The morphological changes of the midgut epithelium during the metamorphosis of pine caterpillar are observed with light and electron microscope, being divided into 5 stages from the 8th instar larva to just after pupation. The midgut epithelium of 8th instar larva is composed of columnar cell, goblet cell, regenerative cell, and endocrine cell. The secretorials are arranged on the nuclear membrane in the columnar cell of the midgut epithelium in the 8th instar larva, and lysosomes are augmented in the apical portion. Cytoplasmic extrusions are observed in the apical surface of columnar cell but they have no cell organells. Nucleus, mitochondria, rER, Golgi complex, and free ribsomes are observed in the regenerative cell. Regenerative cells are differentiated into the form of goblet cell, and vacuoles are gradually increased in the cytoplasm. Just pupa stage, the materials, which appears to be mainly composed of Ca, are observed in the circular form and goblet cavity of regenerative cell are detached to lumen. As a result, it reflects the process of the degeneration of the midgut epithelium that lysosomes are gradually augmented in the columnar cell, that nuclear materials are removed to cytoplasm, and that cytoplasmic extrusions are observed in the apical surface. And though regenerative cells are differentiated into the form of the goblet cell, it is believed that goblet cavity is detached from regenerative cell to the lumen and midgut epithelium of pupa stage is formed.
This investigation was undertaken to establish the ultrastructural organization of the retina in domestic fowl (Gallus domesticus B.) comparing with the ultrastructure that has been indicated in other Aves by several workers. The electron microscope observations were made on selected segments of retinal tissue prefixed for 2 hrs in 1.25% glutaraldehyde buffered with 0.2 M cacodylate at pH 7.2 and then postfixed in cold 1% osmium tetroxide in 0.4 M cacodylate buffer for 2 hrs. After postfixation, tissues were dehydrated in alcohol series, embedded in Epon 812 mixture from propylene oxide and stained with saturated uranyl acetate and $Pb(NO_3)_2$ solution. Specimens were examined with a Hitachi HS-7S electron microscope. The pigment epithelia cells contain numerous mitochondria with prominent dense granules and several changeful spaped Golgi bodies. The internal fine structure of the receptor outer segments revealed the characteristic stacks or arrays of bimembranous disks. The ellipsoid outer portion of the cone inner segments is composed of a tightly packed mass of extraordinarily large mitochondria. The outer limiting membrane is seen to contain many junctional complexes, the fibrillar material of which is electron-dense.
The morphogenesis of neuroblasts and plexiform layers, and establishment of its synapses were studied by electron microscopy in human embryos and fetuses ranging from 10 mm to 260 mm crown-rump length ($5\sim30$ weeks of gestational age). At 30 mm fetus the developing retina was composed of outer and inner neuroblastic layers . Cell division of outer neuroblast was occurred until 90 mm fetus. The transient layer of Chievitz was formed by 30 mm fetus, inner plexiform layer by 50 mm fetus, and outer plexiform layer by 150 mm fetus. The cytoplasm of differentiating ganglion cells contained ribosomes, rough endoplasmic reticula, Golgi complexes, microtubules and dense bodies. The processes of $M\ddot{u}ller$ cell penetrated between groups of ganglion cell axons, and formed the cellular component of the inner limiting membrane at 30 mm fetus. At 90 mm fetus radial fibers of M ller cells contained extensive smooth endoplasmic reticula and microtubules. In each specimen , apposing paired membrane specializations were classified as junctions without synaptic vesicles, conventional synapses and ribbon synapses. At 50 mm fetus the processes of neuroblasts in inner plexiform layer were interconnected by junctions without synaptic vesicles. Conventional synapses developed by addition of synaptic vesicles to initially vesicle-free junctions at 90 mm fetus. At 150 mm fetus ribbon synapses were first recognized by the inclusion of a prominent electron-dense material associated with synaptic vesicles. By 260 mm fetus conventional and ribbon synapses and junctions without synaptic vesicles formed similar to those found in the adult.
Park, Choon K.;Kwan H. You;Young K. Deung;Lee, Choon K.;Ho S. Chung
The Korean Journal of Zoology
/
v.25
no.1
/
pp.9-28
/
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.
