• Clinical and Experimental Reproductive Medicine
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• 제28권1호
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• pp.13-24
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• 2001

Objective: In muscle and neuronal cells, calcium channels have been classified by electrophysiological and pharmacological properties into (1) voltage-dependent $Ca^{2+}$-channel (1) P/Q-type $Ca^{2+}$-channel (2) N-type $Ca^{2+}$-channel (3) L-type $Ca^{2+}$-channel (4) T-type $Ca^{2+}$-channel (5) R-type $Ca^{2+}$-channel. The present study was done in order to investigate whether there is any difference in $Ca^{2+}$-channel distribution between activated and normally fertilized embryos. Methods: The immunocytochemical method was used to identify the existence of voltage-dependent $Ca^{2+}$-channels in parthenogenetically activated 2-cell embryos by ethanol and $SrCl_2$ treatment. These 2-cell embryos were obtained by exposure to 6% ethanol for 6 min and to 10 mM $SrCl_2$ for 2h. Results: P/Q-type $Ca^{2+}$-channels and L-type $Ca^{2+}$-channels have been identified. Whereas, three type of $Ca^{2+}$-channel P/Q-type, N-type, L-type have been identified in 2-cell embryos fertilized in vivo. Conclusion: Activation by ethanol was faster than those by $SrCl_2$. However, there was difference in DAB staining of the embryos between ethanol and $SrCl_2$ treatment (87.7% and 54.1 %). Intensity of staining was also different between ethanol- and $SrCl_2$-treated group. However, it has not been known why there was some difference in DAB staining and staining intensity in the present study.

• Journal of Periodontal and Implant Science
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• 제41권4호
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• pp.192-200
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• 2011

Purpose: The aim of this study is to compare the gene expression profile in mesenchymal stem cells derived from dental tissues and bone marrow for characterization of dental stem cells. Methods: We employed GeneChip analysis to the expression levels of approximately 32,321 kinds of transcripts in 5 samples of bone-marrow-derived mesenchymal stem cells (BMSCs) (n=1), periodontal ligament stem cells (PDLSCs) (n=2), and dental pulp stem cells (DPSCs) (n=2). Each cell was sorted by a FACS Vantage Sorter using immunocytochemical staining of the early mesenchymal stem cell surface marker STRO-1 before the microarray analysis. Results: We identified 379 up-regulated and 133 down-regulated transcripts in BMSCs, 68 up-regulated and 64 down-regulated transcripts in PDLSCs, and 218 up-regulated and 231 down-regulated transcripts in DPSCs. In addition, anatomical structure development and anatomical structure morphogenesis gene ontology (GO) terms were over-represented in all three different mesenchymal stem cells and GO terms related to blood vessels, and neurons were over-represented only in DPSCs. Conclusions: This study demonstrated the genome-wide gene expression patterns of STRO-$1^+$ mesenchymal stem cells derived from dental tissues and bone marrow. The differences among the expression profiles of BMSCs, PDLSCs, and DPSCs were shown, and 999 candidate genes were found to be definitely up- or down-regulated. In addition, GOstat analyses of regulated gene products provided over-represented GO classes. These data provide a first step for discovering molecules key to the characteristics of dental stem cells.

• 대한바이러스학회지
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• 제26권2호
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• pp.151-162
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• 1996

Pseudorabies is caused by Pseudorabies virus (PRV: Aujeszky's disease virus) of Herpesviridae that is characterized by 100 to 150nm in size with a linear double-stranded DNA molecule with of approximately $90{\times}10^6Da$. This disease affects most of domestic animals such as swine, cattle, dog, sheep, cat, chicken, etc. causing high mortality and economic losses. In swine, young piglets show high mortality and pregnant sows, reproductive failures. However the adult swine reveals no clinical signs in general. But they become a carrier state and play an important role for propagation of the disease. In this study, the nucleotide sequence of major casid protein gene of PRV, Yangsan strain isolated from the diseased swine in Korea was analyzed, and the recombinant MCP was produced by expression of the MCP gene in Sf-9 cell using baculovirus transfer vector system. As result, in BamHI digestion, MCP gene locus of PRV YS strain showed different from that of Indiana S strain. The patterns of enzyme mapping were also found to be unidentical each other. The sequence of the MCP gene partially analyzed showed 98.09% identity to Indiana S strain. The expression of MCP in Sf-9 cell cotransfected by pVLMCP-44 baculovirus expression vector was characterized by Southern blot hybridization, immunofluoresent and immunocytochemical tests, SDS-PAGE and Western blotting. The rMCP with M.W. 142kDa was most effectively expressed in Sf-9 cells at the 3-4th days post inoculation of the recombinant baculovirus by 2 moi.

