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Korean Society of Electron Microscopy
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Volume 18, Issue 2 - Dec 1988
Volume 18, Issue 1 - May 1988
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Ultrastructural and Histochemical Studies on the Epithelial Cells and Mucus-producing Cells of Korean Slug(Limax flavus L.)
Chang, Nam-Sub ;
Applied Microscopy, volume 18, issue 2, 1988, Pages 1~20
The species of the slug used in the experiment is Limax flavus L. For identifying the chemical characteristics of the epidermis, granules and mucus-producing cell of this animal is examined with methylene blue-basic fuchsin double stain and PAS-alcian blue reagent. For the ultrastructural research of the epidermal free surface, the epitheial cell and the parenchymal cell are used with scanning electron microscope and transmission elec-tron microscope respectively. I . Epidermal tissue The epidermal tissue of the slug is observed being divided into the dorsal and the ventral side(toot pad) respectively. 1) Dorsal epidermal tissue The dorsal epidermis of the slug is constituted with the simple columnar epithelium and the microvilli are compacted on the epidermal free surface. Two different types of the secretory granules of the neutral and the acid mucus are observed between the epithelial cells, and the neutral mucous granules are highest electron-dense but the acid mucous granules are observed to be electron-lucent. 2) Foot epidermal tissue The Foot epidermis is formed with the taller simple columnar epithelium than the dorsal epidermis and these cells have both a large number of the microvilli and a few number of the large villi. The secretory granules of three different types, which are acid, neutral and mixed mucous granule of two different types are observed between the epithelial cells. The neutral mucous granules are highest electron dense but the acid mucous granules are observed to be electron-lucent. II . Mucous granule-producing cell and mucus-producing cells Seven different types of the granules-producing cell and the mucus-producing cells are observed between the parenchyma. 1) A-type of acid mucous granule-producing cell The electron-lucent granules are largely occupied in the cytoplasm of these cells and then the granules are surrounded by irregular membrane. These electron-lucent granules exhibit alcianophilia with PAS-alcian blue reaction, so these granules are certified to be acid mucopolysaccharide. 2) B-type of acid mucus-producing cell The nucleus and the cytoplasm of these cells are pushed by the acid mucus of the electron-lucent toward the cell membrane. This mucus has been confirmed to be the acid mucopolysaccharide with PAS-alcian blue reagent. 3) A-type of neutral mucous granule-producing cell These cells contain the electron-dense round granules with approximately
in diameter, which exhibit strongly PAS-positive reaction. These granules are confirmed to be the neutral mucoplysaccharide. 4) B-type of neutral mucous granule-producing cell These cells contain two different types of electron dense granules and electron-lucent granules; The former exhibits to be strongly PAS-positive and the latter to have alcianophilia reaction respectively. 5) C-type of neutral mucus-producing cell These cells are similar to the shape and the size of the B-type of mucus-producing cell but these two different types of cells are stained with reversing properties to each other. The mucus of the C-type cell that electron-lucent is largely occupied in the cytoplasm that exhibits strongly PAS-positive reaction. 6) D-type of neutral mucous granule-producing cell These cells contain round granules about
in size which are observed to be medium electron-dense granules and those granules are stained brightly red with PAS-weak positive reaction. The granules are certified to be neutral mucopolysaccharide. 7) E-type of neutral mucous granule-producing cell These cells are similar to the shape and the size of the D-type of neutral mucous granule-producing cell. These cells contain a large number of granules with about
in diameter showing electron-lucent and then granules are seen to be PAS-weak positive reaction. III. Parenchyma The clear cell and dark cell are found in the parenchyma of the Limax flavus L. 1) Clear cell These cells are round formed and the nucleus of the cells are larger than cytoplasm. These cells which have the electron-lucent cytosol possess poorly developed organelles. 2) Dark cell These cells are found to be dark cells due to high electron-density, which exhibit strongly methylene-blue reaction from double stain of methylene blue-basic fuchsin.
