• Applied Microscopy
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• 제25권3호
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• pp.1-19
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• 1995

The present study was performed to clarify the effect of vagus nerve stimulation on the enterochromaffin(EC) cells in the body of the stomach, the first part of the duodenum and the ceceum of rats by using routine electron microscopy and immunogold labelling. The changes in the ultrastructure and in the labelling density of the gold particles of the EC cells were investigated after vagus nerve stimulation. The vagus nerve was electrically stimulated with a square wave pulse generator for a duration of 5 minutes each, a total of 8 times at 2 minute intervals. Immunogold labelling demonstrated that the epithelial serotonin immunoreactive cells of the gastrointestinal tract are EC cells containing characteristic pleomorphic granules. Immunocytochemically labelled gold particles were largely concentrated in the dense matrix of the granules of the EC cell, and the labelling density of the gold particles considerably increased after the vagus nerve stimulation. Except for a slight activation of Golgi complexes, no remarkable changes in the ultrastructures of the EC cells were noted after the vagus nerve stimulation. The above results suggest that vagus nerve stimulation may activate serotonin biosynthesis in EC cells.

• Journal of Microbiology and Biotechnology
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• 제6권6호
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• pp.468-473
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• 1996

The genome of tomato spotted wilt virus (TSWV) is composed of three RNA segments, S, M, and L RNA and the 5.0 kb M RNA encodes two glycoproteins Gl, G2 and NSm protein of unknown function. In an effort to investigate the function of the NSm protein, antibody was raised against NSm fusion protein overexpressed in Escherichia coli. This antibody was used to detect the NSm protein by using western blot analysis and electron microscopic observation after immunogold labelling. For the cloning of the NSm gene, total RNA extracted from a TSWV infected plant was used for cDNA synthesis and polymerase chain reaction (PCR) instead of going through time-consuming virus purification. A protein band specifically reacting to the NSm antibody was detected from TSWV inoculated plants. The NSm protein was detected in the cell wall fraction and in pellet from low speed centrifugation when the infected plant tissue was fractionated into 4 fractions. In the immuno-electron microscopic observation, gold particles were found around the plasmodesmata of infected plant tissue. These results suggest that the NSm protein of TSWV plays some role in cell-to-cell movement of this virus.

• 한국전자현미경학회:학술대회논문집
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• 한국현미경학회 1997년도 제28차 춘계학술대회
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• pp.14-14
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• 1997

• The Korean Journal of Physiology
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• 제27권2호
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• pp.123-138
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• 1993

The mechanism of insulin action to increase glucose transport is attributed to glucose transporter translocation from intracellular storage pools to the plasma membrane in insulin-sensitive cells. The present study was designed to visualize the redistribution of the glucose transporter by means of an immunogold labelling method. Our data clearly show that glucose transporter molecules were visible by this method. According to the method this distribution of glucose transporters between cell surface and intracellular pool was different in adipocytes. The glucose transporter molecules were randomly distributed at the cell surface whereas the molecules at LDM were farmed as clusters. By insulin treatment the number of homogeneous random particles increased at the cell surface whereas the cluster forms decreased at the intracellular storage pools. It suggests that the active molecules needed to be evenly distributed far effective function and that the inactive molecules in storage pools gathered and termed clusters until being transferred to the plasma membrane.

• Journal of the Korean Wood Science and Technology
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• 제24권1호
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• pp.68-74
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• 1996

Present work was undertaken to investigate the origin of milled wood lignin(MWL) in the wood cell wall using immunocytochemical techniques, which can provide the information on the localization of specific antigens(MWL in the present study) to be examined. Spruce MWL dissolved in DMSO and emulsified with Freund adjuvant was injected directly into the mouse spleen. The animals were boostered at two-week intervals after the initial immunization. Blood samples were purified in standard procedures. The characteristics of antibodies against MWL were tested by indirect ELISA. Visualization of MWL was carried out using conventional indirect immunogold-labelling methods on the ultrathin sections of spruce wood. Immuno-TEM observations showed that the immunogold probes were selectively attached to secondary cell walls of spruce wood. The most intense labelling was frequently observed in the S2 layer. In contrast, gold labelling in the lignin-rich regions, such as middle lamella and cell corner was not found. The immuno-TEM provides an indication that spruce MWL originates from the S2 layer.

• 한국패류학회지
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• 제15권2호
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• pp.81-92
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• 1999

The histochemical, cytochemical, and immunocytochemical investigations were conducted to find out the cellulase activity in the accessory glands of the digestive system of the oriental land snail Nesiohelix samarangae under the LM, SEM, and TEM. The cellulase activity was shown in the epithelium of th digestive gland by labelling with the immunogold (protein-A gold) particles. The epithelial cells showing the cellulase activity were Type 1 and Type 3 cells out of five types of the epithelial cells of the digestive gland. None of epithelial cells of the mucus gland and the salivary gland and the salivary gland were not labeled with the immunogold particles.

