• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.73-73
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• 2003

The present study examined the possibility of long term storage, by cryopreservation in liquid nitrogen, of the sperm of Filefish (Thamnaconus septentrionalis), and the changes in motility, survival rate and ultrastructure of the sperm after freezing and thawing. The sperm was collected by stripping and stored on ice until experiments. For selection of the immobilizing solution, diluted artificial seawater (ASW) of 20, 30 and 40% were tested. The sperm motility was significantly inhibited in 30% ASW, and restored entirely after 100% ASW was added again. Two cryoprotectants, dimethyl sulfoxide ($Me_2$SO) and glycerol, were added to 30% ASW to formulate the extenders at the concentrations between 5 to 20% by volume for freezing. The sperm was diluted at the ratio of 1 :6 with the extenders, inserted into 0.5ml plastic straws and frozen at a freezing rate of $50^{\circ}C$/min to $-100^{\circ}C$ after equilibration for 10 min at room temperature, followed by plunging into liquid nitrogen. The straws were thawed in a $30^{\circ}C$ water bath for 15 sec. The highest post-thawed sperm motility and survival rate were obtained with 5% glycerol Afterward, the effect of different freezing rates was examined using 5% glycerol as a cryoprotectant, and the rate of $20^{circ}C/min to $-80^{\circ}C$ showed the best result Some ultrastructural changes of sperm, such as the detachment of plasmatic and nuclear membranes, destruction of mitochondria, were observed after cryopreservation. Morphological normality of the sperm in 5% glycerol frozen at the ratio of 1$0^{\circ}C$/min to $-80^{\circ}C$ was better than that of others.

• Journal of Veterinary Science
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• v.25 no.5
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• pp.61.1-61.13
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• 2024

Importance: Ovine pulmonary adenomatosis (OPA) and maedi-visna disease (MVD) are chronic and progressive infectious diseases in sheep caused by Jaagsiekte sheep retrovirus (JSRV) and maedi-visna virus (MVV), respectively. Objective: To investigate the pathological changes and conduct viral gene analysis of OPA and MVD co-occurrence in Inner Mongolia, China. Methods: Using gross pathology, histopathology, immunohistochemistry, ultrastructural pathology, PCR, and sequence analysis, we investigated the concurrent infection of JSRV and MVV in 319 Dorper rams slaughtered in a private slaughterhouse in Inner Mongolia, in 2022. Results: Of the 319 rams included, 3 showed concurrent JSRV and MVV infection. Gross lung pathology showed diffuse enlargement, consolidation, and greyish-white miliary nodules on the lung surface; the trachea was filled with a white foamy fluid; hilar and mediastinal lymph nodes were significantly enlarged. Histopathology results revealed typical OPA and MVD lesions in the lung tissue. Immunohistochemical results were positive for JSRV envelope protein (Env) in the tumor cells and MVV CA in alveolar macrophages. Transmission electron microscopy showed several virions and autophagosomes in the lung tissue, severely damaged mitochondria, and the induced mitophagy. Nucleotide sequences obtained for JSRV env and MVV gag showed the highest homology with the Inner Mongolian strains of JSRV env (JQ837489) and MVV gag (MW248464). Conclusions and Relevance: Our study confirmed that OPA and MVD co-occurrence and identified the pathological changes in Inner Mongolia, China, thereby providing references for the identification of concurrent JSRV and MVV infections.

• The Korean Journal of Zoology
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• v.25 no.1
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• pp.9-28
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• 1982

