Kim, Sung-Joo;Kim, Yong-Kuk;Sun, Yuan-Jie;Kim, Soo-Jin;Jeong, Young-Gil;Yu, Yun-Cho;Jo, Seung-Mook
Applied Microscopy
/
v.32
no.4
/
pp.377-383
/
2002
We have detected the murine zinc transporter, ZnT3, and zinc ions in the mouse choroid plexus by immunocytochemistry (ICC) and zinc selenium autometallography ($ZnSe^{AMG}$), respectively. BALB/c mice served as experimental animals. Routine floating ABC immunocytochemical procedures were used for the ZnT3 immunocytochemistry, and the mice were injected intraperitoneally (i.p.) with sodium selenide (10 mg/kg) for the zinc selenium autometallography. The choroid plexus showed weak immunoreactivity (Ir) for ZnT3. At high magnification, ZnT3-Ir was seen to be located in the choroid epithelium and the connective tissue of the capillaries. At the EM level, a high electron density of ZnT3-immunoreactivity was restricted to vesicle membranes as well as microvilli in the apical membrane. In contrast, immunostaining of ZnT3 was completely absent in the basolateral plasma membrane and other cell organelles. After silver enhancement, fine $ZnSe^{AMG}$ grains were observed in both the epithelial and endothelial cells of the choroid plexus. Few $ZnSe^{AMG}$ grains present in the cell bodies of the choroid epithelial cells were located in multivesicular bodies. It is striking that very many $ZnSe^{AMG}$ grains were observed in the endothelial cells of the capillaries. These findings establish the choroid plexus as a non-neuronal pool of zinc ions in the brain, although the functional significance of this pool is not clear. The choroid epithelium, however, may play an important role in the transportation of zinc between the CSF and brain tissue.
Actin and tropomyosin of Cryptosporidium muris were localized by immunogold labeling. Two kinds of antibodies for actin labeling were used. The polyclonal antibody to skeletal muscle (chicken back muscle) actin was labeled on the pellicle and cytoplasmic vacuoles of parasites. The feeder organelle has showed a small amount of polyclonal actin antibody labeling as well. Whereas the monoclonal antibody to smooth muscle (chicken gizzard muscle) actin was chiefly labeled on the filamentous cytoplasm of parasites. The apical portion of host gastric epithelial cell cytoplasm was also labeled by smooth muscle actin together. The polyclonal antibody to tropomyosin was much more labeled at C. muris than host cells, so it could be easily identified even with low magnification (${\times}2,000$). The tropomyosin was observed along the pellicle, cytoplasmic vacuoles, and around the nucleus also. The skeletal muscle type actin seems to play a role in various celluar functions with tropomyosin in C. muris; on the other hand, the smooth muscle type actin was located mainly on the filamentous cytoplasm and supported the parasites firm attachment to host cells. Tropomyosin on the pellicle was thought to be able to stimulate the host as a major antigen through continuous shedding out by the escape of sporozoites or merozoites from their mother cells.
