• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.69-69
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• 2003

Although effect of TGF$\beta$$_1$ on preimplantation embryo development was reported at mice, little information relevant to this subject is known in bovine. The objectives of this study were to investigate TGF$\beta$$_1$, and TGF$\beta$$_1$ receptors type I and II expression, known as important factors in the embryo development, at unfertilized oocytes and fertilized embryos that will be used as basic data to be compared to NT embryos. We postulated that TGF$\beta$$_1$ may have a beneficial effect on the preimplantation embryo and show different expression patterns as embryo stages change. We have used immunocytochemistry to investigate the presence in unfertilized oocytes and preimplantation embryos of TGF$\beta$$_1$ and the essential components of the TGF$\beta$$_1$ signalling pathway, TGF$\beta$$_1$ receptors type I and II. We found that both receptors, as well as TGF$\beta$$_1$, were present in the unfertilized oocytes. This indicates that TGF$\beta$$_1$, is a maternally expressed protein. At the morulae and blastocyst stages the TGF$\beta$$_1$ receptor type II was not present, but the TGF$\beta$$_1$ receptor type I was present at both stages and we can confirm the TGF$\beta$$_1$ expression of high level at 8-cell stage. These findings support our hypothesis that the TGF$\beta$$_1$, and TGF$\beta$$_1$ receptors may interact with the oocyte and preimplantation embryo, and that TGF$\beta$$_1$ signalling may be important for the development of the oocyte and the preimplahtation embryo.

• 한국발생생물학회지:발생과생식
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• 제12권1호
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• pp.19-30
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• 2008

수정란이 포배로 분화하는 것은 착상을 통하여 개체 발생이 성립되는 포유동물의 발생에 있어서 핵심적인 현상이다.. 초기 배아 발생 시기동안 배아는 생존을 위한 에너지원을 공급받아야 한다. 포유동물의 난자는 보통의 경우 난자 형성 동안 많은 양의 에니지원을 세포질에 비축하지 않기 때문에 발생 동안 수란관과 자궁으로부터 물질대사와 관련돼 여러 물질, 특히 에너지원을 획득해야 한다. 탄수화물은 착상전 배아의 주 에너지원으로 알려져 있다. 포도당, 젖산염, 피르브산염은 착상 전배아 배양액에서 없어서는 않될 성분으로, 초기 배아는 그 발생 단계에 따라 이들 물질에 대한 선호도를 각기 다르게 갖고 있다. 포도당수송체(glucose transporter)와 수소이온-단당류 동향수송체($H^+$-monocarboxylate cotransporter)는 탄수화물을 수송하는 주된 매개자로 이들의 발현 수준은 일차적으로 내인성 또는 인슐린이나 포도당과 같은 외인성 요인에 동시적으로 조절을 받는다. 비록 1960년대 이후 화학적으로 규명된 BWW와 같은 배양액을 이용하여 수정란이 성공적으로 포배로 발생되고 이식 후 정상적인 새끼가 태어났어도, 발생조절에 있어서 이들 탄수화물 물질대사 산물의 역할은 잘 알려져 있지 않다. 포도당은 밀착이 진행되는 상실배에서 물질대사 관련 효소와 수송체의 발현을 조절하고, 포배강 형성에 필요로 하는 에너지를 생산하는데 관련된 것으로 인식되고 있다. 다른 한편으로 cytokine은 배아에서 탄수화물의 대사율, 그리고 물질대사율 조절을 통하여 배아 발생을 조절할 수 있는 것으로 제안되어 왔다. 또한, 근래 들어 본인 등은 젖산염이 착상 전 배아의 발생을 조절할 수 있는 물질임을 밝히고 있다. 이러한 결과들은 탄수화물의 물질대사물이 초기 배아 시기에 에너지원으로서 뿐만이 아니라 생합성 경로 및 다른 조절경로에 참여하고 있음을 의미한다. 따라서 초기 배아 발생 동안 탄수화물 대사와 대사물질은 에너지원으로서 뿐만이 아니라 수정란이 착상할 수 있는 능력을 갖춘 포배로 발생하는 것을 조절하는 조절물질로 그 중요성이 있다.

