• Korean Journal of Animal Reproduction
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• v.16 no.1
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• pp.39-46
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• 1992

In viro fertilizatin is very important in both human clinical practice and animal breeding. However, the success rate of in vitro fertilization is not high. The purpose of this study ws to determine wheter in the vitro fertilization and culture of porcine oocyte using a hydrogel chamber were possible or not. Hydrogel chambers were made of polymerized 2-hydroxyethyl methacrylate. Matured follicular oocytes in Waymouth's medium and T L Hepes medium, tubal oocytes, and preincubated sperm in M199 medium were treansferred into the lumen of the hydrogel chambers. The chambers containing porcine oocytes and spermatozoa implanted into the mouse peritioneal cavity, and ova were examined after the recovery of the chambers at 84 hours after preservation start. The result was shown that fertilization and culture of porcine oocytes were successfully achieved inside of the hydrogel chamber.

• Korean Journal of Animal Reproduction
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• v.16 no.4
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• pp.335-340
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• 1993

The development of efficient method for the production of transgenic mice has been investigated in our laboratory. This study was conducted to develop the artificial insemination in the mouse. Spermatozoa were collected from the cauda epididymis of ICR males(age:12~15 weeks, Body weight : 30g) and artificially inseminated into the intrauterine via cervix of hormone-primed ICR females(age: 6~8 weeks, body weight: 25g) using the capillary tube, 200~300 $\mu$m in inner diameter. The effect of concentration of sperm(80$\times$104, 40$\times$104, 20$\times$104, 10$\times$104, 5$\times$104, 3$\times$104, 1$\times$104/20${mu}ell$. The artificial insemination was succeeded but fertilization rate was very low(5~15%) compared to the natural mating and 59 normal youngs born from 60 females. Therefore, our findings suggest that it is possible to produce the great number of mice from the same orgin of male by artificial inseminatin. However, the lower pregnancy rate has to be solved to used broadly the artificial insemination in mouse.

• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.83-83
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• 2002

This study was to compare the characteristics of parthenogenetic mES (P-mES) cells and in vitro fertilization mES cells. Mouse oocytes were recovered from superovulated 4wks hybrid F1 (C57BL/6xCBA/N) female mice. The oocytes were treated with 7% ethanol for 5 min and 5 ㎍/㎖ cytochalasin-B for 4 h. For IVF, the oocytes were inseminated with epididymal sperm of hybrid Fl male mice (1×10/sup 6//㎖). IVF and parthenogenetic embryos were cultured in M16 medium for 4 days. Cell number count in blastocysts was carried out differential labelling using propidium iodide (red) and bisbenzimide(blue). (omitted)

• Korean Journal of Animal Reproduction
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• v.15 no.3
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• pp.225-231
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• 1991

These experiments were carried out ot detect autoantiboies to zona pellucida in sera from infertile women using indirect ELISA and IFA and to investigate their effect on in vitro fertilization of mouse ova. In inidirect ELISA test, 12 of 116(10.3%) serum samples form infertile women gave positive reaction whereas all of 16 samples from fertile women and men were negative. Furthermore, in indirect IFA test, 17 of 116 (14.7%) serum samples from infertile women gave positive fluorescence whereas all of control sera were negative fluorescence. The fertilization rates(15.9%) of mouse eggs treated with positive sera were significantly lower than those(51.9%＋71.2%) autoantibodies to zonapellucida are responsible for infertility in unexplained infertile women, presumably by perventing sperm attachment and penetration.

• Journal of Embryo Transfer
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• v.27 no.4
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• pp.205-209
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• 2012

Transgenic rats and mice are useful experimental animal models for medical research including human disease model studies. Somatic cell nuclear transfer (SCNT) technology is successfully applied in most mammalian species including cattle, sheep, pig and mouse. SCNT is also considered to increase the efficacy of transgenic/knockout mouse and rat production. However, in the area of reproductive biotechnology, the rodent model is inadequate because of technical obstacles in manipulating the oocytes including intracytoplasmic sperm injection and SCNT. In particular, success of rat SCNT is very limited so far. In this review, the history of rodent cloning is described.

• Development and Reproduction
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• v.16 no.4
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• pp.363-370
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• 2012

The outer dense fiber 2 (ODF2) protein is an important component of sperm tail outer dense fiber and localizes at the centrosome. It has been reported that the RO072 ES cell derived homozygote knock out of ODF2 results in an embryonic lethal phenotype, and XL169 ES cell derived heterozygote knock out causes severe defects in sperm tail development. The ODF2s splicing variant, Cenexin1, possesses a C-terminal extension, and the phosphorylation of serine 796 residue in an extended C-terminal is responsible for Plk1 binding. Cenexin1 assembles ninein and causes ciliogenesis in early stages of the cell cycle in a Plk1-independent manner. Alternatively, in the late stages of the cell cycle, G2/M phase, Cenexin1 binds to Plk1 and results in proper mitotic progression. In this study, to identify the in vivo function of Plk1 binding to phosphorylated Cenexin1 S796 residue, and to understand the in vivo functional differences between ODF2 and Cenexin1, we generated ODF2/Cenexin1 S796A/Cenexin1 WT expressing transgenic mice in a RO072 ES cell derived $ODF2^{+/-}$ knock out background. We observed a severe defect of sperm tail development by ectopic expression of Cenexin1 S796A mutant and no phenotypic differences between the ectopic expression of ODF2/Cenexin1 WT in $ODF2^{+/-}$ background and in normal wild type mice.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.1
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• pp.73-85
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• 2020