Effect of Chlorambucil on the Ultrastructure of Leydig Cell in Male Mouse
Kim, Douk-Hoon ; Park, Won-Hak ;
Applied Microscopy, volume 18, issue 2, 1988, Pages 21~33
The purpose of this study was to investigate the effect of chlorambucil on the mouse Leydig cell by electron microscopy. Chlorambucil suspended in the 0.5N sodium bicarbonate(pH 8.0) was injected I.P.(intraperitoneal) at a dosage of level 20mg/kg for 1 weeks, 3 weeks and 5 weeks, respectively. The results obtained from this experiment are as follows: 1. One week after the administration of chlorambucil, there was an increase in heterochromatin, swelling and cristae disruption in some mitochondria, mild vacuolation between cells and the occurrence of membrane bound inclusions in some nuclei. 2. After 3 weeks, smooth endoplasmic reticulum dilations, cytoplasmic vacuolation, mitochondrial swelling, inner mitochondrial cristae disruption, membranous whorls, and intranuclear inclusions were observed in the treated cells. 3. After 5 weeks of treatment, most mitochondria were swollen and their membranes were severely disrupted. Further, smooth endoplasmic reticulum dilations and vacuolation of the cytoplasm were apparent in the treated Leydig cells. In addition numerous membranous whorls and intranuclear inclusion bodies were present. The nuclei displayed invaginatons of the nuclear membrane and large clumps of heterochromatin. From these results it is concluded the longer the duration of chlorambucil administration, the greater the degeneration of the nucleus and cytoplasmic organelles.
Changes in the Number of Exocrine Granules in Mouse Pancreatic Acinar Cells Induced by Acetylcholine and MNNG in vitro
Cho, Eng-Haeng ; Choe, Rim-Soon ;
Applied Microscopy, volume 18, issue 2, 1988, Pages 34~46
The stimulation-secretion coupling in the pancreatic acinar cell have been studied by electron microscope. Morphological changes in the cells exhibited the cellular response induced by acetylcholine and MNNG. MNNG, a guanylate cyclase activator, induced the formation of numerous secretory granules in a period after the agent administration. This result suggest that guanylate cyclase potentiated the early sustained response in pancreatic acinar cells stimulated by acetylcholine. Cycloheximide and dibucaine reduced the secretory granules in number during sustained period. In pancreatic acinar cells, the secretion granules were considered to be directly packaged from cisternal space of endoplasmic reticulum.
Structure of Seed Storage Protein and Starch Grains in the Endosperm of Rice Seeds
Park, Kyoung-Soo ; Rho, Young-Bok ; Kaufman, Peter B. ;
Applied Microscopy, volume 18, issue 2, 1988, Pages 47~58
An Immunohistochemical ana Ultrastructural Studies on the Gut Endocrine Cells in the Hedgehog, Erinaceus koreanus
Lee, Jae-Hyun ;
Applied Microscopy, volume 18, issue 2, 1988, Pages 59~76
In order to know the distribution, relative frequencies, types and morphology, endocrine cells in the intestinal tract of the hedgehog(Erinaceus koreanus) were studied by light microscopy, immunohistochemistry and electron microscopy. The results obtained are summarized as follows: 1. Two kinds of endocrine cells were demonstrated with two specific staining methods. Argyrophil cells(reactive cells for Grimelius method) were found most frequently in the intestinal region, and were infrequent in the rectum, whereas argentaffin cells (reactive cells for Masson-Hamperl method) were found most frequently in the rectum and in the other legions were infrequent. These reacting cells were mainly found in the intestinal glands, whereas a small number in the mucosa. 2. Twelve kinds of endocrine cells, gastrin(Gas)-, somatostatin (Som)-, serotonin(5-HT)-, glucagon(Glu)-, bovine pancreatic polypeptide(Bpp)-, cholecystokinin(Cck)-, secretin(Sec)-, motilin(Mot)-, glicentin(Gli)-, gastric inhibitor polypeptide(GIp)-, substance P(Sp)-, and neurotensin(Neu)-immunoreactive cells, were identified by immunohistochemical method. Gas-, Som-, 5-HT-, Glu-, Cck-, Sec-, Mot-, Gli-, Sp-, and Neu-reactive cells were observed in the duodenum, and among these Gas- and 5-HT-reactive cells were moderately found while the others were infrequent. In the jejunoileum Gas-, Som-, 5-HT-, Glu-, Cck-, Mot-, Gli-, GIp-, SP-, and Neu-reactive cells were found, and among these 5-HT- and GIp-reactive cells were moderately found while the others were infrequent. In the colon Sec-reactive cell was not detected. 5-HT-reactive cells were found most frequently and the others were infrequent in this region. 5-HT-, Bpp-, GIi- and Neu-reactive cells were found in the rectum. Among these 5-HT-reactive cells were found most frequently. 3. Electron microscopically, five types of endocrine cells, EC, ECL, D, G, A-like cell, were identified in the intestinal region. EC and ECL cells in the duodenum, EC, D and G cells in the jejuno-ileum, EC and A-like cells in the colon and EC cell in the rectum were observed respectively.
