Functional ultrastructure of the midgut of the fire ant Solenopsis saevissima Forel 1904 (Formicidae: Myrmicinae)

Solenopsis saevissima has a midgut composed of columnar, regenerative, and goblet cells. The midgut epithelium was covered by a basal lamina. Outside the basal lamina, layers of inner oblique, circular, and outer longitudinal muscles were present. Columnar cells showed a basal plasma membrane containing numerous folds, mitochondria, and the nucleus. Rough endoplasmic reticulum, Golgi bodies, membrane bounded vacuoles, and spherocrystals were found in this region. The apical plasma membrane was constituted by microvilli, which were above a region rich in mitochondria. Regenerative cells were found in groups lying by the basal lamina. Goblet cells were associated with an ion-transporting mechanism between the haemolymph and the midgut epithelium. These cells were lying by the midgut lumen and large microvilli were evident, but the cytoplasmic features were similar to the columnar cells.     

取食转Bt基因水稻对稻纵卷叶螟幼虫中肠的组织病理学效应

利用透射电镜观察了稻纵卷叶螟 Cnaphalocrocis medinalis（Guenée）幼虫取食转Bt基因水稻后中肠的组织病理变化。结果表明：稻纵卷叶螟幼虫取食转cry1Ab基因水稻后,中肠上皮细胞的线粒体先发生形态变化,随连续取食时间的延长线粒体出现凝聚、内嵴稀疏、空泡化等,在后期还呈凝聚态随突起脱落或沿杯腔边沿单一排列。内质网的变化也很明显,病变过程中伴随着粗糙内质网的肿胀、核糖体脱落,粗糙内质网增多等现象。细胞核的变化较小,在处理后期出现细胞核拉长、核仁聚集等变化。组织病变程度不一,有的细胞在病变早期就出现了空泡化。     

A new enveloped helical virus from the blue crab,Callinectes sapidus

Quite different ultrastructural changes were observed in the columnar cell and the goblet cell of the silkworm midgut after administration of the crystalline toxin of Bacillus thuringiensis. Shortly after the ingestion of the toxin, the deep infoldings of the basal cell membrane of some columnar cells became very irregular in shape and the mitochondria near the basal region were transformed into a condensed form. A few goblet cells showed relatively high electron density in the cytoplasm. The earliest pathological changes were slight and located in a region lying between the first and second thirds of the midgut. With the passage of time, they spread anteriorly and posteriorly to include the entire anterior two thirds of the midgut and became more profound. The cytoplasm of columnar cells became very electron transparent. Most mitochondria were transformed into a condensed form and the endoplasmic reticulum assumed a vacuole-like configuration. The basal infoldings of the cell membrane almost disappeared. On the other hand, the cytoplasm of the goblet cells became very electron dense and granular. The clear basal infoldings of the cell membrane were enlarged making a striking contrast with the dense cytoplasm. However, the mitochondria and the endoplasmic reticulum did not show any pathological deformation.     

Isolation of separate apical,lateral and basal plasma membrane from cells of an insect epithelium. A procedure based on tissue organization and ultrastructure

The tissue used in this study was the midgut of the tobacco hornworm larva, Manduca sexta. The midgut epithelium is a single layer of cells resting on a thin basal lamina and underlying discontinuous muscle layer. The epithelial cells are of two main types, goblet and columnar cells, joined together by the septate junctions characteristic of insect epithelia. From this tissue we were able to isolate four distinct plasma membrane fractions; the lateral membranes, the columnar cell apical membrane, the goblet cell apical membrane and a preparation of basal membranes from both cell types. The lateral membranes were isolated by density gradient centrifugation following gentle homogenization of the midgut hypotonic medium, which caused the cells to rupture at their apical and basal surfaces, releasing long segments of lateral membranes still joined by their septate junctions. For isolation of apical and basal membranes the tissue was disrupted by ultrasound, based on the light microscopic observation that carefully controlled ultrasound can be used to disrupt each cell in layers starting at the apical surface. The top layer contained the columnar cell apical membrane, which consists of microvilli forming a brush border covering the lumenal surface of the epithelium. The second layer contained the goblet cell apical membrane, which is invaginated to form a cavity occupying the apical half of the cell, and the third layer contained the basal membranes. As each layer was stripped off the epithelium it was collected and the plasma membrane purified by differential or density gradient centrifugation. For all four membrane fractions, the isolation procedure was designed to preserve the original structure of the membrane as far as possible. This allowed electron microscopy to be used to follow each step in the isolation procedure, and to identify the constituents of each subcellular preparation. Although developed specifically for M. sexta midgut, these techniques could readily be modified for use on other epithelia.     

