Showing papers on "Ultrastructure published in 2003"

TEM evidence of ultrastructural alteration on Pseudomonas aeruginosa by photocatalytic TiO2 thin films

Abstract: The antibacterial efficiency of longwave UV-irradiated TiO2 thin films as well as the ultrastructural damage on bacterial cells was evaluated using Pseudomonas aeruginosa as a model. The quantitative antibacterial efficiency assays showed a bacterial inhibition in the range of 32–72% at different times of irradiation. Transmission electron microscopy (TEM) was used to detect the effect of irradiation of TiO2 thin films on the ultrastructure of the bacterial cell in order to reveal possible cellular damage. After 40 min irradiation, an abnormal cellular division was observed: instead of a normal septum, an ‘elongated bridge’ was formed. At a longer irradiation time, wavy structures all around the outer cell membrane were observed, and also some bubble-like protuberances, which expelled inner material. The mechanism of irreversible bacterial cell damage caused by the photocatalytic effect of TiO2 could be related to abnormal cell division, aside from the reported physicochemical alteration of the cell membrane.

Anatomy of Sorgoleone‐Secreting Root Hairs of Sorghum Species

Abstract: Johnsongrass (Sorghum halepense [L.] Pers.) and SX‐17 (Sorghum bicolor × Sorghum sudanese) were investigated microscopically to identify specifically the location of root exudate production. Light, cryoscanning electron, and transmission electron microscopy were used to determine the area of exudate secretion. Light micrographs indicated that the exudate is solely produced by the root hairs. Scanning electron microscopy supported this conclusion. Transmission electron microscopy studies of root hairs support the hypothesis that root exudates are manufactured in the cytoplasmically dense root hair cell in association with smooth endoplasmic reticulum and possibly Golgi bodies. Ultrastructure studies indicated that small globules of cytoplasmic exudate are deposited between the cell wall and the plasma membrane, where they coalesce into larger globules that pass through the cell wall to form droplets near the tip of root hairs.

The ultrastructure of mouse embryonic stem cells.

Abstract: The fine structure of mouse embryonic stem (ES) cell colonies was analysed by scanning and transmission electron microscopy. Most of the ES cells had numerous microvilli of different lengths. Coated pits and vesicles were also seen along areas of the peripheral cytoplasm and plasma membranes. Junctional complexes including gap junctions were observed between adjacent ES cells. These cells had Golgi complexes, spherical to oval mitochondria, lysosomes and typical centrioles, microfilaments and microtubules and large nuclei containing reticulated nucleoli. These results are consistent with the ultrastructural features of undifferentiated cells.

Spermiogenesis and sperm ultrastructure of the liver fluke Fasciola hepatica L., 1758 (Digenea, Fasciolidae): transmission and scanning electron microscopy, and tubulin immunocytochemistry

Abstract: We describe here the ultrastructure of spermiogenesis and the mature spermatozoon of Fasciola hepatica (Trematoda, Digenea, Fasciolidae) by means of scanning (SEM) and transmission electron microscopy (TEM). Spermiogenesis in F. hepatica follows the general pattern of digeneans. However, this is the first report of a flagellar rotation of about 120° in Digenea. The mature spermatozoon of F. hepatica is a filiform cell of about 275 µm in length. SEM and whole mount TEM revealed a helical pattern of axonemes around the sperm body. The spermatozoon presented the same ultrastructural organization as the congeneric F. gigantica. The most interesting features of the male gamete of F. hepatica therefore are the dorsolateral cytoplasmic expansion, the external ornamentation of the cell membrane and the spine-like bodies. We also analysed the distribution pattern of tubulin in the microtubular cytoskeleton of F. hepatica by means of monoclonal anti-tubulins (anti-α -tubulin, anti-β-tubulin, anti-α -acetylated tubulin and anti-α -tyrosinated tubulin). These anti-tubulins labelled axonemal and cortical microtubules but not the central core of the 9+‘1’ trepaxonematan axoneme.