• Animal cells and systems
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• 제2권1호
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• pp.99-106
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• 1998

In this study, we have examined the role of cAMP in gap junctional communication (GJC) in preimplantation mouse embryos. GJC was monitored by Lucifer Yellow (LY) injected into one blastomere of compacted embryos. The speed of GJC was defined as the time taken for the last blastomere of the embryo to become visibly fluorescent. The median time for 8-cell embrvos (140 sec) was similar to that for 16-cell (135 sec). To determine whether cAMP and cAMP-dependent protein kinase (PKA) are involved in the regulation of GJC, the effects of PKA inhibitor (H8) and cAMP analogues (Rp-cAMP and 8-Br-cAMP) on dye transfer between blastomeres of compacted embryos were examined. Some of the embryos treated with either H8 or Rp-cAMP failed to transfer LY to all blastomeres within 10 min. In contrast, 8-Br-cAMP speeded up fluorescent dye transfer. The median time to fill all blastomeres with LY was 140 sec in untreated controls and 90 sec in siblings treated with 8-Br-cAMP. Inhibition of PKA by H8 or Rp-cAMP induced delay or arrest in embryo development after compaction, but the increase of intracellular cAMP showed no effect. These findings suggest that GJC in preimplantation mouse embryos is regulated by cAMP-PKA pathway and transient interference by PKA inhibitors induces the developmental delay beyond compaction.

• 한국가축번식학회지
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• 제16권4호
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• pp.341-346
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• 1993

The development of isolated blastomeres from mammalian preimplantation embryos has been basically studied for the multiplication of embryos from superior animals. Therefore, this study was investigated the effect of co-culture with mouse fetal fibroblast cells(MFFC) on in vitro development of blastomeres from mouse preimplantation embryos. Mature female ICR mice were treated with hormone to induce superovulation and embryos were collected at each 2, 4, and 8-cell stage. Then, after removing zona pellucida with protease, blastomeres were isolated by micropipetting, or reconstituted with different stage blastomere, and incubated for 72 hrs either in T6 or TCM199 or on the monolayer of MFFC, which was prepared with fibroblast cells from 14∼14 day mouse fetus. After incubation, we examined their development rates every day and the nuclei numbers of each blastocyst by Hoechst-33342 staining. In the development rates of blastomeres, there were no significant differences between media but the higher rateswere found in the monolayer of MFFC, regardless of reconsititution. In addition, blastomeres cultured with MFFC had slightly greater number of nuclei than those cultured in single media. Generally, the higher development rates of blastomeres were found from earlier stage embryos than the later ones, regardless of culture conditions. Reconsitituted blastomeres had more nuclei but did not show the higher development rates, compared to the single blastomeres. Taken together, our results suggest that co-culture with MFFC have a beneficial effect on the in vitro development of blastomeres from mouse embryos.

• 한국수정란이식학회지
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• 제25권3호
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• pp.179-187
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• 2010

Embryonic compaction is essential for normal preimplantation development in mammals. The present study was to investigate the effects of compaction patterns on developmental competence of pig embryos. The proportion of blastocyst formation derived from compacted morula was higher than those of compacting and pre-compacting morula (P<0.01). Nuclei numbers of inner cell mass (ICM), trophectoderm (TE), and total of blastocysts derived from compacted group were also superior to those of compacting and pre-compacting groups (P<0.05). Then, compaction patterns, developmental ability and structural integrity were compared between mono- and poly-spermic embryos. The rate of compacted morula in mono-spermic embryos was higher than that of poly-spermic embryos (P<0.05). Especially, the rate of blastocyst formation derived from compacted embryos in mono-spermic embryo group was higher than that of poly-spermic embryo group (P<0.05), although no difference was detected between the two groups in the structural integrity. Finally, we confirmed that beta-catenin was differentially expressed according to compaction patterns in morula and blastocyst stage embryos. In conclusion, our results suggest that the compaction patterns during preimplantation development play a direct role in developmetal competence and quality of pig embryos.

• 한국가축번식학회지
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• 제12권1호
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• pp.31-36
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• 1988

There studies were conducted using inbred ICR mice to examine the sex of preimplantation mouse embryo. The morphological normality of mice embryos treated with the culture medium containing rat H-Y antiserum(10%, v/v) plus complement(20%,v/v) was observed and also the sexing of embryos was investigated by chromosomal analysis. The results obtained were summarized as follows: 1. The viability of preimplantation mouse embryos, which were incubated in vitro with different media condition, was scored 68.9-85.5% in control group. However, 151 embryos normally developed up to blastocyst and 160 embryos were retarded growth or destroyed out of total 311 embryos treated in the medium containing H-Y antiserum(10%, v/v) plus complement(20%,v/v). 2. H-Y antiserum was prepared from inb red rats (Wistar and Donryu strain) with different immunization times (4, 5 and 6th) to examine the specific titer of embryos by the number of immunization. Precentage of normally developed embryos incubated either in the medium containing the antiserum of Wistar plus complement or Donryu plus complement was revealed 50.9, 47.4 and 50.0% (4, 5 and 6th immunization and 47.8, 41.2 and 48.7%, respectively. 3. Twenty two females and five males were identified out of fourty-eight normally developed embryos incubated in the medium containing H-Y antiserum plus complement by chromosomal analysis.