Cyclical changes in the fine structures of the surface epithelial, stroma and glandular cells of guinea pig endometrium during the estrous cycle were studied by transmission and scanning electron microscopy. Cytochemical studies were made in order to investigate the ultrastructural localization of the acid phosphatase, alkaline phosphatase and ATPase in these cells. The results obtained are as follows: 1. The endometrial surface epithelium was pseudostratified columnar during estrus and meterstrus, and simple columnar during proestrus and diestrus. The characteristic features observed in these cells include increased nucleocytoplasmic ratio at proestrus, elongated shapes of both the nucleus and the entire cell, increased volume of the cytoplasm and cytoplasmic bulding into the lumen during estrus, and smaller surface epithelial cells during metestrus. 2. In the cytoplasm of surface epithelial cells, the numbers of mitochondria and free ribosomes were increased, and rough endoplasmic reticulum and Golgi complex appeared during estrus, and the degenerated cells, lipid droplets, multilamellated bodies and lysosomes appeared during diestrus. 3. During estrus, scanning electron microscopic observations of endometrial surface showed a regular arrangement with polygonal outlines of epithelial cells, distinct intercellular border, and bulged surface into the lumen, whereas flat surface and indistinct cell border were characteristic during meterstrus and diestrus. 4. Microvilli which aligned on the surface were longer and most abundant during estrus while short and aparse during other phases. 5. Cytochemical studies indicated that during metestrus acid phosphatase activities were localized in the microvilli and vacuoles, and alkaline phosphatase activities were significant around luminal surface and lateral cell membrane in the surface epithelial cells. ATPase activities were present on the microvilli and cell membrane during proestrus and estrus.

• Development and Reproduction
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• v.12 no.1
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• pp.41-49
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• 2008

Ultrastructural changes occurring during the course of development and degeneration of oocytes in female Ruditapes philippinarum (Adams & Reeve, 1850) are described for clams collected from Gomso Bay, Korea. During the early stages of oogenesis, desomosome-like gap junctions localized between the early vitellogenic oocyte and the follicle cells. Vitellogenesis occurs through a process of autosynthesis, involving the combined activity of the Golgi complex, mitochondria and rough endoplasmic reticulum, and heterosynthesis in which extraovarian precursors are incorporated into oocytes by endocytotic activity, involving the basal region of the early vitellogenic oocytes prior to the formation of the vitelline envelope. The follicle cells appear to play an integral role in vitellogenesis and oocyte degeneration: phagocytosis and intracellular digestion of products originating from oocyte degeneration. These functions can permit a transfer of yolk precursors necessary to vitellogenesis, and they can accumulate nutrients in the cytoplasm, as glycogen and lipids, which can be employed by the vitellogenic oocyte. During the period of oocyte degeneration, follicle cells may have lysosomal system for breakdown, and resorb various phagosomes in the cytoplasm for nutrient storage. But follicle cells probably are not the major source of yolk precursors in vitellogenesis.

• The Korean Journal of Malacology
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• v.25 no.2
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• pp.145-151
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• 2009

The ultrastructural changes in germ cells during oogenesis of the melania snail, Semisulcospira libertina libertina have been investigated by light and electron microscopy. The ovary is located on the surface of the hepatopancreas in the spiral posterior region. The ovary exhibited greenish color in the gonadal mature season. The ovary was composed of a number of oogenic follicles. Oogenesis was divided into five stages with histological features: (1) oogonia, (2) previtellogenic, (3) initial vitellogenic, (4) active vitellogenic, and (5) mature stages. Oogonia were oval in shape, $4-6\;{\mu}m$ in diameter, and had a large nucleus. Previtellogenic oocytes were about $20\;{\mu}m$ in diameter and the cytoplasm reacted with hematoxylin in H-E satin. Initial vitellogenic stage, oocytes were $60-80\;{\mu}m$ in diameter, and small yolk granules of low electron density are scattered in the cytoplasm. Oocytes in the initial vitellogenic stage were connected with ovarian follicle by egg stalk. Active vitellogenic oocyte were $100-120\;{\mu}m$ in diameter. Electron density, size and quantity of yolk granules that are distributed in the cytoplasm have increased from the previous stage. Result of TEM observations, the oocyte contains well-developed Golgi complex, endoplasmic reticula and tubular mitochondria in the cytoplasm. Cytoplasm of mature oocyte was filled with proteinaceous yolk globules of high electron density. In this stage, the length of microvilli in the egg envelope was approximately $1.1\;{\mu}m$.

