• The Korean Journal of Zoology
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• v.39 no.2
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• pp.147-158
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• 1996

Since the floating goby, Choenogobius onnuloris, has intricate and diverse morphological variations, allozymic analysis at 25 loci was carried out for their populations in Korea to clarify its taxonomic status. A genetic assay carried out revealed that the floating gobies were clearly divided into three genetic groups (Tvpe-A, Tvpe-B and Tvpe-C) in Korea. Alternative alleles were fated at six loci (Aco, 6pgd, Ldh-1, Got-1, Gpf, Gp3l. Some loci had considerable heterogeneitv among three types and no evidence of gene exchange in slunpatric populations was found from statistical analyses. The genetic similarity (Rogers'S) among three types was lower than 0.80 and divergent time estimate indicates that they were speciated during 1.2-1.8 million year before present (WBP). ASso, these three types of C. unnuluris were distinguished morpholosicallv from each other by several characters such as band ornamentations. In conclusion, the evidences presented here support recognition of three types of Chuenogobius annularis as typical discrete species.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.54 no.2
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• pp.170-179
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• 2021

This study was conducted to identify taxonomic differences in the characteristics of Acheilognathus majusculus and A. yamatsutae during their initial life history via an interspecific hybridization experiment. Hatching time required 36 h for MY and 49 h for YM at 21.5℃, showing a significant difference of 13 h between the hybrids. The hatching rates of the cross-bred eggs were 30% for cross MY (A. majusculus♀×A. yamatsutae♂) and 40% for cross YM (A. yamatsutae♀×A. majusculus♂). The hatching larvae size was total length 3.13-3.43 mm in MY and total length 3.89-4.22 mm in YM, which was larger in YM. The hybridization test between A. yamatsutae and A. majusculus that live in the same water stream confirmed that no interspecific reproductive isolation occurred.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.55 no.2
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• pp.137-145
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• 2022

This study was conducted to identify the taxonomic differences in the characteristics of Rhodeus ocellatus and Acheilognathus yamatsutae during their early life history via an interspecific hybridization experiment. Results showed that the hatching rates of the cross-bred eggs were 60% for cross OY (R. ocellatus ♀×A. yamatsutae ♂) and 70% for cross YO (A. yamatsutae ♀×R. ocellatus ♂). The dorsal fin rays of YO were similar to those of R. ocellatus. The hybridization test of A. yamatsutae and R. ocellatus living in the same water stream revealed that no interspecies reproductive isolation occurred. OY and YO showed similar maternal and paternal characteristic. The juvenile of OY did not resemble both paternal and maternal fin rays, indicating a difference. The hatching larvae of the hybrids showed the morphological characteristics of their parents but varied in terms of the shapes of their parts.

• Clinical and Experimental Reproductive Medicine
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• v.33 no.4
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• pp.265-272
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• 2006

Objective: This study was carried out to evaluate the effect of the isolation methods of inner cell mass from mouse blastocyst, types of feeder cells and treatment time of mitomycin C on the formation rate of ICM colony. Methods: The inner cells were isolated by conventional immunosurgery, partial trophoblast dissection with syringe needles and whole blastocyst co-culture method. Commercially available STO and primary cultured mouse embryonic fibroblast (pMEF) feeder cells were used, and mitomycin C was treated for 1, 2 or 3 hours, respectively. The formation rate of ICM colony was observed after isolation of ICM and culture of ICM on the feeder cells for 7 days. Result: The ICM colony formation rate on STO were significantly higher in partial trophoblast dissection group (58%) than that in immunosurgery (12%) or whole blastocyst culture (16%) group (p<0.05). The formation rate on pMEF feeder layer was higher in partial trophoblast dissection (88%) and whole blastocyst culture (82%) group than that in immunosurgery (16%) group (p<0.05). When mitomycin C treated to pMEF for 2 hours, the formation rate of 88% was significantly higher than those of other conditions. Conclusion: Above results showed that the efficient isolation method of ICM from blastocyst was the partial trophoblast dissection and the appropriate treatment time of mitomycin C was 2 hours. However, the subculture of ICM colony and characterization of stem cells should be carried out to confirm the efficacy of the partial trophoblast dissection method.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.4
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• pp.430-436
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• 2010