Ultrastructure of the Ampullate Gland in the Orb Web Spider, Nephila clavata L. Koch I. Excretory Duct of the Large Ampullate Gland
Moon, Myung-Jin ; Kim, Chang-Shik ; Kim, Woo-Kap ;
Applied Microscopy, volume 18, issue 2, 1988, Pages 77~90
The ultrastructure of the excretory duct of the large ampullate gland in the orb web spider, Nephila clavain L. Koch are studied with light and electron microscopes. The excretory ducts of the large ampullate glands connected with the large spinning tubes(spigots) on the anterior spinnerets are basically composed of three superposed types of the layers which are inner cuticles, monolayered epithelial cells and peripheral connective cells. According to the morphological characteristics of the cuticles and internal textures of the epithelial cells, the long excretory ducts are subdivided into three(distal, middle and proximal) portions. Especially, at the distal portion of the ducts near the spinning tubes, the electron lucent subcuticles which had the functions of water removal and orientation of silk fibers are well distributed, whereas at the middle and proximal portions these layers disappeared and instead of these, endocuticles are developed. The endocuticle contains two types of bands, which are electron dense and electron lucent. And along the length of the cuticular stem in the excretory duct, these two alternating bands are twisted spirally. In the cytoplasm of the columnar epithelial cells of the distal portion, rough endoplasmic reticula and Golgj complexes, related to the production of the cuticular materials are well developed. Between the adjacent epithelial cells, specialized septate junctions and desmosomes are formed along the plasma membranes. At the proximal portion of the duct, densely accumulated secretory materials appeared, and these are released to the inner canal by the apocrine secretion.
Ultrastructure of the Ampullate Gland in the Orb Web Spider, Nephila clavata L. Koch II. Sac and Tail Portion of the Large Ampullate Gland
Moon, Myung-Jin ; Kim, Chang-Shik ; Kim, Woo-Kap ;
Applied Microscopy, volume 18, issue 2, 1988, Pages 91~101
The ultrastructure of the sac and tail portion of the large ampullate gland and production of the silk materials in the orb web spider, Nephila clavata L. Koch, are studied with electron microscope. Large ampullate glands, the largest glands among the seven kinds of silk glands in this species, are composed of three parts which are the excretory duct, the storage sac and the convoluted tail. The wall of the sac is composed of a single layer of columnar epithelial cells. In the cytoplasm of these cells several kinds of the secretory granules, which are commonly associated with the rough endoplasmic reticula and had characteristic crystalloid, are seen. According to the morphology and internal textures of these granules, the sac portion is subdivided into proximal(connected with the convoluted tail) and distal(connected with excretory duct) portion. Between these two portions, the proximal portion is longer than the distal by four times. Silk materials, being synthesized to the shape of secretory granules, within the glandular epithelial cell of the tail portion are released to the inner cavity by the mechanism of the eccrine secretion. These secretory granules are originated from the rough endoplasmic reticula of the glandular epithelial cells, whereas no Golgi complexes has been found in any of the cells which have been examined.
