A tubular configuration of the granular endoplasmic reticulum forming a raft-like parallel array in the pinealocytes of two species of Japanese moles (Mogera kobeae and M. wogura)

Summary Ten or more straight tubules, each of which consists of a double unit membrane of granular endoplasmic reticulum with a cylindrical profile, are joined side by side in a raft-like configuration in the cytoplasm of the pinealocytes of Japanese moles. They measure about 60 nm and 100 nm in their inner and outer diameters, respectively, and are often partially connected to unspecialized granular endoplasmic reticulum. Cisterns held between the inner and outer unit membranes with cylindrical profiles vary from 15 nm to 30 nm in width. Ensheathed portions of the cytoplasm are contiguous with cytoplasm outside the tubular units. The inner unit membranes of the tubules bear fewer ribosomal particles than the outer ones.     

Morphometric differences in midgut epithelial cells between strains of female Aedes aegypti (L.) (Insecta,Diptera)

Midgut epithelial cells of female Aedes aegypti deriving from 2 different mosquito strains were compared morphometrically. One of the 2 strains was recently isolated from nature (East Africa). The other strain is an old laboratory strain, reared under laboratory conditions for about 30 years. The quantitative morphological comparison demonstrates, that the ultrastructural composition of the midqut epithelial cells corresponds generally in both strains. It can be shown, however, that in the old strain, compared to the new strain, the organelle amount of midgut epithelial cells is significantly reduced during the first 53 days of the mosquito's life. Parallel to decreased relative volumes of organelles and reduced surface densities of membrane systems an increase in lysosome-like structures is observed. These observations are interpreted as deterioration processes, possibly due to the long rearing in the laboratory. Therefore, care should be taken in the selection of an A.aegypti strain for any quantitative morphological or physiological investigation.     

Morphometric comparison of the midgut epithelial cells in male and female Aedes aegypti L. (Insecta, Diptera).

Midgut epithelial cells of male and female Aedes aegypti, 3 days after emergence, were compared morphometrically. The results of the present investigation concerning the female, are in good agreement with those of a previous study (Hecker et al., 1974), demonstrating that morphometric investigation of midgut epithelia in A. aegypti can successfully be reproduced, and that the mosquito strain used did not show quantitative morphological changes due to laboratory rearing. In males, the cells of the anterior (A) and posterior part (P, 'stomach') of the midgut differ in their quantitative composition. Higher values are found for the microvilli and for the basal labyrinth in the A-part. On the other hand a higher volume density of the mitochondria is present in the P-part. No significant differences are found in the A-part between males and females. Significant differences, however, are present in the P-part. Distinctly more rer in the female stomach can be correlated with the synthesis of enzymes for blood digestion, which are absent in the male. In addition, the more complex functions of the female P-part are also reflected by higher values for other organelles and membrane systems (e.g. mitochondria, basal labyrinth).     

Morphometric parameters of the midgut cells of Aedes aegypti L. (Insecta,Diptera) under various conditions

Summary Previous morphometric or biochemical investigations have yielded different data on the distribution of free and membrane-bound ribosomes in midgut cells of Aedes aegypti. In the present paper ribosomal distribution has been morphometrically analysed to determine whether different mosquito strains, different food and different narcosis used in these previous studies, and/or methodological errors, could account for the different results.Most of the cellular parameters in the stomach epithelium of female A. aegypti, strain Rockefeller, and their changes during blood digestion, are comparable to those measured for another Aedes strain (Segemaganga, Hecker and Rudin 1979), and are generally similar to those of Anopheles stephensi (Hecker 1978). Proteolytic activity against casein is similar for both Aedes strains with a maximum activity being registered around 30 h after a blood meal. During digestion of human serum there is no increase in the ratio of membranebound to free ribosomes, and no significant increase in the surface area of the rough endoplasmic reticulum or of the number of bound or free ribosomes. Proteolytic activity is distinctly lower than during blood digestion. Immobilization of mosquitoes prior to dissection by ether narcosis or by shaking in a test tube has no significant influence on cellular parameters in females fed on sugar solution and investigated 3days after emergence.It is concluded that the differences in ribosomal parameters previously obtained by morphometrical (Hecker and Rudin 1979) and biochemical (Gander et al. 1980) methods, can only partly be explained by the selection of different food for the mosquitoes, and must also have been caused by methodological inadequacies.