• 한국식물병리학회지
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• 제14권3호
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• pp.229-239
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• 1998

Antibodies were raised against fusion proteins of the N-terminus and a region containing the GDNQ (Gly-Asp-Asn-Gln) polymerase motif of the L (polymerase) protein of sonchus yellow net virus (SYNV). Immunoblot analyses using these antibodies revealed the presence of the L protein in purified SYNV preparations and in nuclear extracts from infected tobacco. The serological analyses and detection in a polyacrylamide gels suggested that the L protein is present in at least a 20 fold lower abundance than the G, N, M1 and M2 proteins, and has size corresponding to a molecular weight of over 200 kDa as predicted from nucleotide sequence data. Electron microscopy with gold-labelled antibodies was used to localize the N, M2, and G proteins of SYNV in thin sections of infected tissue. When sections of SYNV-infected tissue were treated with antisera against total SYNV proteins and N protein, gold label could be detected in both the viroplasms and in virus particles. With the anti-M2 protein antiserum, the gold label was strongly localized in the viroplasms but only limited labelling of the virus particle sonly. Limited labelling of the L protein was observed in the viroplasms and the virus particles, presumably because of the low abundance of L protein in the tissues.

• Fisheries and Aquatic Sciences
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• 제1권2호
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• pp.187-191
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• 1998

Chemotherapy can not be applied for the control of fish viral diseases because viruses depend on host machinery for their replication. Although new control strategies including vaccination are under development, avoidance of virus introduction by rapid and correct diagnosis is the best way of fish viral disease control. Although observation of virus particles with an electron microscope is an easy method for virus detection, it take a few days for the sample preparation. In order to shorten the sample preparation time, microwave radiation was applied in the procedure. With this method, 15 seconds was enough for fixation of virus infected fish samples or cultured cells inoculated with infectious hematopoietic necrosis virus, which takes 2-4 hours with routine methods. Also four minutes was enough for polymerization of embedding resin which takes 24-48 hours with routine methods. Samples prepared with microwave were good enough for direct electron microscopic observation and immunogold labeling assay.

• 한국패류학회지
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• 제14권2호
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• pp.149-159
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• 1998

In order to observe the anticellulolytic localization in the epithelia of the digestive tract such as esophagus, crop, and intestine of a Korean land snail N. samarangae, a cytochemical method and a immunogold labelling method were applied. For the cytochemical study on the cellulase activity, Benedict reaction method applied. And for the immunocytochemical study, the rabbit serum immunoglobuins (IgG) was obtained from the rabbits injected with cellulase which was extracted from body fluid of the snail. The digestive tract tissues of N. samarangae were fixed with 4% paraformaldehyde and 2% OsO4 and embedded in Lowicryl K4M at -40$^{\circ}C$ under UV light (360 nm). The thin sections were loaded on the nickel grids and stained with the serum IgG and protein A-gold complex (particle size: 10 nm). Observations were undertaken with transmission electron microscope (Jeol, JEM-1010). The epithelium of the digestive tract was consisted of five types of cells. In the cytochemical study, the reaction products were found along the periphery of the vacuoles derived from the Bebedict reaction. In the immunocytochemical study, the protein-A gold particles were selectively labelled in Type 1, Type 3 and Type 4 cells in intestinal tissue. membranes of rER, in the surrounding cytoplasm of the rER and secretory granules, and in the apical cytoplasm of the cells. On the material being secreted from the apical cytoplasm was also labelled with the immunogold particles. The all results obtained throughtout present study suggest that the intestinal epithelium of the snail N. samarangae seretes cellulase as one of digestive enzymes.

• Applied Microscopy
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• 제32권4호
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• pp.411-419
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• 2002

사람의 위암 조직을 미세구조와 metallothionein (MT)에 대한 면역 조직 및 세포화학적 방법으로 조사하였던 바, 다음과 같은 결과를 얻어냈다. 위암 세포들은 핵 세포질비가 정상세포에 비해 크고, 불규칙한 핵과 이질염색질의 분포가 증가하였으며, 세포질내에서 유리리보솜의 분포가 뚜렷이 증가하였다. 면역 조직 및 세포화학적 방법으로 MT의 발현을 조사하였던 결과, 이 단백질은 위암조직의 암세포에서 반응성이 높게 나타났으며, 주로 핵 부위에 집중되는 경향을 보였으며, 특히 이질염색질과 인 부위에서 면역 금입자들이 주로 분포하는 것으로 관찰되었다. 이러한 결과들은 위암 세포의 미세구조가 미분화세포들이 나타내는 일반적인 특징과 비교되었으며, 위암 세포에서 MT가 증가하는 현상은 이 단백질이 세포질에서 합성되어 핵내로 수송된 후, 세포 증식을 위한 전사과정에 관여할 것임을 시사하는 것이다.