• Applied Microscopy
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• v.31 no.2
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• pp.157-165
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• 2001

Sertoli cells in the normal adult testis are nondividing cells, which are relatively inconspicuous on cross section of the seminiferous tubule and comprise about 10% to 15% of the tubular cellular elements. Ultrastructurally, Sertoli cells have characteristic nucleoli, plasma membrane, and cytoplasmic components. The plasma membrane has two types of intercellular junctions which are developed at puberty: junctions between adjacent Sertoli cells and Sertoli cell-germ ceil junction. However, the ultrastructural findings of Sertoli cells in human fetus is not fully elucidate yet. In the present study, human fetal testes ($14\sim27$ weeks) obtained from artificially induced abortions legally without gross malformation were studied using transmission electron microscopy to make clear the differentiation process of Sertoli cells in human. In human fetal testes from 14 weeks to 27 weeks, the cell junctions of Sertoli-germ cells and Sertoli-Sertoli cells are desmosome like structure and not tight junction or desmosome. The Overall intracytoplasmic organelles of Sertoli cells are relatively sparse. The mitochondrias are relatively abundant but no developed cristae. And the rough endoplasmic reticuli are abundant and smooth endoplasmic reticuli are sparse. The amount of lipid droplets are regularly observed in human fetal Sertoli cells. No microfilaments or Charcot-Bottcher's crystalloids are present. From the results, Sertoli cells in human fetal testes are somewhat different ultrastructural findings with puberty or adult. However, to make clear the differentiation process of Sertoli cells in human, further study for 28 weeks to puberty is required.

• Applied Microscopy
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• v.24 no.2
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• pp.48-62
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• 1994

The acute irradiation effect on rat Purkinje cell was carried out. Anesthetized rats, weighing 200-250g each, were exposed their heads to the linear accelerator (ML-4MV) with the doses of 3,000 rads or 6,000 rads respectively. Irradiated rats were sacrificed by perfusion fixation under anesthesia, six hours, two days and six days following the irradiations. Rats were perfused with the fixative of 1% glutaraldehyde-1% paraformaldehyde solution (pH 7.4). Small pieces of cerebellar cortices were taken out. Tissue blocks were washed out, and were refixed in the 2% osmium tetroxide solution. After dehydration, tissues were embedded in the araldite mixture. Ultrathin sections stained with uranyl acetate-lead citrate solution, were examined with an electron microscope. The results observed were as follow; 1. Many dark Purkinje cells exhibited most severe cellular alterations on 6 hours. But after the 2 or 6 days, the cells exhibited only some alterations of cytoplasmic organelles. 2. Many granular and agranular endoplasmic reticula exhibited the fusion of cisterns. These reticular alterations were most severe on 6 hours following irradiation. But the alterations were hardly found on 6 days. 3. In the Golgi region, alterations including the adhesion of lamelliform cisterns, enlarged saccules, and increased number of vesicles, etc, were seen on 6 hours. But the Golgi complexes were almost recovered on 6 days. 4. Lysosomes were abundant on 6 hours or 2 days, but some residual bodies were found on 6 days. 5. Mitochondrial changes were also most severe at on hours, and they were recovered thereafter. From the results, it was concluded that the cerebellar Purkinje cells reacted to the high doses of irradiation by hyperactive protein synthesis, autolytic activities and energy metabolism. The reaction was most active in the early stage. It implies that motor-control function of Purkinje cells are severely disturbed in the early stage of irradiation.

