-sensitive organs. The detailed references will render easier development of endocrine disruption assay.

• Development and Reproduction
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• v.17 no.4
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• pp.435-440
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• 2013

Previous studies, including our own, have demonstrated that the intratesticular injection of hypertonic saline (20%) decreased serum testosterone level which was similar to the surgical castration in the rat, showing the state of chemical castration. In the present study, we further verify the efficacy of this less invasive method as an alternative of surgical orchidectomy in the andrological field. Sterilized 20% saline was directly injected into the adult male rats (750 ${\mu}l$ per testis). The tested rats were divided into 3 groups including intact group (intact), orchidectomy group (ORX) and saline injection group (SAL) after bilateral orchidectomy was performed at the same day of injection. All rats were sacrificed at 4 weeks after injection. The reproductive organs (testes, epididymis, seminal vesicles and prostates) were collected and used for DNA and protein pattern analyses. Also, patho-histological studies on the testes were performed. In contrast to the intact group, similar DNA damages of testis and seminal vesicle were appeared in ORX group and SAL group. The DNA degradations seemed to be the results of necrosis rather than apoptosis. In the protein pattern analysis, all the testing tissues exerted similar patterns in the ORX group and the SAL group compared to the those of intact group. Patho-histological studies revealed that severe degenerative changes in testicular seminiferous tubules and massive infiltration of immune cells in SAL group. The present study confirmed that direct injection of hypertonic saline into the testis caused the equivalent biochemical changes in the accessory sex organs as shown in the orchidectomized animals. These results suggest that hypertonic saline injection model could be a useful castration model which can substitute for surgical castration when its safety is secured through further study in the future.