• The Journal of Korean Obstetrics and Gynecology
• /
• v.27 no.1
• /
• pp.120-139
• /
• 2014

Objectives: The tendency of experimental studies on the male infertility was analyzed through focusing on domestic theses for a degree to indicate the hereafter direction for its study in oriental herbal medicine. Methods: 35 domestic theses for a degree published after 2003 were analyzed. And theses were classified by year, study design, degree and subjects. Results: The followings are the results of this study. 1. 35 theses were reviewed. 11 master dissertations and 24 doctorate dissertations 2. In the annual publishing tendency, the number of theses began to increase from 2003 and the theses have been published actively from 2005. 3. Classified by study design, 19 in vitro & in vivo and 16 in vitro experimental studies were performed. 4. In the medication and prescription, the studies with single medication were 32, and prescription were 3. In the studies with single medication, four studies on Ginseng Radix (人蔘) were the most. 5. In theses related to single medication, BoYang-Yak was most prefered, followed BoYeum-Yak, BoKi-Yak, SabJung-ChukNyo-JiDae-Yak. 6. Analysed 35 theses by subject, concentrational experiments were 23, periodical experiment were 3, concentrational & periodical experiment was 1. 7. In theses related to herb medication, the themes were Body & testicular weight, sperm parameters, testosterone level, CREM mRNA level & CREM protein, seminiferous tubule and antioxidant activity. 8. Related to herb medication's antioxidant activity efficacy were 7 theses. Conclusions: If there are more diverse studies on medication, prescription, external treatment methods and experiment methods in the future, this will be very helpful for the clinical treatment of male infertility.

• The Korean Journal of Malacology
• /
• v.29 no.4
• /
• pp.263-272
• /
• 2013

The development of swift's scallop Chlamys swiftii, reared in the laboratory, has been examined through the investigation of morphological characteristics in fertilized egg, larvae and juvenile. Eggs were fertilized with a dilute sperm solution to improve the survival of fertilized eggs. Developing larvae were maintained at a temperature of $16{\pm}0.5^{\circ}C$ and salinity concentration of 33 ppt. We have investigated the fertilization rates and egg number spawned at several stimulating conditions such as sunlight exposure, air dry, seawater temperature rise ($5^{\circ}C$) and seawater temperature rise ($5^{\circ}C$) after exposure of air dry for spawning induction of swift's scallop Chamys swiftii. Stimulation treated with sunlight exposure and seawater temperature rise ($5^{\circ}C$) have shown the spawning number of 700,000-900,000 and 700,000-800,000 per individual, respectively while stimulation treated with seawater temperature rise ($5^{\circ}C$) after exposure of air dry have shown the high spawning number of 1,000,000-1,500,000 per individual. Survival rate of D-shaped larvae of swift's scallop put into the different seawater temperatures of $8^{\circ}C$, $12^{\circ}C$, $16^{\circ}C$, $20^{\circ}C$ and $24^{\circ}C$ has been 4.1%, 11.6%, 32.7%, 18.6% and 3.2%, respectively. Fertilized eggs with the diameter of $72{\mu}m$ developed into trochophore larvae of $103{\pm}3.8{\mu}m$ shortly after 35 hours and to D larvae of $129{\pm}10.4{\mu}m$ shell length within 72 hours. It took 336 hours to become initial Umbo-stage larva of $145{\pm}16.8{\mu}m$ shell length. Post larvae, which have been $197{\pm}13.6{\mu}m$ shell length, spontaneously have settled in the attachment substances. It have required 528 hours from fertilized eggs to early attached juvenile to become initial juvenile size of $245{\pm}15.8{\mu}m$ shell length.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.33 no.5
• /
• pp.431-438
• /
• 2000