In vitro Uptake and Accumulation of Purified Storage Proteins into Fat Body Cells from Huphantria cunea Drury
Lee, Bong-Hee ; Kim, Kwan-Seon ; Moon, Myung-Jin ; Kim, Woo-Kap ;
Applied Microscopy, volume 18, issue 2, 1988, Pages 102~118
This study was carried out to examine in vitro first whether the storage proteins, which the fat bodies of last larvae from Hyphantria cunea secrete into haemolymph, can be uptaked by the fat body cells of prepupa and then how the uptaked storage proteins can be accumulated in the fat body cells, if uptaken. The fat bodies which had been isolated from last instar larvae were cultured in 1 ml of Grace's insect medium containing
-leucine (5.0 mCi/mol, Dupont) at
for 6 hrs. After the homogenates of the cultured fat bodies were centrifuged at 10,000 rpm for 10 minutes, the proteins included in the supernatant were separated by polyacrylamide gel electrophoreses (non-SDS, 6%). The next treatment of the electrophoresed gel was followed by rinsing. A storage protein band of several bands in the rinsed gel was sliced off. With elution of sliced storage protein bands in Tris-glycine buffer, the purification of radioactive storage proteins from fat bodies was finished. After the purified radioactive storage proteins were added in Grace's insect midis containing fat bodies of the prepupae, they were cultured for the randomly following minutes given as 3, 5, 7, 10, 15, 20 and 30 and for the randomly following hours given as 1, 2, 3 and 4 respectively. The double fixations of the cultured fat bodies in aldehyde and
, were followed by preparation of ultrathin sections from Epon-Araldite blocks through dehydration and embedding. The electron microscope autoradiographic treatment of all prepared sections were performed by the dipping method (Kim et al., 1987). The finally prepared specimens were examined with electron microscope. The fat body cells of the prepupa could be found to uptake the storage preteins of the last instar larvae, which were included in the culture medium, mostly by formation of coated vesicles. The in vitro uptake of the storage proteins actively occurred by 30 minutes after the addition of purified storage proteins in the culture medium. After culture for 7 minutes with the storage proteins, the uptaked radioactive storage proteins labelled a number of lysosomal granules. After culture for 20 minutes with the storage proteins, the radioactive storage proteins were finally incorporated and accumulated in lipid droplets and protein granules. The frequency in the fat body cell of radiolabelled lipid droplets occurs approximately 60%, while the frequency, in which the radiolabelled protein granules occurs in a fat body cell, is approximately 40%.
A Cytochemical Study on the Acid Phosphatase of Neurons of Developing Chick Embryo Brain
Koh, Ki-Seok ; Shin, Chu-Og ; Yoo, Chang-Kyu ; Choe, Rim-Soon ;
Applied Microscopy, volume 18, issue 2, 1988, Pages 119~131
The purpose of this study was to investigate the differentiation and degeneration of neurons in developing chick embryo. The activity of acid phosphatase(ACP) was measured and cytochemical study of ACP and ultrastructural changes were observed in prosencephalon, mesencephalon and rhombencephalon from day 4 to day 19 of incubation. As a result, the activity of ACP of all brain region was tend to increase from day 4 to day 19. On day 13, activities of ACP of mesencephalon and rhombencephalon were increased greatly and activity of ACP was decreased each region on day 17. On electron microscopic examination, the reaction product of ACP were localized at GERL complex, lysosome, Golgi body and vacuoles of neurons. Morphologically, disrupted nuclear envelope, mitochondrial destruction, vacuolization and ribosomal crystalization were observed.
Ultrastructural Study on the Radulae of Bithyniid Snails (Mollusca: Prosobranehia)
Kim, Jae-Jin ; Choe, Rim-Soon ; Lim, Seung-Sub ; Hahn, Kyu-Woong ; Claus, Meier Brook ;
Applied Microscopy, volume 18, issue 2, 1988, Pages 132~140
The radulae of six species of bithyniid snails, Bithynia striatula, B. tentaculate, B. siamensis, B. leachi, Gabbia misella and G. australis, were observed by SEM. The radular formula of all the bithyniids studied was 2 : 1 : 1 : 1 : 2. The all teeth were multicuspid and the central teeth had numerous basal denticles. The mesocones of the central and lateral teeth were spade or arrow-head shape. The mesocones of the central teeth and the lateral teeth of B. leachi, G. misella and G. australis were slightly larger than the other cusps in contrast of those of the other species. Number of cusp was varied by the specimen and the species.