• The Korean Journal of Zoology
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• v.31 no.1
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• pp.35-48
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• 1988

This study was made to investigate the effect of testosterone and cyclic-AMP (cAMP) on rat epididymis. Peritoneai injections of testosterone and cAMP to rats were earned out The activities of acid phosphatase (ACP), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) were measured and ultrastructural changes of luminal epithelial cells were observed. As a result, the activity of ACP was significanily decreased on the third day, that of ALP on the fifth day and that of LDH on the seventh day respectively in castrated group. In addition, the activities of ACP and ALP were significanily increased when treated with testosterone for 5 days, that of LDH when treated with testosterone for 7 days. In case of cAMP and cAMP - theophylline injection, the activities of ACP and LDH were increased but the range of increase was of no significance. However a significant increase in the activity of ALP was seen on both cases. On electron microscopic examination, gradually deformed Golgi complex, destructed mitochondria and disrupted stereociha were observed in castrated group. In case of testosterone injection, disrupted Golgi complex, mitochondria and stereocilia showed recovery. When cAMP and cAMP-theophylline were injected as an alternative, various cytoplasmic organelles as well as Golgi com- plex were recovered but stereocilia remained unrecovered.

• Tuberculosis and Respiratory Diseases
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• v.69 no.2
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• pp.81-94
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• 2010

Background: Autophagy is an important adaptive mechanism in normal development and in response to changing environmental stimuli in cancer. Previous papers have reported that different types of cancer underwent autophagy to obtain amino acids as energy source of dying cells in nutrient-deprived conditions. However, whether or not autophagy in the process of lung cancer causes death or survival is controversial. Therefore in this study, we investigated whether nutrient deprivation induces autophagy in human H460 lung cancer cells. Methods: H460, lung cancer cells were incubated in RPMI 1640 medium, and the starved media, which are BME and RPMI media without serum, including 2-deoxyl-D-glucose according to time dependence. To evaluate the viability and find out the mechanism of cell death under nutrient-deprived conditions, the MTT assay and flow cytometry were done and analyzed the apoptotic and autophagic related proteins. It is also measured the development of acidic vascular organelles by acridine orange. Results: The nutrient-deprived cancer cell is relatively sensitive to cell death rather than normal nutrition. Massive cytoplasmic vacuolization was seen under nutrient-deprived conditions. Autophagic vacuoles were visible at approximately 12 h and as time ran out, vacuoles became larger and denser with the increasing number of vacuoles. In addition, the proportion of acridine orange stain-positive cells increased according to time dependence. Localization of GFP-LC3 in cytoplasm and expression of LC-3II and Beclin 1 were increased according to time dependence on nutrient-deprived cells. Conclusion: Nutrient deprivation induces cell death through autophagy in H460 lung cancer cells.

• Applied Microscopy
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• v.29 no.1
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• pp.105-124
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• 1999

Histological and ultrastructural differentiations of the neuroepithelial cells in the mouse embryo during neurulation were observed. The neural plates and grooves consisted of pseudostratified columnar epithelium in the embryonic day (ED) 8 embryo were developed. In the ED 9 embryo, the neural tube was developed in all body length of embryo except both the cephalic and caudal ends. Secondary neurulation was shown at the tail bud of the ED 10 embryo. In the ED 8 embryo, the primitive streak was shown in the posterior end of the embryonic disc. The neuroepithelium, notochord and mesenchyme were well differentiated in the cephalic and cervical portions. In the ED 9 and 10 embryos, the roof plates of neural tubes were constituted of the closing of the surface ectodermal cells in the hindbrain and the neuroepithelial cells in the spinal cord. The floor plate of neural tube were consisted of the low pseudostratified columnar epithelium. The spinal motor nerve fibers were initially differentiated in the ED 10 embryo. According to the electron density of the cell and the differentiation of tell organelles, the neuroepithelial cells in the ED 9 and 10 embryos were classified into three types: dark, intermediate and light types. All types in the ED 9 embryo were observed but the dark cell in the ED 10 embryo was not done. The free ribosomes and polysomes in all neuroepithelial cells were developed. The RER and lipid droplets in the dark cell and the Golgi complex in the intermediate and light cells were observed. Many microfilaments in the cytoplasmic processes of intermediate cell and the microfilaments and microtubules in the light cell processes were observed to be well differentiated.