Zur Cytologie der Rectaldrüsen von Knorpelfischen

The stratified epithelium of the central collecting duct of the elasmobranch(Scylliorhinus canicula, Galeorhinus galeus andRaja batis) rectal gland consists of 3 to 6 layers of cells: one superficial, and several basal cell layers. In the superficial layer normally three different types of cells can be distinguished (a) goblet cells, (b) cells with apical secretory granules and (c) flask-shaped cells. The superficial layer ofScylliorhinus canicula reveals a further cell type, so-called mitochondria-rich cells. The epithelial areas built by these cells are always single-layered. The goblet-cells are very similar to goblet cells found in the intestine of vertebrates. Their dominant structures are a well developed ergastoplasm, a large Golgi-apparatus and mucous granules compactly filling the apical cell region. The cells with apical secretory granules are columnar or dumbbell shaped. They contain a rough-surfaced endoplasmic reticulum and a well developed Golgi-apparatus. The secretory granules are loosely distributed within the Golgi-field and are arranged in one or more rows just below the cell apex. The flask shaped cells are characterized by a cytoplasm rich in small vesicles. They posses few dictyosomes and several small mitochondria. There is some evidence for endocytotic activity. The mitochondria-rich cells are characterized by lateral cell interdigitations, by a basal labyrinth and by numerous mitochondria. They are similar to the excretory cells of rectal gland parenchyma. The cells of the basal epithelium layers are differenciated only to a small extent. They are joined in a loose formation with white blood cells often found in the intercellular spaces. The function of the elasmobranch rectal gland is not restricted to the excretion of concentrated salt solutions. There is also a significant secretion of mucous substances. The tubule glands are primarily excretory, the epithelium cells of the central collecting duct mainly secretory in function.     

Cell differentiation in the embryonic midgut of the tobacco hornworm, Manduca sexta

While the larval midgut of Manduca sexta has been intensively studied as a model for ion transport, the developmental origins of this organ are poorly understood. In our study we have used light and electron microscopy to investigate the process of midgut epithelial cell differentiation in the embryo. Our studies were confined to the period between 56 and 95 hr of embryonic development (hatching is at 101 hr at 25 degrees C), since preliminary studies indicated that all morphologically visible differentiation of the midgut epithelium occurs during this time. At 56 hr the midgut epithelium is organized into a ragged pseudostratified epithelium. Over the next 10 hr, the embryo molts and the midgut epithelium takes on a distinctive character in which the future goblet and columnar cells can be identified. With further differentiation, closed vesicles in the goblet cells expand and subsequently communicate to the outside by way of a valve. The columnar cells form numerous microvilli on their apical surfaces that extend over the goblet cells. Both cell types form basal folds from a series of plasmalemmal invaginations. Differentiation occurs concurrent with a six-fold elongation of these cells.     

The ultrastructure of the posterior midgut caecum of Pachygrapsus crassipes (Decapoda, Brachyura) adapted to low salinity

The effects of salinity adaptation and of composition and tonicity of fixatives upon the ultrastructure of the posterior midgut caecum (PMC) of Pachygrapsus crassipes have been studied. The PMC epithelium consists of a single layer of columnar cells with a microvillous border. The apical cytoplasm contains numerous mitochondria, lysosomes, and much smooth endoplasmic reticulum. Rough endoplasmic reticulum and Golgi apparatus are situated in the perinuclear cytoplasm. This epithelium resembles other transporting epithelia in that the basal cytoplasm has an extensive system of branched tubules formed from invaginations of the lateral and basal plasma membrane. Numerous mitochondria are associated with the basal tubular system. To determine the possible contribution of the PMC to the osmoregulatory ability of Pachygrapsus, the ultrastructure of the PMC from animals adapted to 40, 50, 100 and 150% sea water was investigated. Enlargement of basal tubules and intercellular spaces at low salinity, suggestive of fluid-transport activity, was found to be an artifact of fixation. The most consistent response when animals were acclimated to dilute salinities was that some basal mitochondria assume a more complex shape, usually appearing as rings in cross sections of the caecum. A hypothesis concerning the functional significance of these mitochondria is proposed.     