Studies on the Developmental Processes of the Protein Body in the Ginseng(Panax ginseng C.A. Meyer) Endosperm Cell
Yu, Seong-Cheol ; Jeong, Byung-Kap ; Kim, Woo-Kap ;
Applied Microscopy, volume 18, issue 2, 1988, Pages 141~152
The developmental processes of the protein body are studied on endosperm cells of Panax ginseng during seed maturation periods. The spherosome, mitochondria, rough endoplasmic reticulum, and ribosome are observed and then are gradually increased in early endosperm cells. Protein body developed from vesicles produced by the rough endoplasmic reticulum and was formed at the enlarged ends of rough endoplasmic reticulum. Also, vacuole-like protein body was observed in associated with rough endoplasmic reticulum. Golgi complex is observed in associated with vacuole and its vesicles containing proteinaceous granules moved and accumulated to the vacuole. Proteinaceous granules are deposited in the spherical or oval shaped vacuole and gradually, vacuole is surrounded by the multi-membranous structure. Rough endoplasmic reticulum, ribosome, Golgi complex, and vacuole are observed in associated with protein body formation.
Fine Structural Study on the Salivary Glands in the Pine Moth, Dendrolimus spectabilis Butler at the Last Larval Period
Baek, Seung-Bum ; Moon, Myung-Jin ; Kim, Woo-Kap ;
Applied Microscopy, volume 18, issue 2, 1988, Pages 153~166
The fine structure of the salivary glands of the pine moth, Dendrolimus spectabilis Butler, at the last larval period is observed using light and electron microscopes. The moths have single paired tubular salivary glands which openings are connected to the oral cavity through the upper jaw. By the external morphology and its functions, the glands are subdivided into three regions which are anterior reabsorptive region, middle storage region and posterior secretory region. Along the inner canal of the salivary gland two columns of the large glandular cells are connected each other and oriented to ring-like forms. By this cellular orientation, the glands have long and large tubular structure. From anterior to posterior region large nuclei of the glands are ramified like twigs of the tree, and in the cytoplasm of the cell numerous mitochondria and vacuoles are seen. Moreover, basal plasma membranes of the gland cells are heavily infolded. The anterior region of the glands keeps several characteristics related to the reabsorption of the material from the inner cavity to the glandular cells whereas, main salivary material is synthesized and secreted through the long and convoluted posterior region. The apical plasma membranes of the cells are the most heavily invaginated at the posterior regoin, but trachea and tracheoles are distributed only at the middle and posterior regions. In the cytoplasm of the middle region Golgi complexes appeared at the vicinity of the vesicles, and at the posterior region of the salivary glands multivesicular bodies are also observed.
Electron Microscopic Study on Extrachromosomal DNA from Splenocytes and Erythrocytes of Carassius carassius L.
Im, Sook-Ja ; Kim, Woo-Kap ;
Applied Microscopy, volume 18, issue 2, 1988, Pages 167~176
Extrachromosomal circular DNA complexes from erythrocytes and splenocytes isolated from Carassius carassius were examined by mica-press-absorption method. The method was described that released small polydisperse circular DNA molecules in situ from the erythrocytes and the splenocytes and that allows selective observation of the small circular DNA complexes bound to cellular components. The released polydisperse circular DNA complexes were absorbed preferentially on mica in a divalent cation-free medium then processed for electron microscopy. Small circular DNAs showed a heterogeneous size distribution of
with a mean contour length of
for the circulating erythrocytes and that of
with a mean contour of length
for the splencytes. Cells contained
copies obtained from the erythrocytes and the splenocytes, repectively. Possible biological functional implications for size distribution of extrachromosomal circular DNAs are discussed.