The objective of the present studies was to develop and validate a system for isolation, purification and extended culture of pigment-producing cells in alpaca skin (melanocytes) responsible for coat color and to determine the effect of alpha melanocyte stimulating hormone treatment on mRNA expression for the melanocortin 1 receptor, a key gene involved in coat color regulation in other species. Skin punch biopsies were harvested from the dorsal region of 1-3 yr old alpacas and three different enzyme digestion methods were evaluated for effects on yield of viable cells and attachment in vitro. Greatest cell yields and attachment were obtained following dispersion with dispase II relative to trypsin and trypsin-EDTA treatment. Culture of cells in medium supplemented with basic fibroblast growth factor, bovine pituitary extract, hydrocortisone, insulin, 12-O-tetradecanolphorbol-13-acetate and cholera toxin yielded highly pure populations of melanocytes by passage 3 as confirmed by detection of tyrosinase activity and immunocytochemical localization of melanocyte markers including tyrosinase, S-100 and micropthalmia-associated transcription factor. Abundance of mRNA for tyrosinase, a key enzyme in melanocyte pigment production, was maintained through 10 passages showing preservation of melanocyte phenotypic characteristics with extended culture. To determine hormonal responsiveness of cultured melanocytes and investigate regulation of melanocortin 1 receptor expression, cultured melanocytes were treated with increasing concentrations of ${\alpha}$-melanocyte stimulating hormone. Treatment with ${\alpha}$-melanocyte stimulating hormone increased melanocortin receptor 1 mRNA in a dose dependent fashion. The results demonstrated culture of pure populations of alpaca melanocytes to 10 passages and illustrate the potential utility of such cells for studies of intrinsic and extrinsic regulation of genes controlling pigmentation and coat color in fiber-producing species.

• Clinical and Experimental Reproductive Medicine
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• v.46 no.4
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• pp.152-165
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• 2019

Objective: This study aimed to examine the effect of vitrification on apoptosis and survival in human preantral follicles after thawing. Methods: This experimental study was conducted at an acute tertiary care hospital from March 2012 to April 2013. Ovaries were sliced into 5 × 5 × 1-mm pieces and divided into the following three groups: preantral follicle isolation, ovarian tissue vitrification-warming followed by follicle isolation, and immunohistochemistry of fresh ovarian tissue. For statistical analyses, the Student t-test, chi-square test, Kruskal-Wallis test, and Kaplan-Meier survival analysis were used. Results: A total of 161 preantral follicles (70% secondary) were collected from ovarian cortex tissue of six women between 30 and 37 years of age who underwent oophorectomy due to cervical cancer or breast cancer. There were no significant differences in the follicular morphology of fresh preantral follicles and vitrified follicles after thawing. The mean Fas ligand (FasL) mRNA expression level was 0.43 ± 0.20 (relative to β-actin) in fresh preantral follicles versus 0.51 ± 0.20 in vitrified follicles (p= 0.22). The mean caspase-3 mRNA expression level in fresh preantral follicles was 0.56 ± 0.49 vs. 0.27 ± 0.21 in vitrified follicles (p= 0.233). One vitrified-thawed secondary follicle grew and developed to an antral follicle within 6 days of culture. Conclusion: Vitrification did not affect preantral follicle morphology or mRNA expression of the apoptosis markers FasL and caspase-3. Further studies are required to establish whether vitrification affects the outcomes of in vitro culture and the maturation of preantral follicles.

• Journal of Veterinary Clinics
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• v.34 no.2
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• pp.140-145
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• 2017

The goal of this study was to investigate risk factors associated with porcine reproductive and respiratory syndrome (PRRS) in pig farms in the Republic of Korea using logistic regression and a multilevel model. A cross-sectional study was applied to 305 pig farms with a questionnaire-based interview by veterinarians between March 2014 and February 2015. The questionnaire comprised eight categories: proximity to neighbors, disinfection, visitors, vehicles, insecticides, wild animals, gilts, and feeding. In total, 61 questions in eight categories related to pig farm biosecurity were investigated. Farms were classified as PRRS stable or unstable based on the results of an antibody test and PCR. For univariate analysis, keeping production records with computers (OR = 0.283, 95% CI = 0.056 - 1.425), accredited farm with no use of antibiotics (OR = 0.412, 95% CI = 0.134 - 1.269), reviewing health record of semen prior to purchasing (OR = 0.492, 95% CI = 0.152 - 1.589), complete isolation of runt pigs (OR = 0.264, 95% CI = 0.084 - 0.829), compulsory registering for visitors (OR = 0.424, 95% CI = 0.111 - 1.612), keeping records of insecticide history (OR = 0.406, 95% CI = 0.089 - 1.846), routine on-farm monitoring by veterinarians (OR = 0.314, 95% CI = 0.069 - 1.423), and use of on-farm checklist for biosecurity monitoring (OR = 0.313, 95% CI = 0.063 - 1.553) were found to decrease the probability of PRRS infection. Multivariate and multilevel analysis revealed only two factors, complete isolation of runt pigs (OR = 0.165, 95% CI = 0.045 - 0.602 and OR = 0.208, 95% CI = 0.055 - 0.782) and compulsory registering for visitors (OR = 0.106, 95% CI = 0.017 - 0.655 and OR = 0.119, 95% CI = 0.017 - 0.809) were found to decrease the probability of PRRS infection. The intracluster correlation coefficient of a province for multilevel model was 0.05. The results of this study might facilitate biosecurity measures for individual farms to reduce the probability of PRRS infection.