Characterization and behaviour of α‐glucan synthase in Schizosaccharomyces pombe as revealed by electron microscopy

Abstract: α-1,3-Glucan is a cell wall component in Schizosaccharomyces pombe and is exclusive to budding yeast. We analysed the ultrastructure of the cell wall in the α-glucan synthase mutant mok1 and determined the role of α-1,3-glucan in cell wall formation of Sz. pombe. The mok1 mutant cell has an abnormal shape, with swelling at the tip or at the site of the septum. The cell wall is thicker and looser than that of wild-type cells, and the layered structure of the cell wall is broken. The glucan fibrils forming the protoplast retain a fine fibril structure, although their development into bundles is abnormal. We also report the localization of Mok1p by immunoelectron microscopy using high-pressure freeze substitution and SDS-digested freeze-fracture replica labelling methods. The Mok1p is localized on the cell membrane and moves from the cell tip to the medial region during the cell cycle. These results confirm that Mok1p plays an important role in the normal construction of the cell wall and in the primary step of glucan bundle formation, and that it is required for new cell wall synthesis during vegetative growth. These findings suggest that α-1,3-glucan is an essential component for cell wall formation in fission yeast. Copyright © 2003 John Wiley & Sons, Ltd.

Changes in Tomato Leaves Induced by NaCl Stress: Leaf Organization and Cell Ultrastructure

Abstract: The alterations of organization of leaf tissues and cell ultrastructure as a consequence of salt stress (75 and 150 mM NaCl) were studied in two tomato (Lycopersicum esculentum Mill.) cultivars showing different salinity tolerance. The salinity brought changes in cell shape, volume of intercellular spaces and chloroplast number, shape and size. These characteristics were specific in each cultivar. The ultrastructural changes were also different in the two tomato cultivars studied and the most important ones were in the number and size of starch granules in chloroplasts, the number of electron-dense corpuscules in the cytoplasm, the structure of mitochondria, and number of plastoglobuli.

Ultrastructure and behavior of actin cytoskeleton during cell wall formation in the fission yeast Schizosaccharomyces pombe.

Abstract: Fluorescence microscopy has shown that F-actin of the fission yeast Schizosaccharomyces pombe forms patch, cable and ring structures. To study the relationship between cell wall formation and the actin cytoskeleton, the process of cell wall regeneration from the protoplast was investigated by transmission electron microscopy (TEM), immunoelectron microscopy (IEM) and three-dimensional reconstruction analysis. During cell wall regeneration from the protoplast, localization of F-actin patches was similar to that of the newly synthesized cell wall materials, as shown by confocal laser scanning microscopy (CLSM). In serial sectioned TEM images, filasomes were spherical, 100-300 nm in diameter and consisted of a single microvesicle (35-70 nm diameter) surrounded by fine filaments. Filasomes were adjacent to the newly formed glucan fibrils in single, cluster or rosary forms. By IEM analysis, we found that colloidal gold particles indicating actin molecules were present in the filamentous area of filasomes. Three-dimensional reconstruction images of serial sections clarified that the distribution of filasomes corresponded to the distribution of F-actin patches revealed by CLSM. Thus, a filasome is one of the F-actin patch structures appearing in the cytoplasm at the site of the initial formation of the cell wall and it may play an important role in this action.

Infection and propagation of lymphocystis virus isolated from the cultured flounder Paralichthys olivaceus in grass carp cell lines.

Abstract: The causative agent of lymphocystis disease that frequently occurs in cultured flounder Paralichthys olivaceus in China is lymphocystis virus (LV). In this study, 13 fish cell lines were tested for their susceptibility to LV. Of these, 2 cell lines derived from the freshwater grass carp Ctenopharyngodon idellus proved susceptible to the LV, and 1 cell line, GCO (grass carp ovary), was therefore used to replicate and propagate the virus. An obvious cytopathic effect (CPE) was first observed in cell monolayers at 1 d post-inoculation, and at 3 d this had extended to about 75% of the cell monolayer. However, no further CPE extension was observed after 4 d. Cytopathic characteristics induced by the LV were detected by Giemsa staining and fluorescence microscopic observation with Hoechst 33258 staining. The propagated virus particles were also observed by electron microscopy. Ultrastructure analysis revealed several distinct cellular changes, such as chromatin compaction and margination, vesicle formation, cell-surface convolution, nuclear fragmentation and the occurrence of characteristic 'blebs' and cell fusion. This study provides a detailed report of LV infection and propagation in a freshwater fish cell line, and presents direct electron microscopy evidence for propagation of the virus in infected cells. A possible process by which the CPEs are controlled is suggested.