Ultrastructural Changes in the Neuropil of the Anterior Thalamic Nucleus following the Lesion in the Mamillary Body
Lee, Byoung-Ho ; Ko, Jeong-Sik ; Ahn, E-Tay ; Yang, Nam-Gil ;
Applied Microscopy, volume 18, issue 2, 1988, Pages 177~186
Degeneration of the axon terminals of mamillo-thalamic tract following the electrical coagulation of mamillary body is well known. In this study, the author investigated the ultrastructural alterations of neuropil components, initiated by terminal degenerations. Rats weighing approximately 250 gm were fixed on the stereotaxic instrument(David Kopf Inc., Heavy duty model), and NE 300 active electrode(Rhodes Med. Instr. Inc.) was introduced to the mamillary position of anterior 3.8 mm, lateral 0.5 mm, height 3.8 mm and lateral angle of
according to De Groot's Atlas. Electric current of 20 mA was applied during 1 minute between active and inactive electrodes with Radio Frequency Lesion Generator(RFG 4, Radionics Inc.). Two hours, 2 days, 1 week and 2 weeks following the electrical coagulation of mamillary body, ipsilateral anterior thalamic nucleus was fixed in 1% glutaraldehyde-l% paraformaldehyde and 2% osmium tetroxide, embedded in Araldite mixture, cutted with LKB ultra tome V, stained with uranyl acetate-lead citrate and observed with JEOL 100 CX electron microscope. Observed results were as follows; 1. Degenerated mamillo-thalamic synapses were observed to form asymmetric axospinous or axo-dendritic types. 2. Terminal degeneration was not easily discernible at 2 hours interval after mamillary lesion, but following 2 days the terminal degeneration was apparent. 3. Postsynaptic spines, dendrites and even their cell bodies show edematic changes caused by the degeneration of postsynaptic counterpart. 4. Astrocytic territories, including perivascular processes forming glial limitans of blood-brain barrier, exhibit remarkable expansion. 5. Oligoglia and astroglia are actively engaged in the removal of degenerated elements. 6. Active forms of microglia were increased. 7. The observed results may represent typical ultrastructural alteration pattern within neuropil following the degeneration of certain input axon terminals.
Acute and Subacute Effect of Lead acetate on Enzyme Activities and Ultrastructure in Mouse Diencephalone
Lee, Jung-Hee ; Yoo, Chang-Kyu ; Choe, Rim-Soon ;
Applied Microscopy, volume 18, issue 2, 1988, Pages 187~204
The present experiment was performed to investigate the acute and subacute effect of lead acetate on ultrastructural and biochemical changes in mouse diencephalon. In acute case, mouse were peritoneally injected with lead acetate at a dose of 0.26 mmole/kg body weight, and after treatment, mouse were sacrificed at time intervals of 12, 24, 48, and 96 hours. In subacute case, mouse were injected at doses of 0.07 mmoie/kg B. W. and 0.13 mmole/kg B.W. once at two days, and after treatment, mouse wee sacrificed at 1 week, 2 weeks, and 3 weeks. It was observed that after acute treatment, changes composed of increased monoamine oxidase activity,
ATPase activity, decreased
-APTase activity, wrinkled myelin, swollen Golgi apparatus and more dense synaptic vesicle in nerve terminal. After subacute treatment, decreased monoamine oxidase activity, increased
ATPase, lose of myelin, uneven mitochondrial distribution, synaptic vesicular density and edema, but at a higher dose the effect was more severe. Therefore, lead acetate caused abnormal change of diencephalon, and at a subacute, it appears metal accumulative toxicity.
Observation of Histochemical Ultrastructure in Regenerating Rat Liver
Choi, Chee-Yong ; Sohn, Seong-Hyang ; Yoo, Chang-Kyu ; Choe, Rim-Soon ;
Applied Microscopy, volume 18, issue 2, 1988, Pages 205~217
An ultrastructural study of hepatocyte proliferation in the regenerating rat liver has been made by means of the partial hepatectomy. And electron microscopic histochemistry of hepatocyte in the regenerating rat liver is studied through alkaline phosphatase reaction. The results are as follows: 1. When the regeneration of rat liver is induced by the partial hepatectomy, the prominent ultrastructural characteristics of hepatocyte are changes of the distribution of chromatin in nucleus, increase of the number of mitochondria and decrease of the size of them, development of rough endoplasmic reticulum, and transient decrease of glycogen granules in cytoplasm. 2. Alkaline phosphatase reaction products are appeared in the nucleus or rough endoplasmic reticulum of hepatocyte during the initial regeneration of liver as 24, 48 and 72 hour groups after partial hepatectomy. And these positive reaction are mainly increased in cytoplasm and plasma membrane of hepatocytes during 1, 2 and 3 week groups after partial hepatectomy. As 4 weeks passed after partial hepatectomy, these positive reaction is located in the sinusoidal epithelial cells or erythrocytes. With above results, we concluded that alkaline phosphatase was synthesized in the rough endoplasmic reticulum bounded ribosomes of regenerating hepatocyte, was transported to the plasma membrane of them, and then was transported in blood by the way sinusoidel epithelial cells.