Electron microscopic study of the anterior midgut in Cenocorixa bifida Hung. (Hemiptera: Corixidae) with reference to its secretory function

The epithelium of anterior midgut of adult Cenocorixa bifida was examined with light and electron microscopy. The folded epithelium is composed of tall columnar cells extending to the lumen, differentiating dark and light cells with interdigitating apices and regenerative basal cells in the nidi surrounded by villiform ridges that penetrate deeply into the epithelium. The columnar cells display microvilli at their luminal surface. Microvilli lined intercellular spaces and basal plasma membrane infoldings are associated with mitochondria. These ultrastructural features suggest their role in absorption of electrolytes and nutrients from the midgut lumen. The columnar cells contain large oval nuclei with prominent nucleoli. Their cytoplasm is rich in rough endoplasmic reticulum, Golgi complexes and electron-dense secretory granules indicating that they are also engaged in synthesis of digestive enzymes. The presence of secretory granules in close proximity of the apical plasma membrane suggests the release of secretion is by exocytosis. The presence of degenerating cells containing secretory granules at the luminal surface and the occurance of empty vesicles and cell fragments in the lumen are consistent with the holocrine secretion of digestive enzymes. Apical extrusions of columnar cells filled with fine granular material are most likely formed in response to the lack of food in the midgut. The presence of laminated concretions in the cytoplasm is indicative of storageexcretion of surplus minerals. The peritrophic membrane is absent from the midgut of C. bifida.     

Histopathological effects of Bacillus thuringiensis on the midgut of tobacco hornworm larvae (Manduca sexta): Low doses compared with fasting

The cytology and ultrastructure of the midgut cells of Manduca sexta larvae are described for untreated controls, larvae which fed on a spore preparation of Bacillus thuringiensis, and larvae which were fasted for either 24 or 48 hr. New observations on the ultrastructure of midgut cells in Manduca larvae included the finding of specialized Golgi vesicles in anteriormost columnar cells and of regular arrays of expanded rough endoplasmic reticulum in goblet cells of the posterior midgut region. The present observations reveal that the columnar cells of the midgut responded cytologically in the same way to fasting as they did to exposure to the toxic spores of B. thuringiensis. The goblet cells, however, appeared unaffected by fasting but became swollen in response to feeding of B. thuringiensis spore preparation.     

Cytological and enzyme-histochemical investigations on the digestive organs of Nautilus pompilius (Cephalopoda, Tetrabranchiata)

The foregut, stomach, caecum, midgut, and rectum of the digestive tract of Nautilus pompilius L.were investigated with ultrastructural and enzyme-cytological methods. Three different cell types were identified within the lamina epithelialis mucosae: main cells, goblet cells, and cells with secretory granules. The main cell type is the epithelial cell with microvilli, a basal nucleus surrounded by dictyosomes, rough endoplasmic reticulum, mitochondria, and electron-dense granules identified as lysosomes in the apical part of the cell. In the caecum this cell type contains endosymbiotic bacteria. The presence of endocytotic vesicles and the storage of lipids in the caecum indicate that this organ is involved in the process of absorption. In the caecum and the longitudinal groove of the rectum the main cells are, in addition, ciliated, facilitating the transport of food particles and faeces. Two types of goblet cells are found in all organs except in the stomach, forming a gliding path for food particles and protecting the epithelium. In the foregut and rectum, cells with electron-dense granules were recognized as the third type. The conspicuous secretory cells of the rectum represent a delimited rectal gland; its possible biological function is discussed. The tunica muscularis in all organs of the digestive tract consists of obliquely striated muscle cells innervated by axons containing transparent, osmiophilic and dense-cored vesicles. Positive reactions for acid and alkaline phosphatase, monoamine oxidase, β-glucuronidase, and trypsin- and chymotrypsin-like enzymes are localized in the lamina epithelialis mucosae.     