• Clinical and Experimental Reproductive Medicine
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• 제46권4호
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• pp.206-210
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• 2019

Mucopolysaccharidosis type II (MPS II) is a rare X-linked recessive lysosomal storage disease caused by mutation of the iduronate-2-sulfatase gene. The mutation results in iduronate-2-sulfatase deficiency, which causes the progressive accumulation of heparan sulfate and dermatan sulfate in cellular lysosomes. The phenotype, age of onset, and symptoms of MPS II vary; accordingly, the disease can be classified into either the early-onset type or the late-onset type, depending on the age of onset and the severity of the symptoms. In patients with severe MPS II, symptoms typically first appear between 2 and 5 years of age. Patients with severe MPS II usually die in the second decade of life although some patients with less severe disease have survived into their fifth or sixth decade. Here, we report the establishment of a preimplantation genetic diagnosis (PGD) strategy using multiplex nested polymerase chain reaction, direct sequencing, and linkage analysis. Unaffected embryos were selected via the diagnosis of a single blastomere, and a healthy boy was delivered by a female carrier of MPS II. This is the first successful application of PGD in a patient with MPS II in Korea.

• Clinical and Experimental Reproductive Medicine
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• 제42권1호
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• pp.1-7
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• 2015

Stress coping mechanisms are critical to minimize or overcome damage caused by ever changing environmental conditions. They are designed to promote cell survival. The unfolded protein response (UPR) pathway is mobilized in response to the accumulation of unfolded proteins, ultimately in order to regain endoplasmic reticulum (ER) homeostasis. Various elements of coping responses to ER stress including Perk, Ask1, Bip, Chop, Gadd34, Ire1, Atf4, Atf6, and Xbp1 have been identified and were found to be inducible in oocytes and preimplantation embryos, suggesting that, as a normal part of the cellular adaptive mechanism, these coping responses, including the UPR, play a pivotal role in the development of preimplantation embryos. As such, the UPR-associated molecules and pathways may become useful markers for the potential diagnosis of stress conditions for preimplantation embryos. After implantation, ER stress-induced coping responses become physiologically important for a normal decidual response, placentation, and early organogenesis. Attenuation of ER stress coping responses by tauroursodeoxycholate and salubrinal was effective for prevention of cell death of cultured embryos. Further elucidation of new and relevant ER stress coping responses in periimplantation embryos might contribute to a comprehensive understanding of the regulation of normal development of embryonic development and potentiation of embryonic development in vitro.

• Molecules and Cells
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• 제20권3호
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• pp.423-428
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• 2005

Immediately after fertilization, a chromatin remodeling process in the oocyte cytoplasm extracts protamine molecules from the sperm-derived DNA and loads histones onto it. We examined how the histone H3-lysine 9 methylation system is established on the remodeled sperm chromatin in mice. We found that the paternal pronucleus was not stained for dimethylated H3-K9 (H3-$m_2K9$) during pronucleus development, while the maternal genome stained intensively. Such H3-$m_2K9$ asymmetry between the parental pronuclei was independent of $HP1{\beta}$ localization and, much like DNA methylation, was preserved to the two-cell stage when the nucleus appeared to be compartmentalized for H3-$m_2K9$. A conspicuous increase in H3-$m_2K9$ level was observed at the four-cell stage, and then the level was maintained without a visible change up to the blastocyst stage. The behavior of H3-$m_2K9$ was very similar, but not identical, to that of 5-methylcytosine during preimplantation development, suggesting that there is some connection between methylation of histone and of DNA in early mouse development.

• Clinical and Experimental Reproductive Medicine
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• 제27권3호
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• pp.307-311
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• 2000

Objective: To report the successful delivery in a patient of recurrent spontaneous abortion caused by chromosomal abnormality. Material and Method: Case report. Results: Twelve oocytes were obtained by in vitro fertilization. Of eleven oocytes fertilized, two embryos turned out to be normal by using fluorescent in situ hybridization on blastomere biopsy. The patient succeeded in pregnancy and the result of amniocentesis was found to be normal. She delivered the healthy female baby by cesarean section. Conclusions: The successful delivery is possible in recurrent spontaneous abortion related with reciprocal translocation by using preimplantation genetic diagnosis.