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Zusammenfassung Frühere morphometrische und biochemische Untersuchungen erbrachten teilweise unterschiedliche Resultate betreffend Verteilung freier und membrangebundener Ribosomen in Mitteldarmzellen von Aedes aegypti. In der vorliegenden Arbeit wurde morphometrisch untersucht, ob diese Unterschiede bedingt waren durch die Verwendung verschiedener Mückenstämme, unterschiedlichen Futters und verschiedener Narkosemethoden durch die beiden Arbeitsgruppen, oder ob methodische Einflüsse dafür verantwortlich waren.Die meisten Zellparameter im Magenepithel von A. aegypti, Stamm Rockefeller, wie auch ihre Änderungen während der Verdauung eines Blutmahls, entsprachen den für einen andern Aedes-Stamm (Segemaganga, Hecker und Rudin 1979) gemessenen Werten und stimmten im allgemeinen mit denjenigen für Anopheles stephensi (Hecker 1978) überein. Die proteolytische Aktivität gegen Casein war bei beiden Stämmen gleich mit einem Aktivitäts-Maximum um 30h nach Blutmahl. Bei der Verdauung von menschlichem Serum konnte keine Zunahme des Verhältnisses von membrangebundenen zu freien Ribosomen, keine signifikante Oberflächenvergrößerung des rauhen endoplasmatischen Retikulums und keine signifikante Erhöhung der Zahl gebundener und freier Ribosomen gemessen werden. Die Proteaseaktivität war deutlich schwächer als während der Verdauung von Blut. Betäubung der Mücken vor der Sektion mit Aether oder durch Schütteln in Reagenzgläsern ergab im Vergleich keinen signifikanten Einfluß auf die Zellparameter von Zuckerwasser-gefütterten Weibchen, die drei Tage nach dem Schlüpfen untersucht wurden.Unterschiede in den Ribosomenparametern, die mit morphometrischen Methoden (Hecker und Rudin 1979) einerseits und biochemischen (Gander et al. 1980) andererseits untersucht wurden, konnten nur teilweise durch die Wahl unterschiedlichen Futters für die Mücken durch die beiden Arbeitsgruppen erklärt werden. Es müssen zusätzlich methodische Einflüsse für diese Unterschiede verantwortlich sein.

     

Ultrastructural dynamics of the corpus allatum of Choleva angustata Fab. (Coleoptera,Catopidae)

Summary The ultrastructure of the corpora allata (CA) during postembryonic stages of Choleva angustata Fab. shows cyclic changes, in particular regarding the endoplasmic reticulum and the mitochondria. During the last larval instar, at the short transitory period which follows the cessation of alimentation, we observed a highly unusual appearance of the CA, characterized by intensive vacuolization. After this transitory period, the smooth endoplasmic reticulum (ser) disappears until the imago emerges. These observations along with our experimental data permit a physiological interpretation.We thank Departments of Electron Microscopy and Photography of the Institute of Neurophysiology and Psychophysiology, C.N.R.S., Marseille, France     

Functional morphology of the midgut of Aedes aegypti L. (Insecta,Diptera) during blood digestion

Morphometric analysis of the epithelial lining of the stomach of A. aegypti suggests that digestion of the first blood meal in the stomach of this species can be viewed as a series of phases that can be correlated with physiological data from the literature. In phase Ia (0-10 h after blood meal [abm]) the whorls of the rough endoplasmic reticulum unfold, the Golgi zones increase, and the basal labyrinth is enlarged. This coincides with processes of synthesis and secretion (e.g., peritrophic membrane, esterases and lipases) and transport by the stomach epithelium. In phase Ib (10-20 habm) the cellular parameters measured further increase, indicating high synthetic and secretory activities (e.g., digestive enzymes). In phase Ic (20-30 habm) cell structures involved in synthesis and secretion still exhibit high values coinciding with maximal activity of proteases in the gut. Enhanced surface area of microvilli, prominent lipid inclusions, and appearance of glycogen deposits in the gut epithelium suggest increased absorption, storage, and transport functions of the stomach cells. In phase II (30-36 habm) structural alteration points to a gradual shift from synthesis and secretion to absorption, partial storage, and transport of nutrients. In phase III (36-72 habm) the cellular apparatus is reduced concomitant with the ending of the digestive cycle. Lipid inclusions and glycogen deposits disappear from the stomach epithelium.     

Normal versus alpha-amanitin induced cellular dynamics of the midgut epithelium in female Aedes aegypti L. (Insecta, Diptera) in response to blood feeding.

In midgut epithelial cells (stomach) of untreated female A. aegypti an increase in the surface area of the rough endoplasmic reticulum (rer) and in the ratio of membrane-bound to free ribosomes is morphometrically measured during digestion of the first blood meal. This can be correlated with the synthesis and release of digestive proteases. The dynamics of the ribosomes in A. aegypti are similar to those in A. stephensi. 3 ng alpha-amanitin per mosquito prevent normal blood digestion, the proliferation of the rer and the increase in the ratio of bound to free ribosomes. On the other hand, some synthesis of new ribosomes takes place.     

