• Applied Microscopy
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• v.26 no.1
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• pp.47-57
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• 1996

This study analysed the transport properties of bladder mucosa known as the typical system of 'tight epithelia' by using TEM observation with both rapid freeze-fracture electron microscopy and thin-section method and mainly analysed the cellular characteristics of turtle bladder epithelial cells. The bladder epithelium, like other tight epithelia, consists of a heterogenous population of cells. The majority of the mucosal cells are the granular cells and may function primarily in the process of active $Na^+$ reabsorption in turtle bladder. The remaining two types of cells are rich in mitochondria and is believed to be res-ponsible for a single major transport system, namely, $H^+$ transport by A-type of cell and urinary $HCO_{3}^-$ secretion by B-type of cell. As viewed in freeze-fracture electron micrograph, the tight junctions form a continuous tight seal around the epithelial cells, thus restricting diffusion in tight epithelia. In addition, the apical surface membranes have a population of rod-shaped intramembranous particles (IMPs). It is believed that these IMPs probably represent the components of the proton pump. However, it is likely that these characteristics of the apical transporter remain to be clarified in tight epithelial cells.

• 한국전자현미경학회:학술대회논문집
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• 2004.05a
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• pp.55-56
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• 2004

• Applied Microscopy
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• v.28 no.1
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• pp.83-90
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• 1998

The pathway of intracellular transport of dehydrocholic acid was investigated in the hepatocytes of rats by transmission electron microscopy with conventional and freeze fracture methods. Both in normal and experimental groups, the cis Golgi cisterns were sacculated and faces toward the bile canaliculus. In the experimental group, however, the cis Golgi cisterns showed buds, which were probably separated to be vesicles. Some of the buds were connected to the cisterns with the narrow neck. The vesicles were increased in the vicinity of bile canaliculi. The fusion between vesicles and bile canaliculus were frequently observed in the experimental group. This was particularly well shown in the freeze fracture replica. In the thin section, the vesicles were devoid of visible contents as seen in the bile canaliculli. The evidence suggests that the vesicles are derived from the cis Gogi cistern in the way that buds pinch off, serve as vehicles to transport dehydrocholic acids and fuse to bile canaliculi for exocytosis.

• Applied Microscopy
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• v.16 no.2
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• pp.1-13
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• 1986

The purpose of this study was to investigate the morphology and intercellular junctions of the odontoblast of dental pulp in the rat incisor by means of the freeze fracture electron microscopy. Twenty male Sprague-Dawley rats weighing $150{\sim}200g$ were used. After being anesthetized by an intraperitoneal injection of 0.5 ml sodium pentobarbital per kg in body weight(60 mg/ml) the animals were perfused with 2.5% glutaraldehyde-2% paraformaldehyde fixative in 0.1 M cacodylate buffer, pH 7.2 through the ascending aorta for one hour. The incisors were carefully extracted from the jaws and demineralized by suspending them in 0.1 M EDTA in 3% glutaraldehyde (pH 7.2) for two weeks. After demineralization, the specimens were obtained from the portion divided into five equal parts. For freeze-fracture replication, demineralized tissues were infiltrated for several hours with 10%, 25% glycerol in 0.1M cacodylate buffer as a cryoprotectant and then frozen in liquid Freon 22 and stored in liquid nitrogen. Fracturing and replication were done in Balzers BAF 400D high-vacuum freeze-fracture apparatus at $-120^{\circ}C$ under routine $5X10^{-7}$ Torr vacuum. The tissue was immediately replicated with platinum unidirectionally at $45^{\circ}$ angle and reinforced with carbon at $90^{\circ}$ angle unidirectionally or by using a rotary stage. The replication process was monitored by a quartz-crystal device. The replicas were immersed in 100% methanol overnight. The tissue was then digested from the replica by clorox (laundry bleach), placed into 5% EDTA, and washed repeatedly with distilled water. The replicas were picked up on 0.3% formvar-coated 75 mesh grids and examined in the JEOL 100B electron microscope. The results were as follows; 1. Both in thin sections and freeze-fracture replicas, three types of intercellular junctions were recognizable in the plasma membrane of odontoblast: gap junction, tight junction and desmosome-like junction. 2. The nuclear pores were evenly distributed over the nuclear envelope. The pore complex formed a ring about 70 nm in diameter. 3. Gap junctions were found between odontoblasts as well as odontoblasts and neighbouring pulp cells (fibroblast, subodontoblastic cell process, nerve-like fibre). Gap junctions, which were round, ellipsoid and pear-shaped and 600 nm in diameter, were observed in the odontoblast. 4. Numerous round and ellipsoid gap junctions could be frequently seen on the plasma membranes in cell body and apical part of the odontoblasts. On the P face, the junctions were recognized as a cluster of closely packed particles, measuring about 9 nm in diameter, and on the E face, the junctions were recognized as a shallow grooves.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.21 no.1
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• pp.38-52
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• 1995