Floral nectary ultrastructure of Prunus persica (L.) Batch cv. Forastero (Newcomer), an Argentine peach

Abstract: Flowers of Prunus persica (L.) Batch. cv. Forastero have an orange toral nectary. The nectariferous tissue was formed by densely packed parenchyma cells (secretory cells) and an epidermis with hairs and modified stomata. The epidermal cells were highly vacuolated with a striated cuticle. The ultrastructure of these cells contained a cytoplasm with endoplasmic reticulum, plastids, mitochondria and dictyosomes. Sub-epidermal cells were barely vacuolated and their ultrastructure was similar to that of the epidermal cells. Differences were observed only in the endoplasmic reticulum, which is organized in a parallel configuration. Plasmodesmata were found between adjacent secretory cells and between secretory and epidermal cells. An electron dense secretion occurred in the intercellular spaces and between the external tangential wall and the cuticle of the epidermal cells. According to the ultrastructural observations, the sugar solution could be passed through the symplast or the apoplast. The nectar could be exuded from the stomata and the micro-channels of the cuticle covering the epidermal cells.

Combined use of confocal laser scanning microscopy and transmission electron microscopy for visualisation of identical cells processed by cryotechniques.

Abstract: Successive visualisation of identical plant cells by light and electron microscopy is reported. For this purpose segments of pea and barley leaves were prepared by high-pressure freezing, freeze-substitution, and low-temperature embedding. The use of Safranin O during low-temperature dehydration allowed, on one hand, staining of all cellular components as investigated by confocal laser scanning microscopy and, on the other hand, excellent ultrastructural and antigenic preservation. A newly constructed specimen holder enabled precise relocation of the target cells for electron microscopic investigations. Transmission electron microscopy and immunohistochemistry revealed that during the whole procedure the ultrastructure of the cells as well as the antigenicity of cell constituents were preserved.

Ultrastructure of spermatogonia and spermatocyte lobules in Taenia solium strobilae (Cestoda, Cyclophyllidea, Taeniidae) from golden hamsters.

Abstract: Golden hamsters (Mesocricetus auratus) were infected with Taenia solium metacestodes dissected from infected pig meat. Adult worms were collected from hamster intestines of animals killed 5-60 days post-infection (dpi), incubated in RPMI 1640 medium with or without colchicine, fixed and processed for transmission electron microscopy (TEM). Sections for light microscopy from 40 different blocks with scolex, immature and mature proglottids were photographed. Thin sections were cut from 25 selected blocks, examined and photographed with TEM. Metaphase mitosis figures were observed in the subtegument of the germinative tissue and interpreted as germ cell precursors. In immature proglottids (20 dpi), discrete cell clusters of three to four cells surrounded by a thin cytoplasmic envelope were identified along the inner border of the lateral excretory ducts. These were also observed in more mature proglottids (40-60 dpi) as clusters of eight cells enclosed in a cytoplasmic enve- lope, with nuclei of spermatogonia exhibiting the synaptolems of primary meiotic cells. In mature pro- glottids from 45 dpi, a large number of spermatocyte lobules were found, exhibiting different stages of spermatogenesis from primary spermatocytes to ma- ture filiform spermatids with a single axoneme, annu- lar nucleus and spiral cortical microtubules, similar to spermatozoa described for type III spermiogenesis of species of the family Taeniidae. All mature spermatocyte lobules were enclosed in a highly orga- nized cellular envelope and surrounded by a basal lamina. The envelopes contained a number of distinct organelles, seen in cross-section as discrete lattices of microtubules located between two layers of plasma membrane, as well as thickened furled cytoplasm with numerous strands of rough endoplasmic reticulum and pockets of microtubules.

Sperm ultrastructure and spermatogenesis in the lizard, Tropidurus itambere.

Abstract: Spermatogenesis, with emphasis on spermiogenesis, is described for the lizard, Tropidurus itambere, using light microscopy, phase contrast and epifluorescence, as well as scanning and transmission electron microscopy. Cellular differentiation involves events of chromatin condensation, nuclear elongation and the formation of structural complexes, such as the acrosomal and axonemal ones. Other new characteris- tics, exclusive for this species, include various aspects of the subacrosomal granule, the insertion of the pro- acrosomal vesicle and the development of these structures to participate in the acrosomal complex. Radial projections occur just above the nuclear shoulders, which have been recognized already from the beginning of cellular elongation. The development of the midpiece, the dense bodies, formation of the flagellum and elimi- nation of residual cytoplasm result in the final characterization of the mature spermatozoon. Comparisons between Tropiduridae and other lizard families are made.