Zonal changes in the rat liver after chronic administration of phenobarbitone in ultrastructural, morphometric and biochemical correlation.

Alterations in the liver of rats subjected to 24 days of continuous administration of phenobarbitone have been supplied bu subcellular fractionation, conventional electron microscopy and morphometric analysis. The increase in wet weight of the liver was found to result from a combination of cellular hypertrophy, hyperplasia and an enlarged hepatic blood space. In the centrilobular zone all the hepatocytes underwent a substantial proliferation of total ER, became enlarged and had an increased blood supply. However, in the periportal zone phenobarbitone caused changes in only 45% of the hepatocytes, the remainder being apparently resistent or tardy. An overall dramatic increase in hepatic RER was both measured and observed but the response involved hepatocytes in which the RER had proliferated as well as those which were depleted of RER or had stacks and cisternae that were severely shortened and dispersed. These alterations are discussed in relation to changes in RER after administration of agents causing hepatonecrosis. Possible reasons for the inability of other workers to detect a phenobarbitone-induced increase in RER are also put forward. After subcellular fractionation and corection for centrifugation losses into the 9500 g pellet, using the microsomal marker cytochrome P-450, phenobarbitone-induced increase in total ER was substantially less than that found by morphometric analysis. This indicates that during the preparation of microsomes a substantial proportion of intracellular membranes, having different metabolic and synthetic properties to those finally isolated, are discarded and emphasizes the need to exercise care when using microsomal preparations.     

Ultrastructural changes in the midgut epithelium in Podura aquatica L. (Insecta, Collembola, Arthropleona) during regeneration

Data considering the degeneration and regeneration of the midgut epithelium in the primitive wingless insects, such as Collembola, are rather poor. Also information, which treats the regenerative cells as the primordial cells, is poorly known. The midgut epithelium of Podura aquatica L. (Insecta, Collembola, Arthropleona) is formed by the epithelial and regenerative cells. The epithelial cells show distinct regionalisation in the organelles distribution. The ultrastructure of the basal, perinuclear and apical regions of the epithelial cells is described. As in insects without Malpighian tubules, structures which resemble urospherites occur in the cytoplasm of the epithelial cells. After degeneration of the entire midgut epithelium, a new epithelium is formed from regenerative cells. During the process of regeneration, the degenerated epithelium gradually is separated from the basal lamina by the newly formed one. Finally, the detached epithelium is moved into the midgut lumen. Regenerative cells play a role of primordial cells during epithelial regeneration.     

Post-feeding induction of trypsin in the midgut of Aedes aegypti L. (Diptera: Culicidae) is separable into two cellular phases

The induction of trypsin activity in the midgut of the mosquito, Aedes aegypti, was studied following meals of chicken blood, and several protein and peptide diets. Various concentrations of bovine serum albumin (BSA) in 0.15 M NaCl stimulated trypsin activity, in a similar fashion to the initial increase observed after a normal blood meal. Trypsin synthesis was also initiated when Ae. aegypti were fed on glutaraldehyde cross-linked BSA and on BSA fragments prepared by both pepsin and cyanogen bromide cleavage. Non-soluble proteins, in the form of glutaraldehyde-fixed erythrocyte ghosts, induced a delayed and reduced trypsin response, whilst small peptides from neutralized liver digests did not induce trypsin activity until 8–10 h after feeding. Metabolic inhibitors had varying effects on the post-feeding activity of trypsin stimulated by BSA feeding. Cycloheximide, a peptidyl transferase inhibitor prevented expression of all activity in vivo, whereas α-amanitin (RNA-polymerase inhibitor) did not affect trypsin activity in the first 10 h after feeding. At 20 μg/ml concentration in the diet, actinomycin D (RNA synthesis inhibitor) caused temporary superinduction followed by inhibition of trypsin activity, but at lower concentrations, the later phase of trypsin activity was inhibited. The results suggest that post-feeding induction of trypsin activity in Ae. aegypti is a two-phase process regulated at the midgut cellular level. The first phase of trypsin synthesis is stimulated by soluble proteins of variable molecular weights, and only involves translation of messenger RNA already available within the midgut cells. The second phase is stimulated by small peptides and requires complete synthesis of new mRNA from DNA.     

