• Journal of Veterinary Clinics
• /
• v.31 no.5
• /
• pp.394-402
• /
• 2014

The objective of this study was to investigate the effect of chronic unpredictable stress on the reproductive function and ovarian luteinizing hormone receptor (LHR) expression. 9-week-old C57BL/6 female mice were randomly divided into two groups: control group and stressed group. Mice have been stressed twice a day for 35 days with 12 different stressors which were randomly selected. The results demonstrate that there is significant increase in the anxiety-related behaviors (P < 0.05), decrease body weight gain rate (P < 0.01) and decrease in the average of litter size in stressed mice compared with control group (P < 0.01). Furthermore, the rate of primary, secondary and early antral follicles in stressed mice significantly decreased (P < 0.05), whereas that of atretic follicles significantly increased compared with control mice (P < 0.01). The immunohistochemical analysis revealed that reduced LHR expression in granulosa cells of follicle and luteal cells of corpus luteum in response to chronic unpredictable stress. The western blot analysis revealed significantly decrease in LHR expression in the stressed mice ovaries compared with the control (P < 0.05). These results suggest that ovarian LHR expression affected by chronic unpredictable stress and the modulated ovarian LHR is responsible for ovarian follicular maldevelopment and reproductive dysfunction.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.19 no.6
• /
• pp.563-574
• /
• 1986

The structure of gonads, gametogenesis and reproductive cycle of the jackknife clams, Solen strictus and Solen gordonis were investigated mainly by histological observation. The first species used were monthly sampled at the coastal area of Dadaepo, Pusan, Korea and Naechodo, Kunsan, Korea for one year from February 1982 to January 1983. The second species were monthly sampled at the sand beach of Dadaepo, Pusan, Korea, from February 1982 to January 1983. Sexualities of Solen strictus and Solen gordonis are dioecious, and these species are oviparous. The gonads are irregularly arranged from the subregion of mid-intestinal gland in visceral cavity to reticular connective tissue of foot. The ovary was composed of a number of small ovarian sacs and the testis was composed of several testicular lobuli which from the tubular structure. Early multiplicating oogonium was about $10{\mu}m$ in diamater. Nucleus and nucleolus, at that time, were distinct in appearance. Each of the early growing oocytes made an egg-stalk, connected to the germinal epithelium of the ovarian sac. A great number of undifferentiated mesenchymal tissue and eosinophilic granular cells are abundantly distributed in the ovarian sacs in the early development stages. With the further development of gonad, these tissue and cells gradually disappeared. Then the undifferentiated mesenchymal tissue and eosinophilic granular cells function as nutritive cells in the formation and development of the early stage germ cells. Mature oocytes were free in the lumen of ovarian sacs and gradually become round or oval. Ripe oocyte was about 80 to $90{\mu}m$ in diameter. With the further development of testis, each of the testicular lobuli formed stratified layers composed of spermatogonia, spermatocytes, spermatids and spermatozoa in groups on the germinal epithelium. After spawning, the gonad gradually degenerated, and disorganized completely. Then new differentiated tissues were rearranged next year. The annual reproductive cycle of those species could be classified into five stages; multiplicative, growing, mature, spent, degenerative and resting stage. It seems that the spawning season is closely related to the water temperature, and the spawning of Solen strictus occurs from June to July at above $20^{\circ}C$ in water temperature. The peak spawning season appeared in June at Dadaepo and in July at Kunsan, The spawning of Solen gordonis occurs from May to June with the peak spawning season in June. Percentages of the first maturity in female of Solen strictus ranging from 5.1-6.0 cm and 7.1-8.0 cm in shell length were $50\%$ and $100\%$, respectively.

• Development and Reproduction
• /
• v.6 no.2
• /
• pp.131-139
• /
• 2002

Since GnRH and its receptor genes are expressed in the ovary, it has been suggested that ovarian GnRH might be involved in the regulation of ovarian function and the apoptosis of ovarian cells. However, it was not known well on the expression and function of GnRH and its receptor in the corpus luteum. The present study was undertaken to investigate whether GnRH and its receptor are expressed in luteal cells and GnRH has any effect on the apoptosis of luteal cells. Luteal cells obtained from the pregnant rats were cultured and stained for GnRH and its receptor proteins. Cultured luteal cells showed distinct immunoreactivity against both anti-GnRH and anti-GnRH receptor antibodies. In addition, the presence of GnRH receptor protein in cultured cells was confirmed by Western blot analysis. To investigate the effect of GnRH on the apoptosis of luteal cells, luteal cells were cultured in the presence of 10$^{-6}$ M GnRH-agonist(GnRH-Ag) for 3, 8, and 12h. TUNEL assay showed that the number of cells undergoing apoptosis increased 12h after culture(P<0.05). DNA fragmentation analysis confirmed the results such that the cells treated for 12h showed the greatest increase of fragmentation(p<0.05). Further, Western blot analysis of cytochrome c in the mitochondrial and cytoplasmic fractions of the luteal cells showed that GnRH-Ag treatment increased the content of cytochrome c in cytoplasm. These results demonstrate that the luteal cells express GnRH and its receptor and GnRH-Ag treatment induces apoptosis of the luteal cells via mitochondrial release of cytochrome c. The present study suggest that the releasing of cytochrome c from mitochondria might be involved in the luteal cell apoptosis induced by GnRH-Ag.