• Parasites, Hosts and Diseases
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• v.36 no.1
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• pp.7-14
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• 1998

In order to understand the action mechanism of polyhexamethylene biguanide (PHMB) to the cyst of Accnthcnloebc on the morphological basis, the cysts of four corneal isolates of Acanthanoebc were treated with minimal cysticidal concentration (MCC) of PHMB and their ultrastructural changes were examined by transmission electron microscopy. The most striking change of cysts treated with PHMB compared with normal cysts was the shrinkage of intracystic amoebae, which resulted in the separation of the plasma membrane of intracystic amoeba from endocystic wall. Subplasmalemmal lipid droplets became irregularly shaped . In severely damaged cysts, cytoplasm was aggregated and organelles were severely deformed. Cytoplasmic materials were leaked out through the damaged plasma membrane. Most cysts showed aggregation of nuclear chromatin material. Number of mitochondrial cristae was also reduced. Ecto- and endo-cystic walls were relatively well tolerated. Findings in the present study revealed that PHMB affected mainly on plasma membrane, but lesser on organellar membrane of intracystic amoeba. It seemed likely that PHMB might kill cystic forms of Accnthamoebc by similar mechanism in which this environmental biocide can damage the cell wall of Escherichia coli by binding with acidic phospholipids.

• Applied Microscopy
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• v.15 no.2
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• pp.31-68
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• 1985

Authors are trying to unveil the ultrastructural organization of olfactory bulb, which has been summerized under light microscopic level or communicated only in some detail in different view point until now. For the critical point of view, since the phylogenetical approach will give the ultimate value in the correlative study between structural and functional bases (Brodal, 1969), the present study was carried out light and electron microscopic analyses of the structures of the neurons and synaptic organizations in olfactory bulbs from different animals in phylogenetical scale. We selected each one species from five animal classes: the house rabbit(Oryctolagus cuniculus var. domesticus [Gmelin]) from Mammalia, the domestic fowl (Gallus gallus domesticus Brisson) from Aves, the viper (Agkistrodon hylys [G.P. Pallas]) from Reptilia, a frog (Bombiana orientalis Boulenger) from Amphibia and the crussian carp (Carassius carassius [Linne]) from Pisces. For light microscopic study, samples were fixed in 10% formalin and paraffin sections were stained with hematoxylin-eosin. For the electron microscopic study, the tissues were fixed by perfusion through the heart or immersion with 1% paraform-aldehyde-glutaraldehyde mixture (phosphate buffer, pH 7.4), and final tissue block trimmed under dissecting microscope were osmicated (1% OsO4), they were embedded in Araldite or Epon 812, and ultrathin sections were made by LKB-V ultratome following the inspection of semi-thin sections stained with toluidine blue-borax solution. Ultra-thin sections contrasted with uranyl acetate and lead citrate were observed with JEM 100CX electron microscope. We have summerized our morphological analyses as follows: 1. The olfactory bulb of rabbit, viper and frog shows the eight layers of fila olfactoria, glomerular, external granular, external plexiform, mitral cell, internal plexiform, internal granular, medullary but domestic fowl shows the five layers of glomerular, fibrillar, mitral, granular and medullary and the three layers of fibrilla, glomerular and medullary in crussian carp. The sharpness of demarcation between the layers shows deferential tendency according to phylogenetical order. 2. Mitral cells of vertebrate have large triangular or oval shape with spherical nuclei which contain not so much chromatin. The cytoplasm contains numerous cell organelles, of which Nissl's bodies or granular endoplasmic reticula arranged as parallel strands. Development of granular endoplasmic reticula were declined as the phylogentical grade is going lower. 3. Tufted cells of all animal are mostly spindle or polygonal contour and contain oval nuclei which located in periphery of cytoplasm. The nuclei of rabbit, fowl, viper and frog has relatively space chromatin, but a nucleus of crussian carp contain irregularly aggregated chromatin in karyoplasm. Their cytoplasmic volume and cell organelle contents are in between those of mitral cell and granular cell. They contain moderate amount of mitochondria, granular endoplasmic reticula, a few Golgi complex, polysomes, lysosome, etc. 4. Granule of cells of all the vertebrate amimals studied exhibit similar features; cells and their dense nuclei show spherical or oval contour, and they have the thin rim of cytoplasm which contain only a few cell organelles. 5. In rabbit, the soma of mitral cells were in contact with boutons with two types of synaptic vesicles, that is, round and flat vesicles, especially flat vesicles in boutons were showing reciprocal synapses. However, in domestic fowls, vipers, frogs and crussian carps, there were found boutons showing only spherical synaptic vesicles. 6. The boutons containing round synaptic vesicles were made contact with the some of tufted cell of olfactory bulb in the rabbits, fowls, vipers and frogs, but no synaptic boutons were observed in soma of tufted cells in crussian carps. In the frogs, there were observed dendrites were contact with the soma of tufted cells. 7. In the neuropils of plexiform, granular and glomerular layers olfactory bulbs in the vertebrate, the synapses were axo-large dendrites, axo-median and small dendrites, dendrodendritic, and axo-axonal contacts. However, in the neuropil of crussian carps, synapses were observed only in glomerular layer.