Morphological changes in the uterine and vaginal epithelial cells of the Korean native goats were studied in fifteen primiparous goats slaughtered on the day of parturition and on days 1, 3, 10 and 21 postpartum. 15 uterus and vagina from goats were examined by scanning and transmission electron microscopy. The results obtained in this study were summarized as follows : 1. Transmission electron microscopically, long microvilli which sometimes ramified were found until 10 days postpartum, while short microvilli were found at 21 days. The high electron dense irregular-shaped mitochondria were found in the cytoplasm and the crystalline structure of the mitochondrial matrix was also found from 1 day to 10 days postpartum. Well-developed rough-endoplasmic reticulum (rER) with dilated cisternae which contained the proteins materials was observed at 21 days postpartum. These materials were fused each other and then large granules were found in the free surface of the cytoplasm. A few lipid droplets were generally appeared in the cytoplasm, while numerous droplets were found at 21 days postpartum. A moderate number of ribosomes, a few multivesicular bodies, vesicles, lysosomes and macrophages were found. The globule leucocytes were observed from 0 to 3 days postpartum by transmission electron microscopy. The short microvilli, high electron dense cytoplasm and severe indentation of the nuclear enbelope were found in the vaginal epithelium. Numerouos small vesicles and a few vacuoles were observed in the apical cytoplasmic portion of the epithelium. A few mitochondria were high electron dense and irregular in shape. A moderate amounts of microfilaments, loose intercellular space and dilated rER were also found at 21 days postpartum. 2. Scanning electron microscopically, the folds of the uterine mucosa were generally deep. The long microvilli of the epithelium were found until 3 days postpartum, while short microvili were found at 10 and 21 days postpartum. The distinct intercellular boundary was seen. The apporcine secretory profile of the epithelium observed at between 3 and 10 days postpartum and the cells were somewhat protruded into the lumen. The short microvilli were found on the surface of the protruded cells, while polygonal microridge profile of the epithelium and some dome-shaped epithelium were also observed at 21 days postpartum. The folds of the vaginal mucosa were deep and epithelium was polygonal in shape. The microvilli of the epithelium were long until 3 days postpartum, while they were short at 10 and 21 days. The polygonal epithelium was invaginated into the center of the cell surface until 10 days postpartum. The microridge and dome in shape of the epithelium were found at 10 days postpartum, while the polygonal and exfoliating epithelium were observed at 21 days.
The intestinal tight junction (TJ) plays an important role as a paracellular barrier. Impaired TJ permeability and enhanced proinflammatory cytokine production are crucial pathophysiological mechanisms in inflammatory bowel diseases (IBDs). Although proinflammatory cytokines, tumor necrosis factor-alpha and interluekin-1 beta, which are markedly increased in IBD patients, have been reported to increase intestinal TJ permeability, the role of interleukin-1 alpha (IL-1α) in the TJ has not been studied. Phytochemicals could prevent proinflammatory cytokine-caused TJ alteration. Curcumin (CCM), a biologically active component of turmeric, has a strong anti-inflammatory activity. The purpose of this study was to elucidate the effect of IL-1α on intestinal epithelial TJ permeability and the role of CCM in IL-1α′s action on TJ in an in vitro intestinal epithelial system, Caco-2 monolayers. The TJ integrity of Caco-2 monolayers was estimated by measuring the flux of FITC-labeled dextran and transepithelial electrical resistance (TEER). Apical IL-1α (100 ng/ml) treatment elevated TJ permeability and suppressed TEER of Caco-2 monolayers. Pretreatment with CCM (20 μM) for 30 min significantly inhibited IL-1α-induced TJ alterations, such as increased TJ permeability and decreased in TEER values. These results demonstrated that IL-1α-induced increases in Caco-2 TJ permeability and CCM blocked the action of IL-1α in the TJ.
Background and Objectives : The concentration of sulfur dioxide($SO_2$) gas in the ambient air appears increasing in the industry and urban area day by day. It was known that $SO_2$ is noxious gas. $SO_2$ can be irritating to the eyes, nose, throat, upper respiratory tract and skin. It produces sulfurous acid on contact with water and is extremely irritating to the nasopharynx and respiratory tract. Laminin is a family of extracellular matrix glycoproteins localized in the basement membrane that separates epithelial cells from the underlying stroma. The biological activities of laminin are to promote cell migration, wound healing, growth and differentiation. Meterials and Methods : The histologic changes and the expression of laminin in tracheal mucosa sacrificed at every weeks (to 7 weeks) after continued $SO_2$ exposure of 250ppm for 30 minutes a day were studied in rats. Results : Pathologic tissue was formed at the tracheal mucosa and the underlying tissue by the infiltration of monocytes and epithelium was transformed to the single cell layered epithelium above 5 weeks after exposure. At the 6 weeks after exposure, epithelial cells were partially lost and epithelial cell layer was transformed to be leaf-shaped. Submucosal tissue was transformed to be lymphatic tissue. An intense positive staining for laminin was found in apical cytoplasm and lateral surface of the normal epithelial cells and basement membrane but at the 5 and 6 weeks after exposure, laminin activity was decreased to the moderate activity. At the 7 weeks after exposure, laminin activity was decreased to the weak activity. Conclusion : Our finding suggests that $SO_2$ makes histologic damage on the tracheal mucosa and decreases immunoreactivity for laminin. Longer duration of the exposure of $SO_2$ makes more histologic damage on the tracheal mucosa and decreases immunoreactivity for laminin.
