• Journal of the Korean Wood Science and Technology
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• v.24 no.1
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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.

• The Korean Journal of Malacology
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• v.14 no.2
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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.

• Animal cells and systems
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• v.5 no.3
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• pp.263-266
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• 2001

Metallothionein (MT) is induced in the regenerating rat liver. We have investigated expression of MT gene by RT PCR as well as specific localization of MT by immunocytochemistry in regenerating rat liver after partial hepatectomy (PH). MT mRNA level started to increase from 1 h and reached the peak at 8 h after PH. The level decreased gradually by 24 h, and became similar to that of control group. In the immunocytochemical study, in all groups treated with primary antibody, immunogold particles indicating the presence of MT were evenly distributed throughout both cytoplasm and nucleus of the rat hepatocytes. Within the nucleus, the gold particles appeared to be intensely localized in the areas of euchromatin and nucleolus. Within the cytoplasm, gold particles did not seem to adhere to mitochondria or Iysosomes, but were freely distributed. However, rough endoplasmic reticulum was the obvious compartment on which the gold particles were localized. Time course of MT immunoreactivity revealed that distribution of gold particles in hepatocytes increased gradually by 24 h, and decreased at 48 h after PH. Briefly, PH resulted in the sharpest increase in the expression of MT mRNA at 8 h and in the immunoreactivity of MT at 24 h, respectively. It is suggested that the increase of MT mRNA expression, the intensity of immunoreactivity and the specific localization of MT may be associated with the compensatory cell proliferation followed by PH.

• Animal cells and systems
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• v.1 no.3
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• pp.507-513
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• 1997

Sequential observations of binding patterns and structural effects of Bacillus thuringiensis var. kurstaki were made on fat body tissue of the fall webworm, Hyphantria cunea Drury. Fat body was cultured in vitro in the presence of purified 62 kDa endotoxin and then examined for protein synthesis and the localization of membrane-bound toxin detected by an antibody against the 62 kDa endotoxin. Protein synthesis was mostly inhibited at concentrations of 15 ${\mu}$g/ml and higher. Immunocytochemical observations suggest that the toxin binds to all exposed basal lamina surrounding the fat body without apparent specificity. The cytopathic effect delectable by scanning electron microscope is disintegration rather than cell swelling. The basal lamina bound toxin was eventually detached from the fat body and followed by an extrusion of cell contents like lipid granules.

• The Korean Journal of Zoology
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• v.34 no.1
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• pp.31-43
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• 1991

In the present study, the localization of myosin and the pathway of myofibrilogenesis of skeletal muscle cells in culture were investigated by immunocytochemical methods including immunoelectron microscopy and hybridoma formation. There were manly free ribosomes in the cytosol of the myoblast. At 48 hr of the culture, the myofilaments started to form and at 72 hr began to assemble to form myoabrils. At this time the Z band appeared, but the sarcomeres did not link together. Aker 96 hr of culture, adjacent sarcomeres linked together and shotved the typical striation of the skeletal muscle fiber. Myosin was synthesized in free ribosomes at 24 hr of culture and localized at many myofilaments at 48 hr and assembled mvoabrils at 72 hr and at 120 hr of the culture, myosin was localized at thick filaments of the A band.

• Animal cells and systems
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• v.1 no.1
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• pp.93-98
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• 1997

The present study investigated the cellular localization of a-subunit of Gq (Gaq) protein in developing L8E63, rat skeletal muscle cell line. The colocalization of Gaq with actin cytoskeleton was demonstrated by double-labeling experiments. In mononucleated myoblasts, the immuno-fluorescence staining pattern of Gaq was almost identical with that of F-actin visualized with rhodamine-conjugated phalloidin. However, this colocalization of Gaq with cytoskeleton was not maintained in multinucleated myotubes. The staining pattern of Gaq in myotubes did not match with any specific subcellular structure, but appeared as a uniformly distributed diffuse staining throughout the whole cell surface. Interestingly, change in the expression level of Gaq was not detected during myoblast differentiation, suggesting that actin-associated Gaq protein might dissociate from the cytoskeleton as cells differentiate. Immunocytochemical experiments using specific antibodies directed against several G proteins indicated that the subcellular localizations of Gai1, Gai2, Gai3, and Gao were different from those obtained with Gaq.