Digestive cells in the midgut of Triatoma vitticeps (Stal, 1859) in different starvation periods

Triatoma vitticeps (Stal, 1859) is a hematophagous Hemiptera that, although being considered wild, can be found in households, being a potential Chagas’ disease vector. This work describes the histology and ultrastructure of the midgut of T. vitticeps under different starvation periods. Fifteen adults of both sexes starved for 3, 7, 20 and 25 days were studied. In general, digestive cells had apical microvilli, basal plasma membrane infoldings and central nucleus. The perimicrovillar membrane was found in all insects examined. Digestive cells of anterior midgut had lipid droplets, glycogen granules, developed basal labyrinth associated with mitochondria suggesting their role in nutrient storage and in fluid and ion transport. The cells of median and posterior regions of the midgut were rich in rough endoplasmic reticulum, lysosomes, vesicles and granules with different electron-densities. Moreover, cells of the posterior portion of the midgut had hemozoyn granules and mitochondria in the apical cytoplasm close to microvilli, suggesting their role in blood digestion and active nutrient absorption. The midgut of T. vitticeps showed differences in digestive cells associated with the time after feeding, and the increase of vesicles amount in long starvation periods, which suggests enzyme storage, which is readily used after a blood meal.     

The structure of the chambered nautilus siphuncle: The siphuncular epithelium

The siphuncle of the chambered nautilus (Nautilus macromphalus) is composed of a layer of columnar epithelial cells resting on a vascularized connective tissue base. The siphuncular epithelium taken from chambers that have not yet begun to be emptied of cameral liquid has a dense apical brush border. The great number of apical cell junctions (zonula adherens) compared to the number of nuclei suggests extensive interdigitation of these cells. The perinuclear cytoplasm of these preemptying cells is rich in rough endoplasmic reticulum. The siphuncular epithelium of both emptying and “old” siphuncle (which has already completed emptying its chamber) both show little rough endoplasmic reticulum but do contain extensive systems of mitochondria-lined infoldings of the basolateral plasma membranes. Active transport of NaCl into the extracellular space of this tubular system probably entrains the water transport involved in the chamber-emptying process. Both emptying and old siphuncular epithelium also show large basal infoldings (canaliculi) continuous with the hemocoel, which appear to be filled with hemocyanin. The apical cell junctions of emptying and old siphuncular epithelium contain septate desmosomes that may help to prevent back-flow of cameral liquid into the chambers.     

Goblet cell membrane differentiations in the midgut of a lepidopteran larva.

So-called goblet cells are present in the midgut of lepidopteran larvae. They are thought to be involved in the active transport of potassium out of the haemolymph and into the gut lumen. A number of plasma membrane differentiations within the goblet cell cavity has been investigated using conventional staining, lanthanum tracer and freeze-etch techniques. Of particular interest are junction-like inter- and intra-membrane differentiations found on the villus-like cytoplasmic projections present at the apical tip of the goblet cell cavities. These cytoplasmic projections appear to act as a valve; in some cases they seem to close off the top of the goblet cell cavity, so isolating it from the gut lumen, while in other cases they are spread apart leaving a wide channel from the cavity into the lumen. The junction-like structures on these cytoplasmic projections are different in structure from the septate-type junctions which seal the midgut cells together at their apical borders, and the 2 types are present on the same plasma membrane, often within one micron of each other. The need for a different type of junction may possibly be related to the fact that it occurs between 2 areas of the same plasma membrane. The morphology of this unusual junction-like structure is discussed and 2 diagrams are presented to illustrate our interpretation of its structure.     