Ultrastructure of the midgut endocrine cells in Melipona quadrifasciata anthidioides (Hymenoptera, Apidae)

Abstract: In this study we describe the ultrastructure of the endocrine cells observed in the midgut of M. quadrifasciata anthidioides. This bee has two types of endocrine cells, which are numerous on the posterior midgut region. Cells of the closed type are smaller and have irregular secretory granules with lower electrondensity than those of the open cell type. The open cell type has elongated mitochondria mainly on the basal area, where most of the secretory granules are also found. Besides the secretion granules and mitochondria, endocrine cells in this species have well-developed autophagic vacuoles and Golgi complex elements.

Light and Electron Microscopy of the Compatible Interaction Between Arabidopsis and the Downy Mildew Pathogen Peronospora parasitica

Abstract: In this study, we focused on compatible interactions between Peronospora parasitica isolate Emoy-2 and wild-type (Oy-0) and mutant (Ws-eds1) Arabidopsis thaliana accessions by using light and transmission electron microscopy (TEM). Light microscopy of compatible interactions revealed that conidia germinated and penetrated through the anticlinal cell walls of two epidermal cells. Rapid spreading of the hyphal growth with formation of numerous haustoria within the mesophyll cells was subsequently followed by profuse sporulation in the absence of host cell necrosis on both wild-type and mutant accessions. TEM observations revealed that coenocytic intercellular hyphae ramified and spread intercellularly throughout the host tissue forming several haustoria in host mesophyll cells. Intracellular haustoria were lobed with the diameter of 6–7 μm. Each haustorium was connected to intercellular hyphae in the absence of apparent haustorial neck. The cytoplasm of the haustorium included the organelles characteristic of the pathogen. Callose-like deposits were frequently observed at sites of penetration around the proximal region of the haustorial neck. Apart from a few callose ensheatments, no obvious response was observed in host cells following formation of haustoria. Most of mesophyll cells contained normal haustoria and the host cytoplasm displayed a high degree of structural integrity. Absence of host cell wall alteration and cell death in penetrated host cell of both accessions suggest that the pathogen exerts considerable control over basic cellular processes and in this respect, response to this biotroph oomycete differs considerably from responses to other pathogens such as necrotrophs.

Renal corpuscle of the sturgeon kidney: an ultrastructural, chemical dissection, and lectin-binding study.

Abstract: The sturgeon is an ancient species of fish that thrives in a wide range of ecological environments, from freshwater to seawater. Basic in this process of adaptation is the ability of the kidney to control fluid filtration and urine formation. However, the morphological basis of this process is mostly unknown. The aim of the present study was to use microdissection techniques (scanning electron microscopy (SEM), transmission electron microscopy (TEM), and lectin-binding histochemistry) to examine the structure of the renal corpuscle of the sturgeon Acipenser nacarii in order to reveal morphologic features that could be related to function, phylogeny, and habitat. The renal corpuscles are aligned along the intrarrenal arteries. The urinary pole shows a siphon-like neck segment (NS) in 92% of the nephrons, whose structural characteristics are different from those of other fish. The podocytes have cuboidal cellular bodies, intercellular contacts, and poorly developed cell processes. The podocyte glycocalyx contains N-acetylglucosamine and lacks sialic acid. The structural and lectin-binding patterns are similar to those found in the immature mammalian kidney. The glomerular basement membrane (GBM) is very thick and consists of three layers: a lamina rara externa, a lamina densa, and a thick subendothelial lamina. The latter contains tubular microfibrils, collagen fibers, and long mesangial cell processes. Frequently, the podocyte bodies attach directly to the GBM, and the area occupied by the filtration slits is very small. Furthermore, the GBM shows a glycosylation pattern different from that observed in most vertebrates. Contrary to what would be expected in sturgeons living in freshwater, the A. nacarii renal corpuscle morphology suggests a low glomerular filtration rate.