• Journal of Life Science
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• v.33 no.7
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• pp.531-537
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• 2023

Intracellular and axonal transport is mediated by microtubule-dependent motor proteins, such as kinesins and cytoplasmic dynein. Kinesin moves along the microtubule to the positive end of the microtubule, while dynein moves to the negative end of the microtubule. Kinesin-1 was first identified as a kinesin superfamily protein (KIF) that functions in the intracellular transport of various cargoes, including organelles, neurotransmitter receptors, and mRNA-protein complexes, through interactions between the carboxyl (C)-terminal domain and the cargo. It interacts with other cargoes, but the adapter/scaffold proteins that mediate between kinesin-1 and the cargo have yet to be fully identified. In this study, a yeast two-hybrid screen was used to identify adapter proteins that interact with the C-terminal region of KIF5A. We found an association between the C-terminal region of KIF5A and the cyclin-dependent kinase 2-associated protein 1 (CDK2AP1), originally identified in malignant hamster oral keratinocytes. CDK2AP1 bound to the C-terminal region of KIF5A and did not interact with KIF3A (the motor of kinesin-2), KIF5B, KIF5C, and kinesin light chain 1 (KLC1). The C-terminal region of CDK2AP1 is essential for its interaction with KIF5A. When co-expressed in HEK-293T cells, CDK2AP1 and kinesin-1 co-immunoprecipitated and co-localized in the cells. These results suggest that the KIF5A-CDK2AP1 interaction serves as an adapter protein connecting kinesin-1 and the cargo when kinesin-1 transports cargo in cells.

• Applied Microscopy
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• v.25 no.4
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• pp.83-103
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• 1995