A high-cholesterol diet can reduce male fertility. However, it is not known whether a high-cholesterol diet can regulate the expression of genes involved in sperm maturation and sperm fertilizing ability. Quercetin, a natural product, is known to have cytoprotective effects by regulating lipid metabolism in various cell types. This study aimed to confirm the expression of genes involved in sperm maturation in the testes of mice fed a high-cholesterol diet and to determine whether quercetin can reverse the genetic regulation of cholesterol. Mice were divided into groups fed a normal chow diet and a high-cholesterol diet. Mice fed the high-cholesterol diet were dose-dependently supplemented with quercetin for 6 weeks. Investigations using quantitative PCR and in situ hybridization revealed that the high-cholesterol diet alters the expression of genes associated with sperm maturation in the testes of mice, and this was reversed with the supplementation of quercetin. In addition, the high-cholesterol diet regulated the expression of genes related to lipid metabolism in the liver of mice. Under a high-cholesterol diet, quercetin can improve male fertility by regulating the expression of genes involved in sperm maturation.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.96-96
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• 2003

Nek2 (NIMA-related protein) is a mammalian cell cycle-regulated kinase that involves in chromosome condensation and centrosome regulation and NuMA (nuclear mitotic apparatus protein) is involved in spindle assembly during a cell cycle. The cellular distribution and organization of the centrosomal components is completely unknown during fertilization and embryonic development. We examined distribution of two well-known centrosomal proteins, Nek2 and NuMA in mouse gametes and embryos to get an insight in the reorganization of centrosomal proteins during germ cell development and early fertilization. Spermatogenic cells, gametes, and embryos were analyzed with anti-Nek2 or -NuMA antibodies by immunological assay, RT-PCR, and overexpression through gene transfection. Mitotically or meiotically active spermatogenic cells were intensively stained with these antibodies in both centrosomes and cytoplasm, whereas the oocytes showed different staining patterns depending on the meiotic stages. During maturation, GV, GVBD, and MI stage were clearly stained with NuMA antibody in the nucleus or cytoplasm at MII. Also, Nek2 was detectable in cytoplasm as scattered spots or chromosome associated at MII. In early developmental embryo, NuMA was detected in nucleus of each blastomere, while Nek2 was detected in cytoplasm. In contrast to previously reported results, Nek2 and NuMA were detected in both decondensing head, and the centriole of demembranated and decondensed sperm or whole body of trypsin-treated sperm for Nek2. During meiotic progress in oocytes, transcripts levels were the highest in MI stage and then downregulated in MII. Also, it shows dramatically change in early developmental embryos, firstly, it was increased until 4 cell stage and reduced in 8 cell stage, and finally, transcript levels were upregulated until blastoscyst. This finding suggests that cnetrosomal component may play an important role in reorganizing of functional centrosome during fertilization process and subsequent development.

• The Korean Journal of Zoology
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• v.33 no.1
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• pp.70-77
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• 1990

The change of phosphatase activities of the epididymal spermatozoa has been examined during epididymal maturation in mouse. The quantitative analysis of phQsphatase activities have been carried out using the method modified by Emst(1975). The results of experiment were summarized as the followings. Total protein of the caput epididyrnal spermatozoa(CPS) was measured as 59.1 $\pm$8.4(mg/10 9 spermatozoa), and that of the cauda epididymal spermatozoa(CDS) was 14.0$\pm$12.3(mg/10 9 spermatozoa). When phosphatase activities of the CDS in basic reaction medium were 29.2% in alkaline phosphatase, 44.9% in ATPse and 53.8% in acid phosphatase. The activities were eminently decreased in all CDS in contrast to those of CPS. The alkaline phosphatase and ATPase activities of K+ -dependent were decreased in CDS when compared with caput epididymal spermatozoa, and alkaline phosphatase, ATPase and acid phosphatase activities of $Ca^2$+ -dependent were increased in homogenized spermatozoa when compared with intact spermatozoa. From these results, it may be concluded that the decrease of phosphatases activities in spermatozoa during epididymal maturation may play some significant roles in acquiring fertilizing capability.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.213-213
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• 2004

Exogeneous DNA can reproducibly be delivered by co-injected spermatozoa and this transgenesis method is very efficient protocol. But, mosaic patterns of transgenic embryos and offspring were shown frequently. Whole blastomere integration is important in transgenic animal economics. (omitted)