• Applied Microscopy
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• v.22 no.2
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• pp.15-29
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• 1992

The gallbladder is known to have the function of the storage and the concentration of the bile produced by the liver. This function is carried out by the removal of water and inorganic electrolytes. Extrahepatic cholestasis or the impairment of excretion of the bile leads to the distension and loss of the function of the gallbladder. The purpose of this study was to examine the ultrastructural characteristics of the normal gallbladder epithelial cells, and their structural changes induced by the ligation of common bile duct of the rabbit. Common bile duct ligation was performed under ether anesthesia. The rabbits were sacrificed on the 1st, 3rd, 5th, 7th and 14th day, respectively after operations. The tissue blocks of the gallbladder were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde prior to fixation in 1% osmium tetroxide, and embedded in the araldite mixture, and observed with JEM 100 CX-II electron microscope. The results were as follows: 1. The normal gallbladder epithelium of adult rabbit demonstrated two cell types, the ordinary epthelial cell and the dark cell. The dark cells have electron dense cytoplasm, and were found much infrequently, whereas ordinary epthelial cells were found quite numerous. 2. The ordinary epthelial cells of normal gallbladder were provided with the regular microvilli at the free surface and the images of pinocytotic activities in the apical cytoplasm, and exhibit highly convoluted lateral surfaces with elaborated microfolds. These figures of the cells suggest that they are resorptive in functional activity. 3. In the early stages (1st, 3rd, 5th day groups) following the ligation, the apical cytoplasm of some cells is protruding from the free surface and lost their microvilli. Numerous mucous granules filled in the apical and supranuclear cytoplasm compactly. 4. In the late stages (7th, 14th day groups) following the ligation, many light cells containing mumerous mucous granules are seen, between the ordinary epthelial cells. Mucous granules are fused each other, and are discharged into the lumen from the apical cytoplasm. The lateral membranes are straight or undulating without any interdigitations. From the above results, it was concluded that in the cholestasis induced by the common bile duct ligation, there is a tendency for the mucosal epithelium of the rabbit gallbladder to have secretory rather than an absorptive function.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• v.22 no.1
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• pp.43-53
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• 1992

The purpose of this study was to investigate the effects of irradiation on the microvascular structure of the submandibular gland in rats. For this study, 110 male rats were singly irradiated with the dose of 10Gy or 20Gy to their neck region by 6MV X-irradiation and sacrificed on the 1st, 3rd, 7th, 14th and 28th day after irradiation. The author observed distribution and structural changes of the microvasculature in rat submandibular glands using a scanning electron microscope by forming vascular resin casting. The author observed ultrastructural changes of the endothelial cells using a transmission electron microscope, and also histologic changes using a light microscope at Hematoxylin and Eosin staining and PAS staining process. The results of the irradiation effects on the microvasculature in rat submandibular gland were as follows: By light microscopic examination, the dilation of small vessels were observed until the 7th day after irradiation. After then, the vascular constriction and decrease in number of small vessels were noticed. Changes were greater on 20Gy irradiated group than on lOGy irradiated group. The reaction to PAS staining at acinar cells was decreased just after irradiation, but gradually recovered with days. There was no specific difference between two irradiated groups. By scanning electron microscopic examination, general findings on the two irradiated groups were similar. The dilation of conduits and meandering were observed on the 3rd day after irradiation. Decrease of capillary density and blunt ended small vessels were appeared on the 7th day after irradiation. After that, findings of the tortuous and twisted vascular running and coarseness of capillary lumen were increased. Changes were greater on 20Gy irradiated group than on l0Gy irradiated group. By transmission electron microscopic examination, increase of the formation of cytoplasmic process was observed on the 3rd day after irradiation. After that, swelling of endothelial cell and bridge formation of cytoplasmic processes were also observed, but destruction of endothelial cell and loss of basement membrane were observed only on 20Gy irradiated group on the 28th day after irradiation.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• v.24 no.1
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• pp.67-77
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• 1994