Han, Kyong Ha;Li, Zhun;Kang, Byeong Jun;Youn, Joo Yeon;Shin, Hyeon Ho
Korean Journal of Environmental Biology
/
v.37
no.1
/
pp.31-41
/
2019
The toxic dinoflagellate Gymnodinium catenatum isolated from the southern coast of Korea was described under light and scanning electron microscopy, and its large subunit (LSU) rDNA was sequenced. In addition, the effects of temperature and salinity on its growth were investigated. The cells of G. catenatum, as viewed under the electronic microscope, were green-brown color, $38.1-77.4{\mu}m$ in length and $26.1-40.8{\mu}m$ in width. The epicone was conical, while the hypocone was trapezoidal. The nucleus was located at the central part of the cell. The apical groove was horseshoe-shaped and small pores were irregularly distributed on the cell surface. Molecular phylogeny based on LSU rDNA gene sequences showed that the Korean G. catenatum and previously reported species formed a monophyletic clade within Gymnodinium sensu stricto clade. The maximum growth rate of $0.37day^{-1}$, was obtained at $25^{\circ}C$ and 35 psu, and the maximum cell density of $1,073cells\;mL^{-1}$, was observed at $20^{\circ}C$ and 25 psu. However, G. catenatum did not grow at temperature < $15^{\circ}C$ and < $30^{\circ}C$. These results suggest that environmental conditions of summer and autumn in the southern coast of Korea may be favorable for the growth of G. catenatum.
Flow cytometry (FCM) was used to measure the ploidy level of three different sports from 'Campbell Early' ($Vitis$$labruscana$) grape. Results of the study showed different ploidy levels. FCM analysis for 'Campbell Early' grape which contains 2C DNA diploid cells showed single peak around 35-40 while 'Kyoho' grape with 4C DNA tetraploid cells had a different level of 70-80. However, analysis of the sports displayed a histogram with 2 peaks containing both 2C and 4C nuclei. There was no difference in histograms of 2C DNA flesh and pericarp; on the other hand, 4C DNA flesh type of sports had a different histogram from that of the 2C DNA pericarp. Chromosome numbers of diploid ('Campbell Early'), tetraploid ('Kyoho'), and three sports were counted under the microscope. 'Campbell Early' and 'Kyoho' have 38 and 76 chromosomes, respectively. Three different sports are mixoploids with mixtures of diploid and tetraploid cells. Microscopic observations of shoot apical meristems in sports from 'Campbell Early' grape were carried out to determine the type of plant chimera. 'Campbell Early' grape (diploid) and 'Kyoho' grape (tetraploid) showed that both had 2 tunica layers covering corpus cells, while the three different sports had tunica layers showing mostly oblique division. Most cells from 'Kyoho' grape were larger than 'Campbell Early' grape. Cells from L-2 and L-3 layers of the three sports were similar to 'Kyoho' grape in size, although all cells in L-1 surface layer were uniform in size like 'Campbell Early' grape. Results of FCM analysis indicated that both normal and polyploid cells could be intermixed in sports and could become mixoploidy consisting of diploid and tetraploid. All sports used in the tests were periclinal chimera plants with two distinct L-1 and L-2 cell layers. The result of this study suggests that all three sports which originated from 'Campbell Early' grape might be 2-4-4 type chimera formation.