Bacillus thuringiensis crystal toxin: Ultrastructural studies of its effect on silkworm midgut cells

Bacillus thuringiensis crystal toxin induced a cytoplasmic response in columnar cells within 1 min after ingestion although external symptoms were not exhibited by larvae until 15 min after ingestion. Microvilli became less consistently uniform in diameter; their organized internal microfilaments were disrupted and disappeared. The cisternae of rough endoplasmic reticulum were enlarged and denuded of ribosomes. By 5 min after ingestion, microvilli of some columnar cells disappeared entirely and gross ultrastructural changes were observed in other regions of the cells. Up to 5 min after ingestion there were few, if any, ultrastructural changes observed within goblet cells. Mitochondria in columnar cells were swollen but did not exhibit the condensed configuration reported by other workers. Both the buffer system used in the fixation medium and its osmolarity influenced the changes in the ultrastructure of midgut cells exposed to B. thuringiensis crystal toxin.     

Replacement of midgut epithelium in the greater wax moth,Galleria mellonela,during larval-pupal moult

The epithelium of larval midgut of the greater wax moth, Galleria mellonela, was replaced during the larval-pupal moult. The development of this moth was tentatively divided into 11 stages, from the full-grown larva of last instar to the 4-day-old pupa. The midgut at each stage was observed for (1) overall structure, (2) the position of goblet cells, and (3) the appearance of the yellow body. Light microscopy revealed that cell death in the midgut began in a cocoon-spinning larva (stage II), when pigments in the stemmata started to migrate. Before drastic remodeling started to occur, cytoplasmic projections in the goblet cavities were transformed. The larval midgut changed markedly at stage III, when the pigments left the stemmata. The epithelium of the larval midgut dropped as a whole into the lumen, transforming into the yellow body. Simultaneously, a pupal midgut epithelium developed. Electron microscopy of the columnar cells of a stage III larva showed that microvilli and mitochondria looked normal even though the nucleus with condensed heterochromatin resembled an apoptotic nucleus of vertebrate and higher plant cells. Caspase-3-like protease activity was restricted to the larval midgut and increased in parallel with the formation of the yellow body. The results indicate that the replacement of the larval midgut is facilitated by a typical apoptotic process.     

The ultrastructural investigation of the midgut in the quill mite Syringophilopsis fringilla (Acari,Trombidiformes: Syringophilidae)

The midgut of the females of Syringophilopsis fringilla (Fritsch) composed of anterior midgut and excretory organ (=posterior midgut) was investigated by means of light and transmission electron microscopy. The anterior midgut includes the ventriculus and two pairs of midgut caeca. These organs are lined by a similar epithelium except for the region adjacent to the coxal glands. Four cell subtypes were distinguished in the epithelium of the anterior midgut. All of them evidently represent physiological states of a single cell type. The digestive cells are most abundant. These cells are rich in rough endoplasmic reticulum and participate both in secretion and intracellular digestion. They form macropinocytotic vesicles in the apical region and a lot of secondary lysosomes in the central cytoplasm. After accumulating various residual bodies and spherites, the digestive cells transform into the excretory cells. The latter can be either extruded into the gut lumen or bud off their apical region and enter a new digestive cycle. The secretory cells were not found in all specimens examined. They are characterized by the presence of dense membrane-bounded granules, 2–4 μm in diameter, as well as by an extensive rough endoplasmic reticulum and Golgi bodies. The ventricular wall adjacent to the coxal glands demonstrates features of transporting epithelia. The cells are characterized by irregularly branched apical processes and a high concentration of mitochondria. The main function of the excretory organ (posterior midgut) is the elimination of nitrogenous waste. Formation of guanine-containing granules in the cytoplasm of the epithelial cells was shown to be associated with Golgi activity. The excretory granules are released into the gut lumen by means of eccrine or apocrine secretion. Evacuation of the fecal masses occurs periodically. Mitotic figures have been observed occasionally in the epithelial cells of the anterior midgut.     

Growth and differentiation of the larval midgut epithelium during molting in the moth, Manduca sexta

The number of epithelial cells comprising larval midgut of the tobacco hornworm moth, Manduca sexta increases 200-fold in development from the first to the fifth instar. We have examined larvae periodically before and during molting to follow epithelial cell proliferation and differentiation. The midgut epithelium in Manduca sexta consists predominantly of columnar and goblet cells. These are arranged in a characteristic pattern with each goblet cell surrounded by a single layer of 4-6 columnar cells (Hakim et al., (1988)). While undifferentiated basal stem cells are infrequently seen in intermolt larvae, just prior to the period when external signs of molting are visible, their number increases and mitotic figures become common. Proliferation continues for several hours and then these stem cells differentiate following a pattern similar to that seen during embryogenesis (Hakim et al., (1988)). Here, however, the newly differentiating cells become intercalated among the mature differentiated cells already present in the epithelium. Since the pattern of individual goblet cells surrounded by a reticulum of columnar cells is maintained after the addition of new cells, the midgut epithelium of molting larvae appears to be a useful model for studying pattern formation in development.     