Morphology and ultrastructure of Mammillaria gracillis (Cactaceae) in in vitro culture

Abstract: Morphogenetic status of cactus Mammillaria gracillis Pfeiff. tissue culture was studied by light and electron microscopy. In vitro propagated shoots spontaneously developed callus. This callus regenerated normal and hyperhydric shoots without exogenous hormones. Tumour tissue induced by wild or rooty strains of Agrobacterium tumefaciens never expressed any morphogenetic potential. Light microscopy showed cellular characteristics of morphologically different tissues. Ultrastructural studies revealed changes in plastids: secondary dedifferentiation of mature chloroplasts, thylakoid swelling and disruption, phytoferritin accumulation, plastid elongation and increase in size. Changes in chlorophyll and carotenoid content were in accordance with degradation of the thylakoid system. Plastids were confirmed as very sensitive organelles to an artificial hyperhydric environment as well as to Agrobacteria-mediated cell transformation.

Viviparous lizard, Eulamprus tympanum, shows changes in the uterine surface epithelium during early pregnancy that are similar to the plasma membrane transformation of mammals.

Abstract: The “plasma membrane transformation” describes a series of ultrastructural, biochemical, and morphological changes that occur in the uterus of many mammals at the time of blastocyst attachment. These changes, regardless of placental type or length of gestation, include alterations to microvillar length and density and the presence or absence of pinopods or uterodomes. Scanning electron microscopy (SEM) was used to 1) document the topographical ultrastructure of the uterus of Eulamprus tympanum, an eastern Australian viviparous skink with a simple chorioallantoic placenta, for the first time; and 2) determine whether changes identified as “plasma membrane transformation” in mammals occur in E. tympanum. Tissues collected over three seasons from nonreproductive subadult females, preovulatory, postovulatory, and early to mid-gestational females were examined. At low magnification the uterine epithelium of subadults displays a distinctive pattern of tissue folding that includes rectangular areas of tissue delineated by deep lateral and transverse folds. At higher magnification, the uterine epithelium surface is composed of two dominant cell types, i.e., those covered by microvilli and ciliated cells. The folding pattern observed in subadults is less evident in vitellogenic females and the cell surfaces appear highly secretory, with bulging cell apices. Tissue from postovulatory lizards has no distinctive folding pattern and cell surfaces are frequently smooth and lack microvilli. Uterine egg chambers lack ciliated cells at the embryonic pole, but display abundant secretory droplets. Thus, the uterus of E. tympanum undergoes a plasma membrane transformation. The scope of this transformation is not fully understood but may be related to the complexity of placental structure and the development of the embryo/fetus at parturition. J. Morphol. 258:346–357, 2003. © 2003 Wiley-Liss, Inc.

Cytochemical and ultrastructural studies on protoplast formation from disintegrated cells of the marine alga Chaetomorpha aerea (Chlorophyta)

Abstract: Regeneration of protoplasts from extruded cytoplasm and successive development of aplanospores within regenerated cells are described in the marine green alga Chaetomorpha aerea (Dillwyn) Kutzing (Cladophorales, Cladophoraceae). Agglutination of cell organelles in seawater seemed to be mediated by a complementary lectin-carbohydrate system. Three carbohydrates – D-galactosamine, D-glucosamine and α-D-mannose – inhibited agglutination of cell organelles. The presence of these sugar moieties on the surface of cell organelles was verified with their complementary fluorescein isothiocyanate lectins. Agglutination assays using human erythrocytes showed the presence of lectins specific for the above sugars in the protoplasm. The fluorescent probe l-(4-trimethylammoniumphenyl)-6-phenyl-a,3,5-hexatriene revealed that the envelope initially surrounding protoplasts was not a lipid-based cell membrane. Fluorescein diacetate staining showed esterase activity in the protoplasts from the beginning of the regeneration p...

Ultrastructure of smooth muscle, gap junctions and glycogen distribution in Taenia solium tapeworms from experimentally infected hamsters.