• 한국안광학회지
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• 제19권2호
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• pp.271-277
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• 2014

목적: 본 연구의 목적은 브라질산 주머니쥐(Monodelphis domestica) 망막에서의 calretinin 면역반응성을 연구하는데에 있다. 칼슘결합단백질 calretinin은 칼슘매개-신호전달에서 중요한 역할을 한다고 알려져 있다. 방법: 표준 면역조직화학법을 이용하여 브라질산 주머니쥐 망막을 대상으로 실험하였다. 결과: 브라질산 주머니쥐 망막에서 calretinin은 일부의 수평세포, AII 무축삭세포, 일부의 신경절세포에서 면역반응성을 보였다. 특히, calretinin 면역반응성을 가지는 모든 AII 무축삭세포는 또 다른 칼슘결합단백질인 parvalbumin을 발현하였다. 결론: 다른 포유류 망막과 다를 바 없이, 브라질산 주머니쥐 망막에서도 AII 무축삭세포에서 calretinin 면역반응성이 관찰되었다. 이를 통해 calretinin이 브라질산 주머니쥐 망막에서도 AII 무축삭세포의 marker로 이용될 수 있다는 점을 확인하였다.

• Applied Microscopy
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• 제42권3호
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• pp.115-123
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• 2012

Human placenta extract (hPE) has therapeutic potential against certain diseases such as burn injury, liver cirrhosis and chronic wound through stimulating tissue repair processes. However, the effects of hPE on liver regeneration in animals are unknown. This study investigated the effect of hPE on the expression of proliferating cell nuclear antigen (PCNA) during liver regeneration induced by partial hepatectomy (PH) in rats. The activities of AST, ALT and ALP increased during a few days after PH. A high level of ALP was particularly seen at day 3 in the control group. All the levels of experimental groups were normalized by day 5 after PH. On immunohistochemistry, the expression of PCNA increased at the early days, showed a peak at day 3 after PH. The PCNA staining was more obvious in the experimental group over the whole period. By western blotting, PCNA seemed to be more strongly expressed in the hPE injected group in the early stage and fell to almost undetectable levels at day 7. On immunocytochemical observations, the number of PCNA-gold particles in the nuclei at day 1 of the hPE treated groups was more than those of the untreated groups. The results suggest that hPE could accelerate liver regeneration induced by PH involving the expression of PCNA in rats.

• 한국발생생물학회지:발생과생식
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• 제4권1호
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• pp.7-12
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• 2000

수정에 의한 배 발생은 정자가 난자 내로 침입하여 정자와 난자의 반수체 핵질이 융합되고 이어 유사분열로 이어지는 과정에서 시작된다. 하지만 수정 및 초기 배 발생 동안 자웅 핵질과 난 세포질 구성 요소 상호간의 작용기전에 관해서는 명확히 알려져 있지 않은 부분이 많다. 수정보조기법인 세포질 내 정자 직접 주입법의 개발은 남성불임치료에 혁신적인 기술로 자리잡고 있을 뿐만 아니라 포유동물의 수정과정을 이해하는데 많은 도움을 주고 있다. 핵치환에 의한 복제동물 생산기법도 분화된 핵이 난 세포질 내에서 재 분화 (reprogramming)하여 발생하는 유일한 과정으로 세포질 구성요소들의 상호작용과 발생 조절 기능을 이해하는데 도움을 준다. 최근 몇 년간 돼지 난자 세포질에 정자 및 원형정자 직접주입, 세포질 이식, 세포질 융합 및 핵치환 한 후 난자의 발생과정을 간접 면역형광 분석법과 주사 전자현미경으로 조사하였다. 이러한 연구를 통해 체외수정, 세포질 이식 및 정자직접 주입법 등과 같은 임상치료기술 과 핵치환에 의한 복제동물생산 기법의 개선에 필요한 기초자료를 얻을 수 있었고, 포유동물 난자의 후생적 발생과정 (epigenesis)에 관해 공부할 수 있었다.