This experiment was carried out to investigate the effects of telluric acid on the histological and fine structural changes in the rat liver. Fischer 344 rats($150{\sim}200gm$) were used in this study as control and experimental groups. Telluric acid(5 mg/100 gm of body weight) suspensed in olive oil was given intraperitoneally to the animals of the experimental group and only olive oil to those of the control group. At the intervals of 3, 6 and 12 hours, 1, 2, 3, 5, 10, 20, 30 and 60 days after administration, the animals were sacrificed, and livers were obtained from the rats. For light microscopic examination of the liver, sections($5{\mu}m$) were stained with hematoxylineosin(H-E). For electron microscopic examination of the liver, sections were stained with uranyl acetate and lead citrate, finally examined with Zeiss EM 109 electron microscopes. The results obtained were as follows. 1. In the control group, round nucleus. well developed mitochondria, Golgi apparatus, rough endoplasmic reticulum(RER) and numerous glycogen particles were observed in the cytoplasm of the hepatocyte. In the cytoplasmic membranes of the hepatocyte, sinusoidal surface had numerous microvilli and cellular surface is combinated adjacent hepatocyte with desmosomes. The RER cisterns were dilated and zymogen granules were fewer than those of the dark cells. Kupffer cells with irregular nuclear membrane were observed. Fat storing cell and collagenous fiber bundle were observed in the Disse space. 2. Kupffer cell, inflammatory cells in the connective tissue of hepatic triad and lysosome were increased in the 3, 6, and 12 hour experimental group comparing with that of the control group. 3. In the 1 day experimental group, infiltration of inflammatory cells in interlobular connective tissue, dilatation of sinusoidal capillary and increasing of Kupffer cell were observed. Atropic change of hepatocyte and aggregation of glycogen particles in the cytoplasm of hepatocyte were observed. In this group, desmosome near bile canaliculi and collagenous fiber bundle in the Disse space were increased comparing with that of the 12 hours experimental group. In the 2 days experimental group, desmosome, lysosome, peroxisome and collagenous fiber bundle were increased comparing with that of the 1 day experimental group. Furthermore, lamellated bodies were also seen in the cytoplasm of the hepatocyte. 4. In 3 and 5 days experimental groups, transformations of hepatic cell cord and degeneration of the hepatocyte were markedly inclosed comparing with the all experimental groups. And damaged RER and mitochondria. collagenous fiber bundle were also inclosed comparing with that of the 2 days experimental group. Autophagosome and fat storing cells with large lipid droplets were also observed comparing with that of the 2 days experimental group. Tight junction and desmosome between the hepatocytes were separated. These degenerating changes were severe through the all experimental groups. 5. In the 10 and 20 days experimental groups, arrangement of hepatic cell cords and cell organelles of hepatocytes were similar to those of the control group. However, aggregation of glycogen particles, dilatation of sinusoidal capillary and infiltration of inflammatory cells remained. 6. In the 30 days experimental group, the tissue findings were similar to those of the control grout. But lamellated bodies in some hepatocytes and lysosome were remained in the cytoplasms of the Kupffer cells. In the 60 days experimental group, these all changes were recovered as the control group. In conclusion, telluric acid would directly induce the degenerative and necrotic changes on the hepatic tissue. However, these changes were perfectly recoverd in the 60 days experimental group as the control group.

• Journal of Life Science
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• v.33 no.1
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• pp.1-7
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• 2023

Intracellular cargo transport is mediated by molecular motor proteins, such as kinesin and cytoplasmic dynein. Kinesins make up a large subfamily of molecular motors. Kinesin-1 is a plus-end-directed molecular motor protein that moves various cargoes, such as organelles, protein complexes, and mRNAs, along a microtubule track. It consists of the kinesin superfamily protein (KIF) 5A, 5B, and 5C (also called kinesin heavy chains) and kinesin light chains (KLCs). Kinesin-1 interacts with many different binding proteins through its carboxyl (C)-terminal region of KIF5s and KLCs, but their binding proteins have not yet been fully identified. In this study, a yeast two-hybrid assay was used to identify the proteins that interact with the KIF5A specific C-terminal region. The assay revealed an interaction between KIF5A and glutamate-rich 4 (ERICH4). ERICH4 bound to the KIF5A specific the C-terminal region but did not interact with the C-terminal region of KIF5B or KIF3A (a motor protein of kinesin-2). In addition, KIF5A did not interact with another isoform, ERICH1. Glutathione S-transferase (GST) pull-downs showed that KIF5A interacts with GST-ERICH4 and GST-ERICH4-amino (N)-terminal but not with GST-ERICH4-C or GST alone. When co-expressed in HEK-293T cells, ERICH4 co-localized with KIF5A and co-immunoprecipitated with KIF5A and KLC but not KIF3B. Together, our findings suggest that ERICH4 is capable of binding to KIF5A and that it may serve as an adaptor protein that links kinesin-1 with cargo.