To clarify the annual reproductive cycle in a rockfish, Sebastes schlegeli, monthly changes in gonadosomatic index (GSI), hepatosomatic index (HSI) and histological feature of gonads and plasma levels of sex steroid hormones ($estradiol-l7{\beta},\;17{\alpha},\;20{\beta}-dihydroxy-4-pregnen-3-one,\;testosterone\;and\;11-ketotestosterone$) were investigated. The annual reproductive cycle in females could be divided into 5 periods as follows: 1) recovery period (June to September): serum level of $estradiol-l7{\beta}$ increased gradually; 2) vitellogenesis period (Septemer to february) : vitellogenic oocytes were obsewed, GSI sustained high value, and serum level of $estradiol-l7{\beta}$ increased; 3) gestation period (February-April): developing larva showed in the ovary, and serum levels of $17{\alpha},\;20{\beta}-dihydroxy-4-pregnen-3-one$ and testosterone increased; 4) partrition period (April to May) : larva were delivered, and value of GSI and serum levels of hormones decreased rapidly; 5) resting period (May to June) : value of GSI and serum levels of $estradiol-l7{\beta}$ and testosterone remained low. The annual reproductive cycle in males could be divided into 6 periods; 1) early maturation period (April to June): value of GSI and serum levels of hormones incresed gradually, cyst of spermatogonia incresed in number, and a small number of cyst of spermatocyte was observed; 2) mid-maturation perid (June to September); value of GSI and serum levels of hormones increased, and germ cells in many cysts were undergoing active sperrnatogenesis; 3) late maturation period (September to November) : value of GSI and serum levels of hormones remained high and spermatozoa were released into the lumina of the seminal lobules; 3) spermatozoa dischaging period (Nobember to December) : the lumina of the seminal lobules were enlarged and filled with mature spermatozoa; 4) degeneration period (December to Februauy)i value of GSI decresed and cyst of spermatocyte were decresed in number; 5) resting period (December to April) : no histological changes of testes were observed, and value of GSI and serum levels of hormones remained low. In November, the lumina of the seminal lobules were filled with mature spermatozoa and sperm masses were present in the ovarian cavity. Thus, copulation in this species occurred in November and December.

• Korean Journal of Animal Reproduction
• /
• v.22 no.1
• /
• pp.1-9
• /
• 1998

This study was carried out to confirm whether the developmental capacity of bovine mature oocytes frozen ultra-rapidly using electron microscopic(EM) grids and EFS30 can be obtained, and whether the cryoprotectants and the freezing method used in this study effect detrimentally to the bovine oocytes by indirect immunocytochemistry. As freezing solution, we used EFS30 which consisted of 30% ethylene glycol, 0.5 M sucrose, 18% ficoll and 10% FBS added in D-PBS. The results obtained in this experiment were summarized as follows: When the effects of cryoprotectant and freezing procedure on the microtuble, micrfilament and chromatin morphology of oocytes were evaluated using indirect immunocytochemistry, the results of freezing as well as exposure group were not different with that of the control oocytes. When the fertilization abnormality after ultrarapid freezing of bovine mature oocytes was examined by Hoechst staining, the rates of total penetration(96.7, 9.0%), normal two pronuclei formation(74.6, 68.9%) and mean number of sperm / oocyte(1.50, 1.44) were not different between control and freezing group. In addition, when the developmental capacity of frozen-thawed of oocytes(85.5%) was survived, 74.5% of them were cleaved and 31.4% of cleaved embryos were developed to blastocyst. These data were similar to those of the control(76.0%, 34.6%) and exposure(74.5%, 33.0%) except survival rates. Also, when the total cell number of blastocysts produced from the each treatment at day after IVF was examined by hoechst staining, there were not different among groups. There results demonstrate that developmental capacity of frozen-thawed bovine mature oocytes can be successfully obtained by ultra-rapid freezing method using EM grid and EFS30 solution.

• Journal of Embryo Transfer
• /
• v.20 no.3
• /
• pp.289-295
• /
• 2005

This study was performed to investigate the relationship between Heat shock protein 70 (HSP70) gene polymorphism and in vitro fertilization(IVF) embryo development in the pigs. The single strand conformation polymorphism(SSCP) genotypes from HSP70 K1, K3 and K4 PCR products were detected different patterns. In cleavage rate of oocyte fertilized in vitro, HSP70 K1-AA genotype($73.1\%$) and K1-AB genotype($62.3\%$) showed significantly higher oocyte cleavage rate than HSP70 K1-BB genotype($49.3\%$)(p<0.05). And HSP70 K3-AA genotype ($72.4\%$) and K3-AB($62.2\%$) also showed significantly higher oocyte cleavage rate than HSP70 K3-BB genotype($49.1\%$)(p<0.05). The IVF embryo development of 2-cell stage according to HSP70 genotypes of sperm and pig breeds also showed a significant difference. The number of embryos developed to 2-cell stage in Landrace(28.8) and Duroc(29.8) were significantly higher than in Yorkshire(10.9)(p<0.05). And also HSP70 K4-AB genotype group(29.6) higher than HSP70 K4-AA genotype group(10.6)(p<0.05). However, the number of embryos developed to blastocyst stage did not showed significant differences among breeds as well as HSP70 genotypes. These resrults suggest that in vitro development in porcine early embryos may be affected by HSP70 genotypes and breeds.