• 대한세포병리학회지
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• 제9권1호
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• pp.1-14
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• 1998

The cytologic distinction of carcinoma cells from reactive mesothelial cells can be difficult, especially in specimens containing abundant reactive mesotheilal cells and inflammatory cells with scant carcinoma cells. This study evaluates the usefulness of mucin and immunocytochemistry for discrimination between reactive mesotheilal cells and carcinoma cells, and sensitivity and specificity of these stains for the detection of metastatic carcinoma in serous effusions. Immunocytochemical panel including mucin cytochemistry with the periodic acid-Schiff(PAS) reaction after or without diastase digestion was undertaken on 127 serous effusion specimens with histologically confirmed diagnoses. The specimens including cell smears and cell blocks were stained with PAS and antibodies to carcinoembryonic antigen(CEA), epithelial membrane antigen(EMA), cytokeratln(CK), and vimentin. The sensitivities of these stains for metastatic carcinoma(127 cases) were 49%(46/94) in PAS, 48%(60/124) in CEA, 89%(97/109) in EMA, 88%(93/106) in CK, and 25%(20/81) in vimentin. The sensitivities of stains for reactive mesothelial cells(36 cases) were 19%(7/36) in EMA, 78%(28/36) in CK, and 75%(27/36) in vimentin. The PAS and CEA stains were not reacted with all cases of benign reactive serous effusions containing abundant reactive mesothelial cells. The specificities of stains for metastatic carcinoma(127 cases) were 100% in PAS, 100% in CEA, 81% in EMA, 22% in CK, and 25% in vimentin. The optimal combination of stains for use in a panel was PAS and CEA. Combined results from these two stains yielded an advanced sensitivity of 8% in PAS and 4% in CEA for metastatic carcinoma. EMA was also cosiderably useful for identification of carcinoma cells. CK and vimentin were not suitable for distinguishing between reactive mesothelial cells and carcinoma cells.

• 한국가축번식학회지
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• 제27권4호
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• pp.309-315
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• 2003

DNA methylation at CpG sites, which is a epigenetic modification, is associated with gene expression without change of DNA sequences. During early mouse embryogenesis, dynamic changes of DNA methylation occur. In this study, DNA methylation patterns of porcine embryos produced in vivo and in vitro were examined at various developmental stages by the immunocytochemical staining method. Interestingly, active demethylation was not observed on the paternal pronucleus of porcine zygotes. However, differences were detected in the passive demethylation process between in vivo and in vitro embryos. There was no change in the DNA methylation state until the blastocyst stage of in vivo embryos, whereas partial demethylation was observed in several blastomeres from a 4 cell stage to a morula stage of in vitro embryos. The whole genome of inner cell mass (ICM) and trophectoderm (TE) cells in porcine blastocysts were evenly methylated without de novo methylation. Our findings demonstrate that genome-wide demethylation does not occur in pig embryos during preimplantation development unlike murine and bovine embryos. It indicates that the machinery regulating epigenetic reprogramming may be different between species.

• 한국안광학회지
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• 제16권1호
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• pp.91-95
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• 2011

목적: 본 연구는 토끼 망막에서 특정 신경 세포 염색이 가능한 염료를 조사하였다. 방법: Rhodamine B를 토끼의 안구내 유리체강에 주입하였다. 24시간 후, 망막을 분리하여 현미경으로 염색 상태를 관찰하였다. 염색 된 세포는 그 종류를 알아보기 위하여 면역조직화학적방법을 실시하였다. 결과: 망막 내핵층 중간 위치에서 선명하게 염색된 핵들이 관찰되었다. 염색된 세포의 분포도와 수가 뮬러세포와 비슷한 양상을 보였다. 뮬러세포인지 확인하기 위하여 vimentin 항체를 사용하였다. Rhodamine B에 의해 염색된 핵들은 vimentin 항체로 감싸져 있어 뮬러세포임을 확인하였다. 결론: Rhodamine B를 살아있는 망막에 넣을 경우 토끼 망막에 신경교세포의 특이적 염색이 나타났다.

• 한국안광학회지
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• 제18권2호
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• pp.187-191
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• 2013

목적: 한국관박쥐의 망막에서 원뿔세포의 middle/long(ML) opsin cone photoreceptors의 분포를 분석하여 박쥐의 시각계를 이해하고자 하였다. 방법: 표준면역세포화학법을 이용하여 성체 한국관박쥐의 망막을 대상으로 조사하였다. 결과: 4 개체의 망막 전체에서 추정된 ML opsin은 $27,336{\pm}2,130$개였으며, 평균밀도는 $7,854{\pm}268cells/mm^2$이었다. S opsin은 외핵층에 위치한 세포외절에서 일부 면역반응성을 보였다. 결론: ML opsin의 조직화된 분포와 S opsin의 발현 결과는 한국관박쥐가 밝은 빛에도 반응하며 색을 구별할 수 있는 기능을 가지고 있다는 것을 알 수 있다.