The liposomes have been developed in many drugs and cosmetics fields. In this context, it should be mentioned that MLV liposomes can be prepared standing with the main compounds of the intercellular lipids, cholesterol, palmitic acid, cholesteryl ester and lecithin, by swelling reaction. We report properties of the formation of MLV liposomes with use of the lipid base and Microfluidizer. MLV liposomes formed as multilamellar vesicles(MLV). The effect of the gelation of MLV liposomes have been on swelling reaction which have been mixed lipid with polyol and water phase at high temperature(90$\pm$5$^{\circ}C$). MLV liposomes have been prepared in incorporating alpha hydroxy acid ligrediens. Optimum condition of MLV liposomes were passed three times in the microfluidizer, particle size of the vesicles should be within 150-350nm and those confirmed by freeze-fracture electron microscopy.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.1
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• pp.123-128
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• 2004

Numerous novel ingredients have been introduced for the higher functionality of whitening cosmetics. Through the preliminary research, we have found Trichosanthes kirilowii root, Prunella vulgaris leaf and Clematis chinensis root have high whitening efficacy. But they are insoluble. Moreover the discoloration of and decrease in content take place when they are exposed to light, heat or oxygen. From Trichosanthes kirilowii root, Prunella vulgaris leaf and Clematis chinensis root, efficacious ingredients were ethanol-extracted by heating to 75∼85$^{\circ}C$ for 6∼8 h. These extracts have the inhibitory activity of tyrosinase and B16 melanin formation, thus enhancing whitening effect. We made liposomes using propylene glycol (PG)/hydrogenated lecithin/middle chain triglycerides (MCT)/glycerin/water and microfuidizer to stabilize extracts. The stability against heat and light was enhanced by 3∼5 times compared with untreated extracts. Particle size analyzer, freeze fracture transmission electron microscopy (FF-TEM), chromameter and HPLC are used for the analysis.

• Applied Microscopy
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• v.25 no.2
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• pp.53-64
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• 1995

This study was designed to investigate the morphological alterations of zonula occludens, macula adherences and gap junctions between the hepatocytes in the fasting conditions. Animals (Sprague Dawley, $250{\sim}280g$) were divided into two groups: normal and fasting. The latter were fasted for eight days prior to sampling. Liver tissues were sectioned and replicated after freeze fracturing for the transmission electron microscopy. In the normal rat liver, the interhepatocellular space at the area of some zonula occludens appeared to be widened in thin sections. On the freeze fracture replicas., the zonula occludens appeared as an anastomosing network of $2{\sim}4$ strands or grooves on P or E faces. Free ends and fragments of the strands were observed. In the rat fasted for eight days, the hepatocytes were diminished in size and the organelles were decreased in number and size. The intercellular space was wide at many areas of zonula occludens in thin section. On the freeze fracture replicas, the zonula occludens showed diminution or disappearence of anastomosing network of strands or grooves. Free ends and small fragments of the strands or grooves were frequently encountered. The macula adherens was markedly increased in number in thin sections, although they could not be found on the freeze fracture replicas. The gap junctions were increased in number in thin sections. Small aggregations of the intramembranous particles appeared with larger ones on the freeze fracture replica. The evidences may suggest the followings: (1) The disassembly of zonula occludens in the fasting states is led from the diminished mechanical stress on the luminal surface of bile canaliculus with the impaired secretion of bile components from the hepatocytes. (2) The increase of macula adherens is necessary to maintain the liver parenchyma integrity in the fasting state which leads the hepatocyte to be diminished and finally the intercellular space to be separated. (3) The rise in both number and size of gap junctions is owing to the need of increasing intercellular communication between the hepatocytes during the fasting. (4) The alteration of zonula occludens is easily led by the physiological condition of hepatocytes even in the normal ones.