The spermiogenesis of a Korean octopus, Octopus minor, inhabiting western of Korea Sea was observed by electron microscopy . The obtained results are as follows: The spermiogenesis of Octopus miner proceeds through four stages; early- , mid- , and late-spermatid, and mature sperm. An early spermatid is a spherical cell looking light due to the low electron density. The acrosome formed from Golgi complex of the upper nucleus looks dark due to the high electron density. The extra-nuclear rod (enr) stemming from proximal centriole is transformed from round shape into oval shape, elongating to the upper nucleus. In our observation, the axoneme was being formed from distal centriole, and the manchette composed of a number of microtubules is also found around nuclear membrane. In a mid-spermatid, chromatins in the nucleus contract shaping fine threads, and the manchette is also observed around nuclear membrane. Especially, the spherical acrosome is transformed into long oval one which is tinged with a number of horizontal stripes and has the middle electron density. In a late-spermatid, chromatins in the nucleus contract thick and short. Furthermore, the mitochondrial sleeve, in which the axoneme is surrounded with mitochondria, is observed at middle piece. The axoneme has a typical structure of 9+2 and around it, 9 coarse fibers are observed. Also in the acrosome cavity of mature sperm, horizontal striation is found. However, regularly spaced processes are peculiarly observed in there. A sperm is about 390 fm long, whose head is bent a little like a banana while the acrosome region is helical. In the middle piece of sperm, $11\sim12$ mitochondria are surrounding coarse fibers that reach the main piece of tail, while nothing but 9+2 structured axoneme is found in the end piece.
The morphological and fine structural changes during the oogensis of Clonorchis sinensis were studied on the developing ovums in the ovary and ootype with electron microscope. Adult worms were removed from the hepar of the which and previously infected with metacercariae of Clonorchis sinensis. The ovary including the Mehlis' glands and an ootype from adult worm was prefixed for 1-2 hours in 1.25% glutaraldehyde buffered with 0.2M cacodylate at PH 7.2, secondarily fixed for 30 minutes in potassium bicromate and postfixed for an hour in 1% osmic acid buffered with 0.4M cacodylate at PH 7.2. After fixation tissues were dehydrated in an alcohol series, embedded in Epon 812 from propylene oxide and stained with saturated uranyl acetate and $Pn(NO_3)_2$ solution. Material was examined with a Hitachi HS-7S electron microscope. The periphery of the ovary, except for the posterior region, is made up of oogonia. As the oogonia divide they proliferate primary oocytes toward the central part of the ovary. After a period of growth the primary oocyte leaves the ovary and is penetrated by a sperm in the ootype. Sperm penetration immediately activates the primary oocyte to resume its meiotic activity. After the oocytes meiotic activity is completed, the pronuclei fuse to form a single cleavage nucleus which possesses two nucleoli. As the oocytes develop their cytoplamic materials are abundant; small mitochondria are abundant and often their profiles are more unmerous in one part of the cytoplasm than elsewhere; the granular endoplasmic reticulum becomes alveolar-sac form after it leaves the ovary it becomes stratified form. The reticulate Golgi apparatus is apparent in the developed oocyte. A little of cortical granules are distributed inside of the plasma membrane I oogonia and large quantity of cortical granules are arranged just inside of the plasma membrane of the primary oocyte and after fertilization they are disappeared with broken out.
This study was performed to investigate the distribution and differentiation on the immunoreacted cells of the ChAT (choline acetyltransferase) at the Meynert basal nucleus of the forebrains in the growth periods of rat, using the immunohistochemistric method. According to the cell shape and the ratio of long axis vs short axis of cell soma, the ChAT antibody reacted nerve cells in the Meynert basal nucleus of the rats were classified into six types. In the adult rats, the FD (frequency distributions) of round, oval and elongated cells were maximum. The FD of these types were shown to be progressively decreased during the postnatal development. In addition, the FD of elongated nerve cells in were observed in the adult rats respectively. This was thought to be the same phenomenon as those in the round and oval cells . The total mean volume of ChAT antibody reacted cell somata was lowest in the PND (postnatal days) 7 rats and was highest in the PND 21 rats. But, those were decreased to the adult. These results suggest that ChAT antibody reacted nerve cells grow up to PND 21 and then, differentiate into the various types by neurites outgrowing. On the electron micrography, the adult rat forebrain cells were obtained to be well developed ribosomes, polysomes , rough endoplasmic reticula and mitochondria. The immunreactivities were observed in ribosomes, polysomes, rough endoplasmic reticula and outer membrane of mitochondria. Golgi complexes were poorly developed and not showed jmmunreactivity. The ribosomes , polysomes and endoplasmic reticula are considered to be closely related to the inter cellular localization and biosynthesis of the ChAT but not Golgi complex. According to the results in the present study, it is considered that the ChAT-immunoreacted nerve cells in the Meynert basal nucleus of the rat forebrains are differentiated throughout the postnatal development with following processes of changes; 1) the cholinergic nerve cells develop postnatally 2) cell soma volumes gradually increase during the early postnatal days 3) and then, cells differentiate into the various types by projecting the neurites to the appropriate area after PND 21.