The purpose of this study was to investigate the irradiatiion effects on the capillary and endothelial cell in the submandibular gland. Sprague-Dawley strain male rats were singly irradiated to their neck region with the dose of 5Gy by 6MV X-irradiation and sacrificed on the 6 hours, 12 hours, 1, 3, 7, and 14days after irradiation. The authors observed the histological changes of the capillary at H & E and PAS staining under a light microscope, and also observed the ultrastructural changes of the endothelial cell using a transmission electron microscope. The obtaining results were as follows: 1. In the light microscopic examination, the capillary density was slightly increased on the 1day after irradiation, and increased until the 7 days after irradiatiion. After then, capillary density was apparently decreased. 2. The reaction to PAS staining at acinar cells was decreased on the 6 hours after irradiation, and recovered on the 7days after irradiation. But reaction was decreased on the 14days after irradiation agan, after then, gradually recovered with days. 3. In the transmission electron microscopic examination, mild proliferation of cytoplasmic process of the endothelial cell and reduction in luminal size were observed just after irradiation. After then, nuclear degeneration, marked proliferation of cytoplasmic process, thickened basal lamina, and numerous cytoplasmic vesicles were observed on the 1day after irradiation. These changes were recovered to normal on the 14days after 5Gy group, but not with 10Gy irradiation group. And destruction of endothelial cell and loss of basal lamina were not observed in both groups. 4. From the above results, reduction in luminal size, proliferation of cytoplasmic process and thickening of basal lamina were observed as the irradiation effects on the capillary and endothelial cell of the submandibular gland. And also, these changes may induce increase in capillary number and endothelial permeability by means of increase of cytoplasmic vesicle formation. The changes appeared earlier and more prominent in 10Gy irradiated group than in 5Gy irradiated group.

• The korean journal of orthodontics
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• v.24 no.2
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• pp.331-348
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• 1994

The purpose of this study was to investigate changes in the shape and ultrastructure of the articular disc of the rat mandibular joint with aging. Mechanical stress applied to the articular disc changes during neonatal, suckling, juvenile, adult and senile stages. Mandibular joints of 6 groups of rats(1-, 7-, 17-, 27-, 55-day and over-1-year groups) were removed en bloc and processed for light and electro microscopic study. The changes in the shape of articular disc were examined by light microscope in each group. Structural and ultrastructural changes in the articular disc were examined by light and electron microscope in each group. The results were as follows : In the 1-day and 7-day groups, the articular disc was long and slender in shape and the articular disc was not fitted with the shape of the mandibular fossa and condyle. However' after that time, the anterior and posterior portions of the articular disc were more bulged and the middle portion was shorter and biconcave. Thus the articular disc was well fitted with the shape of the mandibular fossa and condyle. The cell density decreased with aging. In the l -day and 7-day groups, the Golgi apparatus, rough endoplasmic reticulum and free ribosome, which are involved in the synthesis of intracellular and extracellular matrix, were developed. In the 17-day, 27-day and 55-day groups, not only the cell organelles involved in the synthesis of the intracellular and extracellular matrix but also the cell organelles involved in the remodeling of the extracellular matrix(i.e., finger-like cell process, lysosome and mitochondria)were well developed. With advancing age, intracytoplasmic microfilaments were more accumulated and condroid cells increased. In the over-1-year group, the majority of cells of the articular disc were chondroid cells. The majority of cytoplasmic compartment were filled with intracytoplasmic microfilaments and cell organelles were not developed. Therefore, metabolic activities of the cell was markedly reduced and cells contained structures enduring mechanical stress, and cells which were in the process of degeneration were observed occasionally.

• Clinical and Experimental Pediatrics
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• v.51 no.1
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• pp.54-61
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• 2008

Purpose : This study was designed to clarify the mechanism of proteinuria in nephrotic syndrome patients by using puromycin aminonucleoside (PAN) nephrosis model. Methods : Following administration of various concentrations of PAN and antioxidants we observed the changes of podocyte cytoskeletons in cultured rat glomerular epithelial cells (GEpC) by method of scanning electron microscope, reactive oxyten species (ROS) analysis, permeability assay, confocal microscope, and Western blot assay. Results : PAN not only induced the ultrastructural changes of GEpC, such as shortening and fusion of microvilli, but also separated the intercellular gaps and linear ZO-1. PAN induced oxidative stresses in time and dose dependent manners and increases of intercellular permeability in anti-oxidants inhibitable manners. High concentration of PAN induced not only actin polymerization and disorganization, but also the conglomerulation and internal dislocation of ${\alpha}-actinin$ protein. The intensities of fluorescences of ZO-1 protein were diminished and internalized by PAN in a dose-dependent manner, which were inhibited by anti anti-oxidants. Conclusion : PAN induced the changes of podocytes cytoskeleton and junctional barriers by way of increasing ROS in GEpC that resulted in increasing their permeability in a antioxidatn-inhibitable manner. Glomerular hyperpermeability induced by PAN mediateing through oxidative stresses is thought to take part in the mechanism of proteinuria in nephrotic syndrome.