• Applied Microscopy
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• v.22 no.2
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• pp.84-96
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• 1992

This study was carried out to examine the $H^+$ transport mechanism by observing the properties of cellular membrane having an ${\alpha}$ type of carbonic anhydrase (CA)-containing cells in turtle urinary bladder. The urinary bladder consists of a heterogenous population of cells. As a result of fine observation with traditional thin-section electron microscopy. the bladder epithelium has three different cell types on mucosal surface. They are a basal cell, a granular cell and a third type of CA-rich cell. The CA-rich cells are divided into two distinct smaller groups within them and called them ${\alpha}$ type and ${\beta}$ type of CA cells. The ${\alpha}$ type of CA cells are responsible for the proton secretion using the proton pumps on the apical plasma membrane, while the ${\beta}$ type of CA cells secrete bicarbonate via an oppositely-directed proton pumps in their basolateral plasma membrane. After performing the freeze-fracture technique, it was shown that there were distributed a large number of intramembranous particles having a special structure on the apical membrane of ${\alpha}$ type of CA-rich cells in the process of their $H^+$ secretion. In turtle bladder ${\alpha}$ type of CA-rich cells, this particle was the only prominent structure in the apical membrane. These intramembrane rod-shaped particles probably represent the integral membrane components of the proton pump. This result may explain that carbonic anhydrase within epithelial cell of urinary bladder takes part in formation of $H^+$ and bicarbonate, that active transport of $H^+$ is done, and that the reabsorption of bicarbonate suggests transport mechanism containing $H^+$ secretion. However, it seems that more studies are required for considering their regular transport pathway.

• Applied Microscopy
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• v.19 no.2
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• pp.119-137
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• 1989

It has been shown in this and earlier investigation that the turtle bladder mucosa has three main cell types on their mucosal surface. They are the granular cells, ${\alpha}$ CA cells, and ${\beta}$ CA cells. The three major transport mechanisms that occurs in the turtle bladder are sodium reabsorption, proton secretion, and bicarbonate secretion. In the present work the trans-port mechanisms by bladder epithelial cells of freshwater turtle, Pseudemys scripta, are summarized as follows. 1. The granular cells play an important role in sodium transport, while the ${\alpha}$ and ${\beta}$ CA cells do not appear to play a determining role in sodium transport. 2. It appears that the active sodium transport in the granular cells occurs in two-step process, implying that first, sodium diffuses into the cells, followed by an energy-dependent efflux step, which is catalyzed by the ouabain-sensitive Na-K ATPase. 3. The ${\alpha}$ type of CA cells are responsible for the proton secretion using the proton pump on the apical plasma membrane, while the ${\beta}$ type of CA cells are believed to be responsible for bicarbonate secretion. 4. When looked at under freeze-fracture electron microscopy, the apical plasma membrane of ${\alpha}$ cells have a characteristic population of rod-shaped intramembranous particles which are believed to be components of the proton pumps. Conversely, ${\beta}$ type of CA cells show rod-shaped particles in their basolateral plasma membranes, which is consistent with the proton absorptive, bicarbonate secretory mechanism. 5. In the turtle bladder, the ${\alpha}$ and ${\beta}$ type of cells are believed to be both responsible for proton transport, but in opposite directions.