Kim, Woo-Kap;Park, Hong-Duok;Kim, Eun-Soo;Han, Sung-Sik
Applied Microscopy
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v.9
no.1
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pp.57-69
/
1979
The ultrastructural changes of embryo and endosperm cells were observed during the green fruit with embryo about $250{\mu}$ long to germination. 1. In the embryo cells of green fruit with embryo about $250{\mu}$ long, mitochondrial cristae and plastid are undifferentiated and dictyosome are occasionally observed. There are electron-opaque globoids in the vacuole and a lot of spherosomes in the outer layer of smooth endoplasmic reticulum. Endosperm is filled with spherosomes and electron-opaque protein bodies surrounded by spherosomes, and due to these, other organelle are not observed. 2. In the embryo cells of seeds with red seed coat, mitochondrial cristae are well developed, electron-opaque globoids increased, and vacuoles are enlarged. In the endosperm, however, spherosomes increased, protein bodies are enlarged, and electron-opaque globoidal crystals are dispersed within them. 3. In the procambium and epicotyl cells of dehiscent seed, Golgi vacuoles and vesicles are well developed, and mitochondrial cristae are also well differentiated. Spherosomes are numerously present and radicle cells, peripheral cells of hypocotyl, and vacuoles of cotyledon are well differentiated. Endosperm is filled with spherosomes containing electron-opaque granules and protein bodies are surrounded by a single membrane. There are acid phosphatase around globoids and spherosomes. 4. At the time of seeding, spherosomes markedly increased in the outer layer of cotyledon and protein bodies are also observed. Cell organelles are differentiated and plastids containing starch are also present. 5. In the outer $2{\sim}3$ layers of cotyledons, radicle cells, and peripheral cells of hypocotyl during post-seeding to germination, spherosomes and plastids with starch increased, and mitochondria and microbodies are also found around the nucleus of embryo cells. With approaching, the germination stage, in the endosperm contacting with embryo, vacuoles are well differentiated but spherosomes decreased. There increased electron-opaque materials within vacuoles. In other endosperm, with the decrease of spherosome, mitochondria increased and electro n-opaque globular bodies are formed and gradually increased. The outer layer of protein bodies are reduced while electron-transparent portions are enlarged and fused together to occupy the outer layer where small particles are formed. 6. In the endosperm of germination stage, spherosomes decreased while protein bodies, are fused together to form 2 or 3 within a cell.
This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.
We have investigated the action of sodium nitrite on the growth and morphologic changes of T uaginolis and on the treatment of subcutaneous abscess by trichomonad in mice. Sodium nitrite inhibited the growth of metronidazole-sensitive KT9 isolate and metronidazole-resistant CDC85 strain of T vcsinalis as concentration of 6 mM and 10 mM respectively Intraperitoneal injection of sodium nitrite (70 ㎍, 100 ㎍, 130 ㎍/g body weight) did not reduce the size of abscess produced by subcutaneous inoculation of T uasinnlis in mice. T uosinnlis, treated with sodium nitrite at concentration giving about 50% inhibition of growth, showed fissures, many vacuoles and electron-translucent zone in the cytoplasm by transmission electron microscopy. In the case of CDC85 treated with 9 mM sodium nitrite, hydrogenosomal matrical change , destruction of hydrogenosomal membrane, autophagic vacuoles, disappearance of Golgi complex and polysome were notably observed . With above results, it is assumed that sodium nitrite inhibits the growth of metronidazole-sensitive and - resistant strains of T. ucsinalis and induces the morphological changes of 7 uusinalis although it does not affect in reducing of abscess size by vagiginalis in mice.
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