• Clinical and Experimental Reproductive Medicine
• /
• v.25 no.1
• /
• pp.87-92
• /
• 1998

This study was carried out to investigate in vitro/in vivo development of vitrified mouse oocytes. Mouse oocytes were vitrified using EFS30, 35 and 40 (30, 35 and 40% ethylene glycol, 18% ficoll and 0.5 M sucrose in M2 medium). After being exposed or vitrified-thawed, oocytes of normal morphology were inseminated in vitro by $1-2\times10^6/ml$ of epididymal sperm. The rates of fertilization, in vitro/in vivo development and cell number (inner cell mass and tropectoderm cell) of blastocysts in each treatment group were examined. The results obtained in these experiments were summarized as follows: The cleavage rates were obtained in EFS35 containing 35% ethylene glycol higher than in EFS30 and EFS40. The development rate of vitrified-thawed oocytes to two-cell stage after in vitro fertilization (51.1%) was significantly different compared to that of exposed to vitrification solution without cooling (60.0%) and control (68.2%) (p<0.05). However, there were no differences in the blastocyst formation from the cleaved embryos among groups (75.0, 73.3 and 80.0%). Also, the mean number of cells per blastocysts of vitrified group $(92.5{\pm}2.9)$ was similar to that of the exposed $(98.5{\pm}5.3)$ and control $(100.9{\pm}4.8)$. In vivo development of the blastocysts derived from vitrified-thawed oocytes resulted in fetal development (50.7%) and implantation rates (80.0%) which are very similar to those of control (58.2, 78.2%). These results suggest that mouse oocytes could be cryopreserved using vitrification solution (EFS35) based on ethylene glycol.

• Reproductive and Developmental Biology
• /
• v.28 no.2
• /
• pp.119-126
• /
• 2004

The present study was carried out to examine the effect of four different media (NCSU (North Carolina State University)-23, PZM (Porcine Zygotes Medium)-3, PZM-4 and TCM (Tissue Culture Medium)-l99) and two oxygen concentrations (39 , 5% $O_2$, 5% $CO_2$ and 90% $N_2$, 5% $CO_2$ in air) on in vitro production of porcine IVM/IVF embryos. The results were summarized as follows: The rates of GVBD and nuclear maturations were not significantly different (p>0.05) for 44 hours of culture with four media in two oxygen concentrations. The rates of polyspermy, penetrated sperm(s) and male and female prouclei formation were not significantly different (p>0.05). among four media in two oxygen concentrations. The cleavage rates were not significantly different (p>0.05) among four media in two oxygen concentrations. At day 7 under gas atmosphere of 5% $O_2$, 5% $CO_2$ and 90% $N_2$, the blastocyst formation was significantly higher (p<0.05) in PZM-3 (19.9$\pm$2.4) than other media. Also, NCSU-23 medium gave high rate of blastocyst formation at day 7 under gas atmosphere of 5% $CO_2$ in air (p<0.05). Based on the result of differential staining of porcine blastocyst at dat 7, inner cell mass cell and total cell numbers were not significantly different (p>0.05) among four media in two oxygen concentrations. However, the observed total cell number was higher in PZM-3 medium (36.8$\pm$6.5) than other madia. In conclusion, these results suggested that in vitro production of porcine embryos in PZM-3 medium under a gas atmosphere of 5% $O_2$, 5% $CO_2$ and 90% $N_2$ was effective on the blastocyst formation rate and total blastocyst cell number.

• Journal of Embryo Transfer
• /
• v.19 no.3
• /
• pp.245-255
• /
• 2004

In recent years, an increasing number of studies on pig in vitro maturation(IVM) and in vitro fertilization(IVF) have been separated. the wide range of new technologies, including that in applied molecular genetics, has increased this interest. the production of viable porcine embryos in vitro is a prerequisites for the successful production of transgenic pigs to date. The efficiency of IVM/IVF techniques in the porcine is lower than that obtained in other species such as cattle and mouse. The several problems are generally thought to be the cause of poor results: the low rate of MPN formation derived from inadequate IVM of oocytes, the high incidence of polyspermy after IVF and cell blocking at 4 cell during embryos culture. For there reasons overcoming, many studies have been conducted to improve in vitro embryo-genic competence of oocytes. In the last several years, many maturation culture media have been evaluated and various exogenous factors such as hormones and grows factors have been tested to improve the efficiency of porcine in vitro system. In the study several antioxidants have been examined to improve in vitro fertilization and development of porcine oocytes. In this study, several antioxidants were examined to determine the effects on the development of oocytes to the cleavage, morula and blastocyst stage when added at the maturation(IVM) or in vitro fertilization(IVF) or in vitro culture(IVC) of porcine embryos. Porcine oocytes were matured, fertilized and embryos were cultured in defind conditioned medium in vitro with or without supplementation with the antioxidents of cysteine, catalase and glutathione. 1. Significant improvement of blastocyst rate (27.2％ versus 15.4％, p<0.05) were achieved when catalase(500U/$m\ell$) were added to TCM-199 medium and morula rate(72.0％ versus 53.9％, p<0.05) were significantly higher when glutathione(1.0mM/$m\ell$) were added to TCM-199 medium than those of control. 2. In mTBM medium for oocytes fertilization, the addition of cysteine, catalase and glutathione had no positive effect on embryonic development. glutathione had no positive effect on embryonic development. In conclusion, this study shows that addition of catalase, gluththione during IVM improved the rate of porcine embryo development.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.15 no.3
• /
• pp.241-250
• /
• 1982