• The Korean Journal of Zoology
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• v.36 no.1
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• pp.116-122
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• 1993

An immunocytochemical investigation has been carried out to localize serotoninimmunoreactive (5-HTi) neurons in suboesophageal ganglion of fifth instar lawn of cabbage worm Pieris rupae. The 285-HTi cell bodies were identified in the rind of suboesophaseal ganglion. The four 5-HTi cell bodies of them are 18rge in size (about 35 Um), while the remaining cell bodies are medium-sized (about 15 Uml. The 5-HTi nerve processes are abundantly located in central large neuropil, circumoesophageal connectives which join the suboesophaseal ganglion to the tritocerebrum of the brain, and connectives between the suboesophageal and the first thoracic ganglia. These results indicate that the 5-HTi nerve fibers, which constitute the central large neuropil, have structural connections with the above two connectives. Especially in central large neuropil, many 5-HTi nenre fibers form a large circular bundle, in which a 5-HTi nerve fiber bundle is crossing.

• Molecules and Cells
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• v.19 no.3
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• pp.408-417
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• 2005

Nitric oxide (NO) occurs in various types of cells in the central nervous system. We studied the distribution and morphology of neuronal nitric oxide synthase (NOS)-containing neurons in the visual cortex of mouse and rabbit with antibody immunocytochemistry. We also compared this labeling to that of calbindin D28K, calretinin, and parvalbumin. Staining for NOS was seen both in the specific layers and in selective cell types. The densest concentration of intense anti-NOS immunoreactive (IR) neurons was found in layer VI, while the weak anti-NOS-IR neurons were found in layer II/III in both animals. The NOS-IR neurons varied in morphology. The large majority of NOS-IR neurons were round or oval cells with many dendrites coursing in all directions. Two-color immunofluorescence revealed that only 16.7% of the NOS-IR cells were double-labeled with calbindin D28K in the mouse visual cortex, while more than half (51.7%) of the NOS-IR cells were double-labeled with calretinin and 25.0% of the NOS-IR cells were double-labeled with parvalbumin in mouse. By contrast, 92.4% of the NOS-IR neurons expressed calbindin D28K while only 2.5% of the NOS-IR neurons expressed calretinin in the rabbit visual cortex. In contrast with the mouse, none of the NOS-IR cells in the rabbit visual cortex were double-labeled with parvalbumin. The results indicate that neurons in the visual cortex of both animals express NOS in specific layers and cell types, which do not correlate with the expression of calbindin D28K, calretinin or parvalbumin between the two animals.

• Journal of Plant Biology
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• v.35 no.2
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• pp.99-106
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• 1992

The endosperm protein, vicilin, of ginseng (Panax ginseng C.A. Meyer) was purified by ammonium sulfate precipitaion, gel permeation and ion exchange column chromatography. Vicilin is a glycoprotein composed of 2 subunits with molecular masses of 55,000 (large subunit) and 44,000 (small subunit). The anti-vicilin antibody was raised in rabbit, and purified by DEAE Affi-Gel Blue affinity chromatography. The endosperm cells of the seed were reacted with this anti-vicilin antibody and colloidal gold conjugated secondary antibody. Gold particles were labelled on the elaborating granules of Golgi complex, electron-dense granules and protein bodies in the endosperm cells. These results indicated that the vicilin, which was synthesized in rough endoplasmic reticulum and transported to Golgi, was elaborated in saccules of the Golgi and then transported into protein bodies by electron-dense granules.anules.

• BMB Reports
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• v.50 no.1
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• pp.37-42
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• 2017

The tubby protein (Tub), a putative transcription factor, plays important roles in the maintenance and function of neuronal cells. A splicing defect-causing mutation in the 3'-end of the tubby gene, which is predicted to disrupt the carboxy-terminal region of the Tub protein, causes maturity-onset obesity, blindness, and deafness in mice. Although this pathological Tub mutation leads to a loss of function, the precise mechanism has not yet been investigated. Here, we found that the mutant Tub proteins were mostly localized to puncta found in the perinuclear region and that the C-terminus was important for its solubility. Immunocytochemical analysis revealed that puncta of mutant Tub co-localized with the aggresome. Moreover, whereas wild-type Tub was translocated to the nucleus by extracellular signaling, the mutant forms failed to undergo such translocation. Taken together, our results suggest that the malfunctions of the Tub mutant are caused by its misfolding and subsequent localization to aggresomes.