• Korean Journal of Veterinary Research
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• v.43 no.3
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• pp.463-469
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• 2003

Eight nursery to grower pigs exhibiting weight loss and sudden death were diagnosed as postweaning multisystemic wasting syndrome (PMWS) based on the results of gross findings, histopathology, immunohistochemistry, fluorescent antibody test, virus isolation, PCR, serology, and electron microscopy. Groosly, the pigs had a rough hair coats and were severely emaciated. And moot lymph nodes were pale and enlarged. Lungs were not fully collapsed and exhibited 10 to 40% pale red cranioventral consolidation. Histopathologically, typical lymphohistiocytic interstitial to bronchointerstitial pneumonia, chronic lymphadenitis, severe lymphoid depletion, and basophilic intracytoplasmic inclusions were noted in the most lymphoid tissues. Porcine circovirus panicles were observed in the inguinal lymph node of the pigs by electron microscopy. Porcine circovirus type 2 (PCV2) antigens or viral DNAs were detected in the lesions of all pigs using immunohistochemistry or PCR. Two PCV2 were isolated from a homogenate of pooled lung and lymph node in 2 of the 5 pigs. Additionally, antigens of porcine reproductive and respiratory syndrome virus (PRRSV) and Hemophilus (H.) parasuis were also detected by immunofluorescent antibody test. Serologically, 55% of randomly selected sows and fattening pigs was serum antibody positive to PCV2 by an indirect fluorescent antibody (IFA) test and approximately 18 % of animals in the herd were serologically pooitive by the ELISA kit for PRRSV. To our knowledge, this is the first report of PMWS co-infected with PCV-2, PRRS, and H. parasuis in Korea.

• Clinical and Experimental Reproductive Medicine
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• v.45 no.1
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• pp.17-24
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• 2018

Objective: To investigate sperm chromatin/DNA integrity, global DNA methylation, and DNMT mRNA transcription in men with oligoasthenoteratozoospermia (OAT) compared with normozoospermic men. Methods: Semen samples from 32 OAT patients who comprised the case group and 32 normozoospermic men who comprised the control group were isolated and purified using a standard gradient isolation procedure according to World Health Organization criteria. DNMT1, DNMT3A, and DNMT3B transcripts were then compared between groups using real-time quantitative reverse-transcription polymerase chain reaction. Global DNA methylation in sperm was determined by an enzyme-linked immunosorbent assay. Protamine deficiency and the proportion of apoptotic spermatozoa were evaluated using chromomycin A3 (CMA3), aniline blue (AB), and toluidine blue (TB) staining, as well as the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The p-values < 0.05 were considered to indicate statistical significance. Results: Significantly higher proportions of AB+, TB+, CMA3+, and TUNEL+ spermatozoa, as well as DNMT3A and DNMT3B transcription, were found in the OAT group. Positive correlations were detected between sperm parameters, DNA/chromatin damage, and DNMT3A and DNMT3B transcripts. Global DNA methylation was significantly higher in the OAT patients and had a significant correlation with abnormal results of all sperm chromatin integrity tests, but was not associated with DNMT1, DNMT3A, or DNMT3B expression. Conclusion: Oligoasthenoteratozoospermic men showed abnormal sperm parameters, abnormal chromatin/DNA integrity, and a higher global DNA methylation rate, as well as overexpression of DNMT mRNA.

• Clinical and Experimental Reproductive Medicine
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• v.38 no.2
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• pp.68-74
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• 2011

Objective: Previously, we found that oocyte specific homeobox (Obox) 4 plays significant role in completion of meiosis specifically at meiosis I-meiosis II (MI-MII) transition. The purpose of this study was to determine the mechanism of action of $Obox4$ in oocyte maturation by evaluating downstream signal networking. Methods: The $Obox4$ dsRNA was prepared by $in$ $vitro$ transcription and microinjected into the cytoplasm of germinal vesicle oocytes followed by $in$ $vitro$ maturation in the presence or absence of 0.2 mM 3-isobutyl-1-metyl-xanthine. Total RNA was extracted from 200 oocytes of each group using a PicoPure RNA isolation kit then amplified two-rounds. The probe hybridization and data analysis were used by Affymetrix Gene-Chip$^{(R)}$ Mouse Genome 430 2.0 array and GenPlex 3.0 (ISTECH, Korea) software, respectively. Results: Total 424 genes were up (n=80) and down (n=344) regulated after $Obox4$ RNA interference (RNAi). Genes mainly related to metabolic pathways and mitogen-activated protein kinase (MAPK) signaling pathway was changed. Among the protein kinase C (PKC) isoforms, PKC-alpha, beta, gamma were down-regulated and especially the MAPK signaling pathway PKC-gamma was dramatically decreased by $Obox4$ RNAi. In the cell cycle pathway, we evaluated the expression of genes involved in regulation of chromosome separation, and found that these genes were down-regulated. It may cause the aberrant chromosome segregation during MI-MII transition. Conclusion: From the results of this study, it is concluded that $Obox4$ is important upstream regulator of the PKC and anaphase-promoting complex action for maintaining intact germinal vesicle.