Abstract: Taenia solium adults were grown in hamsters infected by feeding them with cysticerci from pig carcasses. Viable strobilae were collected from the hamster duodenum 20-60 days post-infection, fixed and processed for transmission electron microscopy (TEM). Fourteen strobilae were cut into pieces and embedded in individual blocks. Sections, stained with toluidine blue, were then photographed by light microscopy. Over 1,200 TEM images were obtained from selected blocks. Maturing proglottids exhibited a dense myofilament lattice of connecting fibers, each contained in sarcoplamsic extensions of myocytons and emitting cytoplasmic processes loosely attached to other cells, structures characterized as myocyton-myofilament-pseudopod units, which are interpreted as structures involved in the transport of cells and membrane-bound-glycogen from the germinative tissues to mature proglottids. Densely packed membrane-bound glycogen particles were found between the tegumentary cytons of the neck tissue, and as single-stranded particles between the tegumentary cytons of mature proglottids. These were wrapped around cell bodies in the parenchyma of maturing proglottids and as thin cytoplasmic strands between the testicular lobules of mature proglottids. A large number of cell-to-cell adhesions were identified as gap junctions connected to glycogen strands. We suggest that these are involved in the transport of glucose to differentiating tissues.

New findings on sperm ultrastructure of Xenos vesparum (Rossi) (Strepsiptera, Insecta)

Abstract: The systematic position of insect order Strepsiptera is still under debate. It was, therefore, thought of interest to examine the ultrastructure of a strepsipteran in a search for synapomorphies shared with Coleoptera, Diptera, or any other insect order. The fine structure of spermatozoa and the spermatid from Xenos vesparum (Rossi) was re-examined using scanning and transmission electron microscopy and a fixation technique that permits the visualization of the macromolecular organization of the organelles. The spermatozoon was shown to possess several traits that are characteristics of insects in general, such as a 9 + 9 + 2 axoneme, two mitochondrial derivatives containing a crystalline material and two 'zipper lines' present along the sperm tail. Seventeen protofilaments occurred along most of the accessory tubules, which reduced to 16 posteriorly. An acrosome is absent. The neck region contains a prominent centriolar adjunct, which gives rise to two accessory bodies which adhere to the mitochondrial derivatives, and to slender strands of the so-called intertubular material found between the accessory tubules. Of interest is the finding that the glycocalyx consists of prominent filamentous strands, similar to those found in siphonapterans, mecopterans and basal dipterans.

pH-dependent influence of a quaternary ammonium salt and an aminoester on the yeast Saccharomyces cerevisiae ultrastructure.

Abstract: Quaternary ammonium salts inhibited the growth of yeast especially at pH higher (pH 8) than optimal. It was postulated that compounds integrate with the cell membrane and interfere with its functions. The yeast cell ultrastructure investigated under an electron microscope confirms this hypothesis. A relatively high percentage of cells treated at pH 6 with the quaternary ammonium salt of alanine derivative (DMALM-12) at the minimal inhibitory concentration showed an irregularity in the cell shape. No such irregularity was observed in the control. Besides, in the cells treated with the drug, practically no lipid droplets were seen at all. Inside the control cells, electron-dense round bodies were clearly seen and interpreted as vacuoles. These bodies were absent in the cells treated with DMALM-12. Although the yeast cells growing at pH 8 showed a more or less normal shape, they seemed to have difficulty in budding - no fully developed buds were found in the preparations. Only some convexities of the cell wall were seen that could be the beginning of budding which stopped early after the start. Some changes in the round bodies interpreted as vacuoles were visible: they were less dense and full of granules.

Stomach of Aplysia depilans (Mollusca, Opisthobranchia): a histochemical, ultrastructural, and cytochemical study.

Abstract: We report the results of a morphological, histochemical, and cytochemical characterization of the Aplysia depilans stomach, an organ little studied in opisthobranchs. Very thin ciliated cells with microvilli on their apical surfaces are predominant in the epithelium lining the lumen of the stomach. Many lysosomes with a strong arylsulphatase activity were present in the apical regions of these cells that could also contain some lipid droplets and glycogen. Small peroxisomes were observed, usually around lipid droplets or mitochondria. Bottle-shaped secretory cells are very common in this epithelium and produce a secretion rich in proteins and acidic mucopolysaccharides. Most of the cytoplasm of these mucus-producing cells was filled with a very high number of granules and the nucleus is dislocated to the basal region. Cisternae of rough endoplasmic reticulum were abundant around the nucleus and several Golgi stacks were also present in this area. In spite of the variation in the electron density of the granules, only one type of secretory cell seems to be present in the stomach epithelium, since granules with very different electron densities were frequently found in the same cell. A few neurons were also found in the stomach epithelium of this species. Fibrocytes, muscle cells, nerves, and amebocytes were observed in the connective tissue of the stomach wall. J. Morphol. 256:360–370, 2003. © 2003 Wiley-Liss, Inc.