Regeneration of midgut epithelial cell in the silkworm, Bombyx mori, infected with viruses

In the larvae of the silkworm, Bombyx mori, the regeneration of midgut cells infected with a cytoplasmic polyhedrosis virus (CPV), a flacherie virus (FV), and a small DNA virus (SDV) was studied. Large numbers of newly developed cells appeared in the CPV-infected part of the midgut epithelium just before larval molt, and along with their development, the CPV-infected old columnar cells were discharged into the midgut lumen during the molt. On the other hand, in the uninfected portion of the midgut only a few cells developed, and no columnar cells were discharged. Similarly, the marked replacement of midgut epithelial cells during larval molt was also observed in larvae infected with CPV + FV. In the larvae infected with CPV + SDV, the columnar cells lost their regenerative ability, and because of the exfoliation of infected columnar cells, the midgut epithelium consisted mainly of uninfected goblet cells at a late stage of infection. The degree of epithelial regeneration varied with the silkworm strain and the dosage of the virus.     

Structural and ultrastructural features of boar seminal vesicles

The morphological features of boar seminal vesicles were examined by light and transmission microscopy. Boar seminal vesicles consist of glandular tissue arranged in multiple lobules containing a system of ramified secretory tubules. The secretory tubules are composed of a mucosa formed by an epithelium and an underlying lamina propria and, are surrounded by a muscular layer. The epithelium is made up of columnar cells and occasional basal cells. Mast cells are frequently found among epithelial cells. Three types of columnar cells, considered different stages of the secretory cell cycle, are present: principal cells, clear cells and dense cells. Principal cells are functionally differentiated cells characterised by abundant mitochondria, great development of the rough endoplasmic reticulum and presence of secretory granules in their cytoplasm. The apical surface of many principal cells shows apical blebs filled with PAS-positive material. No acid mucosubstances are detected. Microvilli cover the apical surface except in the apical blebs. Dense cells, arranged between principal cells, are also functional differentiated cells but with signs of cellular degeneration. Clear cells are an initial differentiated stage of columnar cells and are characterised by the presence of a poorly developed rough endoplasmic reticulum and by the absence of secretory granules. Proliferating cells are present among columnar cells. Basal cells contain scarce cytoplasm, few organelles and no secretory granules. The lack of mitotic activity in these cells suggests that they do not act as precursors of columnar cells.     

STUDIES ON SMALL INTESTINAL CRYPT EPITHELIUM : I. The Fine Structure of the Crypt Epithelium of the Proximal Small Intestine of Fasting Humans

Small intestinal crypt epithelium obtained from normal fasting humans by peroral biopsy of the mucosa was studied with the electron microscope. Paneth cells were identified at the base of the crypts by their elaborate highly organized endoplasmic reticulum, large secretory granules, and small lysosome-like dense bodies within the cytoplasm. Undifferentiated cells were characterized by smaller cytoplasmic membrane-bounded granules which were presumed to be secretory in nature, a less elaborate endoplasmic reticulum, many unattached ribosomes and, in some cells, the presence of glycogen. Some undifferentiated cells at the base of the crypts contained lobulated nuclei and striking paranuclear accumulations of mitochondria. Membrane-bounded cytoplasmic fragments, probably originating from undifferentiated and Paneth cells, were frequently apparent within crypt lumina. Of the goblet cells, some were seen actively secreting mucus. In these, apical mucus appeared to exude into the crypt lumen between gaps in the microvilli. The membrane formerly surrounding the apical mucus appeared to fuse with and become part of the plasma membrane of the cell, suggesting a merocrine secretory mechanism. Enterochromaffin cells were identified by their location between the basal regions of other crypt cells and by their unique intracytoplasmic granules.