Jo, Seung-Mook;Kim, Sung-Jun;Park, Seung-Kook;Kang, Tae-Cheon;Won, Moo-Ho
Applied Microscopy
/
v.30
no.4
/
pp.357-365
/
2000
Paneth cells have been suggested to contribute to the elimination of excess metals into the intestinal lumen. The purpose of this study wat to investigate the changes of the zinc pools in rats subjected to functional loading with zinc salt by mean of both light and electron microscopical autometallography (AMG). Wistar rats 4 were administrated with zinc chloride (20 mg/kg body weight) intraperitoneally dissolved in 1 ml distilled water. The control group received 1 ml saline IP. After further one hour the animals were transcardially perfused with 0.4% sodium sulphide dissolved in 0.1 M PB fellowed by 3% glutaraldehyde solution for 10 minutes. Pieces of ileum were frozen with solid $CO_2$ and sectioned on a cryostat. The sections $(20{\mu}m)$ were autometallographically developed. Sections selected for EM were reembedded on top of a blank Epon block, from which ultrathin sections (100 nm) were cut. The ultrathin sections were double stained with uranyl acetate (30 min) and lead citrate (5 min), then examined under electron microscope. Studies of comparable sections from control and zinc loaded animals with the AMG selenium method gave quite different results. The control animals demonstrated a weakly positive staining in the cytoplasm of the Paneth cells. In the electron microscope the AMG silver grains were found to be located in the cytoplasm, while the electron dense secretary granules and other cell organelles were void of staining. Few AMG grains were located at the apical surface of the Paneth cells. In sections from zinc loaded rats, the AMG grains were seen in abundance in the lumen of the Lieberkuhn crypts at light microscopic levels. At EM levels the zinc revealing silver grains were located in the cytoplasm as in the controls, but much more AMG grains were shifted into the secretary granules. Furthermore, profound AMG grains were found in the lumen of the crypts and surrounding vessels. And a few grains were seen in the endothelium. The AMG technique demonstrated a pattern of AMG grains in the Paneth cells that strongly suggests a transport of zinc ions through these cells.
Hur, Jingyung;Choi, Eunseok;Buckley, Kenneth J.;Lee, Sukchan;Davis, Keith R.
Molecules and Cells
/
v.26
no.2
/
pp.131-139
/
2008
Expression of the seven open reading frames (ORFs) of single-stranded DNA Curtoviruses such as Beet curly top virus (BCTV) and Beet severe curly top virus (BSCTV) is driven by a bi-directional promoter. To investigate this bidirectional promoter activity with respect to viral late gene expression, transgenic Arabidopsis plants expressing a GUS reporter gene under the control of either the BCTV or BSCTV bi-directional promoter were constructed. Transgenic plants harboring constructs showed higher expression levels when the promoter of the less virulent BCTV was used than when the promoter of the more virulent BSCTV was used. In transgenic seedlings, the reporter gene constructs were expressed primarily in actively dividing tissues such as root tips and apical meristems. As the transgenic plants matured, reporter gene expression diminished but viral infection of mature transgenic plants restored reporter gene expression, particularly in transgenic plants containing BCTV virion-sense gene promoter constructs. A 30 base pair conserved late element (CLE) motif was identified that was present three times in tandem in the BCTV promoter and once in that of BSCTV. Progressive deletion of these repeats from the BCTV promoter resulted in decreased reporter gene expression, but BSCTV promoters in which one or two extra copies of this motif were inserted did not exhibit increased late gene promoter activity. These results demonstrate that Curtovirus late gene expression by virion-sense promoters depends on the developmental stage of the host plant as well as on the number of CLE motifs present in the promoter.
Morphological changes of the epithelium of the endometrium by prolonged treatment of $17{\beta}$-estradiol or progesterone in ovariectomized rats was studied at the ultrastructural level. The epithelium of the endometrium in ovariectomized rats was characterized by the appearance of a number of vacuoles which was contained with the membraneous structures, lipid droplets and the others. The epithelium was low cuboidal, and a few short microvilli were present at the cell surface. Secretory granules are rarely found. After estradiol treatment, the epithelium was high columnar in shape. The mitochondria was appeared throughout the cytoplasm, however, long or swelling mitochondria was often found. Golgi apparatus and rER were relatively well-developed. Relatively long and sparse microvilli were present at the cell surface. After progesterone treatment, the epithelium was characterized by the appearance of numerous vesicles at the apical region and numerous lipid droplets at the subnuclear region. At the cell surface a number of short and blunt microvilli were found. These data indicated that the endometrium was dependent on estrogen and progesterone for changes in both its morphological and functional state and suggested that each hormone exerted a unique effect on the epithelial cells.
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