Midgut Ultrastructure of the Third Instar of Dermatobia hominis (Diptera: Cuterebridae) Based on Transmission Electron Microscopy

Abstract: The midgut ultrastucture of the third-instar of Dermatobia hominis (L., Jr.) was investigated using transmission electron microscopy (TEM). The tubular midgut bears a monolayer of epithelial cells with the plasma membrane showing multiple folding where it adjoins the basement membrane. Septate junctions bound the epithelial cells on each side. These cells have electrolucent cytoplasm containing mitochondria, vacuoles, rough and smooth endoplasmic reticula, lamellar bodies, and a prominent nucleus with dispersed chromatin. The peritrophic matrix is close to elongate microvilli, which are sometimes forked. Regenerative cells, in an undifferentiated state when closest to the basement membrane, are scattered throughout the epithelial cells. A thick basement membrane, surrounded by thick connective tissue including muscle, tracheal tubes, and extracellular matrix is linked to epithelial cells by hemidesmosome-like structures. Entero-endocrine, goblet or cuprophilic cells were not observed.

Structural aspects of bulge formation during root hair initiation

Abstract: Using light and electron microscopy, the early stages of root hair initiation were investigated under control conditions and in a situation where F-actin polymerization was effectively inhibited by latrunculin B. Trichoblasts in their early stage of bulge formation possessed large vacuole traversed by cytoplasmic strands and enclosed within a narrow peripheral layer of cytoplasm. The nucleus was settled at the inner periclinal cell wall, typically opposite the site of bulge formation. Within the bulging area, dense cytoplasm and numerous ER elements, and other organelles were accumulated, together with pleiomorphic membrane-bound structures, the identity and nature of which will require further studies. These unusual structures, which were associated with the outer cell wall, contained material similar to that of the cell wall. Similar cell wall-like bodies were observed also in the cytoplasm and sometimes within vacuoles. The possible role of these novel organelles of plant cells in cell wall thinning/degradation or in the turgor pressure maintenance are discussed. Latrunculin B treatment allowed bulge formation but prevented the switch from the slow and diffuse expansion of bulge into the rapid tip-growth characteristic of the emerging root hair. Moreover, the cytoplasm of the bulging domain became extensively vacuolated and lacked abundant ER elements and other organelles including the membrane-bound structures. These results indicate important roles of F-actin in the switch from diffuse to highly polarized tip growth.

Ultrastructure of potato tubers formed in microgravity under controlled environmental conditions

Abstract: Previous spaceflight reports attribute changes in plant ultrastructure to microgravity, but it was thought that the changes might result from growth in uncontrolled environments during spaceflight. To test this possibility, potato explants were examined (a leaf, axillary bud, and small stem segment) grown in the ASTROCULTURETM plant growth unit, which provided a controlled environment. During the 16 d flight of space shuttle Columbia (STS-73), the axillary bud of each explant developed into a mature tuber. Upon return to Earth, tuber slices were examined by transmission electron microscopy. Results showed that the cell ultrastructure of flight-grown tubers could not be distinguished from that of tuber cells grown in the same growth unit on the ground. No differences were observed in cellular features such as protein crystals, plastids with starch grains, mitochondria, rough ER, or plasmodesmata. Cell wall structure, including underlying microtubules, was typical of ground-grown plants. Because cell walls of tubers formed in space were not required to provide support against the force due to gravity, it was hypothesized that these walls might exhibit differences in wall components as compared with walls formed in Earth-grown tubers. Wall components were immunolocalized at the TEM level using monoclonal antibodies JIM 5 and JIM 7, which recognize epitopes of pectins, molecules thought to contribute to wall rigidity and cell adhesion. No difference in presence, abundance or distribution of these pectin epitopes was seen between space- and Earth-grown tubers. This evidence indicates that for the parameters studied, microgravity does not affect the cellular structure of plants grown under controlled environmental conditions.