The structure of gonads, gametogenesis and reproductive cycle of the cockle, Fulvia mutice, were studied mainly by histological observation. The materials were monthly sampled in the southern area of Yeosu from October 1980 to September 1981. F. mutica was monoecious. The gonads were situated between the liver tissues and the outer fibronuscular layers compacted by the connective tissue fibers and muscle fibers beneath the outermost layer of simple cuboidal epithelium. The gonad was composed of a number of the ovarian sacs and the testicular tubules which form the tubular structure. Testicular tubules in the mature stage sometimes contained 'testis-ova' The undifferentiated mesenchymal tissues and the eosinophilic cells were abundantly distributed on the germinal epithelium in the early development stage. With the further development of the ovary and testis, these tissues and cells gradually disapprared. The undifferentiated mesenchymal tissues and the eosinophilic cells are related to the growing of the oocytes and spermatocytes . Early multiplicating oogonium was about $10{\mu}m$ in diameter. As the oocytes grow to $27-34\times50-58{\mu}m$ by increasing cytoplasm, the oocytes connected to the basement membrane by their egg-stalks. The ripe eggs were about $60{\mu}m$ in diameter and they were surrounded by gelatinous membrane. Most male germ cells in mature stage were transformed into the spermatozoa and they formed the sperm bundles. After spawning, undischarged ripe eggs and spermatozoa remained in the ovarian sac and the testicular tubule respectively for some time, then they finally degenerated. Especially the early spent ovarian sacs in May did not contract significantly and then they took part in the secondary maturation within two or three months during the summer season. The monthly changes of the fatness well agreed with the reproductive cycle. The reproductive cycle of F. mutica could be classified into six successive stages : multiplicative, growing, mature, spent, degenerative and recovery stage. It seems that the spawning season is closely rotated to the water temperature, and the spawning occurs from May to October at about $20^{\circ}C$ in water temperature. The peak spawning seasons appeared twice a year between June and July and in September. Acknowledgement The authors wish to express their gratitude to Dr. Kim, In Bae, Dr. Chun, Seh Kyu and Dr. Yoo, Sung Kyoo of National Fisheries University of Busan and Mr. Min, Byoung Seo of National fisheries Research and Development Agency for their critical reading of the manu script.

• Journal of Embryo Transfer
• /
• v.26 no.1
• /
• pp.19-25
• /
• 2011

This study was carried out to evaluate the effect of early pregnant cow as donor for Ovum Pick-Up (OPU) derived oocyte aspiration and embryo production in Holstein heifers. Four non-pregnant and 2 pregnant Holstein heifers were used as donor and then carried out total 17 OPU session for 10 weeks (2 times per week). Recovered cumulus-oocyte-complexes (COCs) were classified into 4 grade by oocyte cytoplasm and cumulus cells and matured in vitro in TCM-199 supplemented with 10% FBS, 10 mg/ml FSH and 1 mg/ml estradiol in 5% $CO_2$ and over 99% humidity for 24 h. After 24 h co-incubation with post-thaw sperm, the presumed zygotes were cultured in CR1aa medium with 4 mg/ml BSA for 3 days and then changed CR1aa medium with 10% of FBS for another 3~4 days. The Mean number of aspirated follicles and collected oocytes in the early stage pregnant and non-pregnant heifers were $13.0{\pm}4.3$ and $10.6{\pm}3.9$, $5.4{\pm}3.4$ and $7.7{\pm}3.6$ per session, respectively. Rate of collected oocyte from aspirated follicles were 59.2% and 50.5%, respectively. The average number of good quality oocytes (Grade I and II) in the early stage pregnant and non-pregnant heifers was $3.7{\pm}2.7$ and $4.9{\pm}2.6$ (Mean${\pm}$SD). Cleavage and blastocyst developmental rates in Grade I and II were 22.2% and 25.5%, and then $1.7{\pm}0.9$ and $1.4{\pm}1.1$ blastocyst per session, respectively. In conclusion, OPU technology can be used in early stage pregnant and non-pregnant heifers without any problem and so applied OPU derived embryo production to maximize the ability of genetically valuable females.