• BMB Reports
• /
• v.50 no.8
• /
• pp.417-422
• /
• 2017

Cisplatin is the most effective and widely used chemotherapeutic agent for many types of cancer. Unfortunately, its clinical use is limited by its adverse effects, notably bone marrow suppression leading to abnormal hematopoiesis. We previously revealed that neuropeptide Y (NPY) is responsible for the maintenance of hematopoietic stem cell (HSC) function by protecting the sympathetic nervous system (SNS) fibers survival from chemotherapy-induced bone marrow impairment. Here, we show the NPY-mediated protective effect against bone marrow dysfunction due to cisplatin in an ovarian cancer mouse model. During chemotherapy, NPY mitigates reduction in HSC abundance and destruction of SNS fibers in the bone marrow without blocking the anticancer efficacy of cisplatin, and it results in the restoration of blood cells and amelioration of sensory neuropathy. Therefore, these results suggest that NPY can be used as a potentially effective agent to improve bone marrow dysfunction during cisplatin-based cancer therapy.

• Korean Journal of Animal Reproduction
• /
• v.16 no.4
• /
• pp.387-397
• /
• 1993

To examine local response of adenosie(purine nucleoside) on the developing mammary gland, Elvax 40P implants containing adenosine were surgically implanted into mammary fat pad of the five week old female ICR mice. Inguinal(the 4th) mammary glands of anesthetized mice were exposed andplaced the implants for 12 days. One gland was treated with an adenosine implant, while the contralateral gland received a blank implant as control. For whole-mount preparations, glands were stained with alum carmine, and for histological observation, micro-selected mammary glands were stained with hematoxylin and eosin Y. Implantation with Elvax 40P did not affect on the damage of neighboring mammary tissue. Adenosie 25 or 250$\mu\textrm{g}$ per slow-release implant stimulated local mammary end bud formation of ovariectomized mice such as end bud size and numbers of end bud per gland in a dose dependent manner(P<0.05), and lower concentration of adenosie(2.5 or 25$\mu\textrm{g}$/implant) increased numbers of end bud(P<0.05) and end bud size(P<0.1) of intact mice. Adenosine treatment and intact ovarian function had moderate interation effects on the stimulation of end bud formation at 2.5$\mu\textrm{g}$ adenosine/implant(P<0.1). In histological observation, adenosine implants increased numbers of mammary epithelial type of cells at mammary duct in the presence or absece ofovary. These results indicate that adenosine should be one of regulators in mouse mammary ductal growth.

• Journal of Microbiology and Biotechnology
• /
• v.13 no.4
• /
• pp.517-521
• /
• 2003

Telomerase is a ribonucleoprotein complex whose function is to add telomeric repeats to chromosomal ends. Telomerase consists of two essential components, telomerase RNA template (hTR) and catalytic subunit (hTERT). hTERT is expressed only in cells and tissues positive for telomerase activity, i.e., tumor and fetal cells. In this report, the possibility of utilization of the hTERT promoter in targeted cancer gene therapy was tested. The hTERT promoter was cloned in the replacement of the CMV promoter, and the HSV-TK gene was subcloned to be controlled by the hTERT gene promoter in the adenovirus shuttle plasmid. Then, the recombinant adenovirus Ad-hT-TK was constructed and was infected into normal and human gynecological cancer cell lines. The selective tumor specific cell death by Ad-hT-TK was identified through these experiments, showing that Ad-hT-TK could be used for targeted cancer gene therapy.

• Korean Journal of Animal Reproduction
• /
• v.20 no.2
• /
• pp.215-221
• /
• 1996

This study was carried out to develop an immunoassay for the diagnosis of the pregnancy and ovarian function of domestic animals. Using 2E92C10 monoclonal antibody(McAb) generated against estrone-3-glucuronide(E1-3-G) and appeared a high cross-reactivity with estrone-3-sulfate(E1-3-S), chemiluminesence immunoassay (CIA) to detect E1-3-S was developed. 2E92C10 McAb cross-reacted with E1-3-S (30%) was purified from ascites fluid using protein G sepharose gel column. The purity of purified antibody fraction was monitored by SDS-PAGE and was better compared to that of crude ascite fluid. The soild and liquid phase CIA for E1-3-S were established utilizing 2E92C10 antibody and E1-3-G-ABEI conjugate used as a tracer. As the results, the titer of 2E92C10 antibody was 5g/ml in soild phase and 1:2000 in liquid phase. The sensitivity on soild and solid phase CIA were about 200 pg/ml. These results indicate that CIA for measurement of E1-3-S was successfully developed by using ant-E1-3-G McAb cross-reacted with E1-3-S and could be usefully used to research this area.

• The Korean Journal of Malacology
• /
• v.4 no.1
• /
• pp.30-41
• /
• 1988

The gonadal development, the annual reproductive cycle and the first sexual maturity of surf clam, Mactra veneriformis Reeve were studied histologically. Speciemens were monthly collected at the intertidal zone of Naechodo, Chollabuk-do, Korea, for one year from March 1986 to February 1987. Sexuality of the clam is dioecious. The gonads were located between the subregion of mid-intestinal gland in the visceral cavity and the reticular connetive tissues of the foot, The ovary is composed of a number of ovarian sacs, and the testis comprise several testiculat lobules. The undifferentiated mesenchymal tissues and eosinophilic granular cells function as nutritive cells in the early stage. The ripe eggs were about 50-60${\mu}{\textrm}{m}$ in diameter, and they were wurroundedby the gelatinous membranes. The spawing period was from early June to September the main spawning occurred beetween July and August when the water temperature reached above 24$^{\circ}C$. The annual reproductive cycle of this species could be classified into five successive stages: multiplicative(January to March), growing(March to May), mature(April to August), spent(June to September), degenerative and resting(September to February). The monthly changes of fatness coefficient closely correlated with the annual reproductive cycle. Percentages of the first sexual maturity of female and male clams were over 50% among those individals ranging from 2.1 To 2.5cm, and 100% in those over 2.6cm in shell length.

• The Journal of Korean Obstetrics and Gynecology
• /
• v.25 no.4
• /
• pp.94-104
• /
• 2012

Purpose: The purpose of this paper is to report the effect of Korean medicine treatments on a female and a male patient who were infertile and failed to in vitro fertilization(IVF) treatment. Methods: The patients are a 30-year-old female who had been diagnosed with the ovulation dysfunction caused by the depression of ovarian function, and a 33-year-old male who had been diagnosed with sperm defect. The female patient alone underwent Korean medicine treatment after the eighth failure in IVF treatment and the male patient received Korean medicine treatment along with his wife after he failed in the first IVF treatment. The Korean medicine applied include herb-medicine administration, acupuncture, moxibustion, Jwa-Hun, and Du-Han-Jok-Yeol treatment. Results: Through Korean medicine treatment the female patient had succeeded in the IVF treatment and she also recovered completely from allergic rhinitis. The male patient whose wife also received Korean medicine treatment with him got his wife pregnant naturally. Conclusions: This result concludes that Korean medicine treatment is effective in infertile patients. Therefore, there needs to be more of subsequent research on infertile patients that is based on Korean medicine treatment.

• Journal of Animal Reproduction and Biotechnology
• /
• v.34 no.1
• /
• pp.35-39
• /
• 2019

This study aimed to recover the ovarian function through in vitro culture of preantral follicles from aged mice. First, we isolated the preantral follicles from ovaries of sixty-seven-week old B6D2F1 mice with decreased fecundity to know how many follicles were present in them, which was 6 preantral follicles including 2 primary, 2 early secondary and late secondary follicles from 8 aged mice. It was confirmed that a few follicles (~2) were present in aged mice through histological analysis compared to adult mice as control. The 9 days of in vitro culture of preantal follicles showed in vitro growth and induced maturation after treatment with hCG (2.5 IU/mL) and EGF (5 ng/mL). Cumulus cells in the cumulus-oocyte complexes (COCs) were removed using hyaluronidase and oocytes at the germinal vesicle (GV) and GV breakdown (GVBD) were obtained from preantral follicle culture of aged mice in vitro. In conclusion, these observations demonstrated that there still were a few preantral follicles in the ovaries of 67 week-old mice, which we were able to culture in vitro and oocytes were obtained from them. This study proposed an in vitro culture system using preantral follicle as a therapeutic strategy for fertility preservation in humans for assisted reproductive medicine.