Ultrastructural studies of midgut epithelium formation in Lepisma saccharina L. (Insecta, Zygentoma)

At the end of embryogenesis of Lepisma saccharina L. (Insecta, Zygentoma), when the stomodaeum and proctodaeum are completely formed, the midgut epithelium is replaced by the primary midgut, a yolk mass is surrounded by a cell membrane. Midgut epithelium formation begins in the 1st larval stage. Energids migrate toward the yolk periphery and aggregate just beneath the cell membrane. They are gradually enclosed by cell membrane folds of the primary midgut. Single cells are formed. Succeeding energids join just formed cells. Thus, groups of cells, regenerative cell groups, are formed. Their number gradually increases. The external cells of the regenerative cell groups transform into epithelial cells and their basal regions spread toward the next regenerative cell groups. Epithelial cells of neighboring regenerative cell groups join each other to form the epithelium. At the end of the 2nd larval stage, just before molting, degeneration of newly the formed epithelium begins. Remains of organelles and basal membrane occur between the regenerative cell groups. The new epithelium is formed from the regenerative cell groups, which are now termed stem cells of the midgut epithelium.     

Ultrastructural changes in the muscles,midgut, hemopoietic organ,imaginal discs and Malpighian tubules of the mosquito Aedes aegypti larvae infected by the fungus Coelomomyces stegomyiae

Fungi belonging to the genus Coelomomyces can infect mosquito larvae and develop within the larval hemocoel. To examine fungal development, Aedesaegypti larvae infected with Coelomomyces stegomyiae Keilin were fixed, embedded and sectioned for both light and electron microscopy. While fungal hyphae of C. stegomyiae did not invade cells other than the cuticular epithelial cells, they did penetrate a number of tissues including muscles, midgut, hemopoietic organ, imaginal discs, and Malpighian tubules. This revised version was published online in July 2006 with corrections to the Cover Date.     

Spatial and temporal patterns of abundance of Aedes aegypti L. (Stegomyia aegypti) and Aedes albopictus (Skuse) [Stegomyia albopictus (Skuse)] in southern Florida

Invasion by mosquito vectors of disease may impact the distribution of resident mosquitoes, resulting in novel patterns of vectors and concomitant risk for disease. One example of such an impact is the invasion by Aedes albopictus (Skuse) [Stegomyia albopictus (Skuse)] (Diptera: Culicidae) of North America and this species' interaction with Aedes aegypti L. (Stegomyia aegypti L). We hypothesized that Ae. aegypti would be found in urban, coastal areas that experience hotter and drier conditions, whereas Ae. albopictus would be more commonly found in suburban and rural areas that are cooler and wetter. In addition, we hypothesized that Ae. aegypti would be more abundant early in the wet season, whereas Ae. albopictus would be more abundant later in the wet season. Urban areas were drier, hotter and contained more Ae. aegypti than suburban or rural areas. Aedes aegypti was relatively more abundant early in the wet season, whereas Ae. albopictus was more abundant in both the late wet season and the dry season. The spatial patterns of inter‐ and intraspecific encounters between these species were also described. The distribution of these mosquitoes is correlated with abiotic conditions, and with temperature, humidity and the relative availability of rain‐filled containers. Understanding the ecological determinants of species distribution can provide insight into the biology of these vectors and important information for their appropriate control.     

The potential attractant or repellent effects of different water types on oviposition in Aedes aegypti L. (Dipt., Culicidae)

Abstract:  The selection of oviposition sites by the yellow-fever mosquito, Aedes aegypti , was studied in the laboratory. The repellent or attractant effects of salinity and the presence of bacteria in water collected from a local community on the Brazilian coast were investigated. Water contaminated with bacteria (waste water and lagoon water) elicited significantly more oviposition than distilled water. Oviposition decreased with an increase in salinity, with almost no oviposition above 12%. The salinity tolerance of A. aegypti larvae was investigated by measuring their mortality in response to salt concentrations ranging from 10 to 15%. The LD₅₀ of salt for A. aegypti larvae was around 13%.     

Proteomic identification of Bacillus thuringiensis subsp. israelensis toxin Cry4Ba binding proteins in midgut membranes from Aedes (Stegomyia) aegypti Linnaeus (Diptera,Culicidae) larvae

Novel Bacillus thuringiensis subsp. israelensis (Bti) Cry4Ba toxin-binding proteins have been identified in gut brush border membranes of the Aedes (Stegomyia) aegypti mosquito larvae by combining 2-dimensional gel electrophoresis (2DE) and ligand blotting followed by protein identification using mass spectrometry and database searching. Three alkaline phosphatase isoforms and aminopeptidase were identified. Other Cry4Ba binding proteins identified include the putative lipid raft proteins flotillin and prohibitin, V-ATPase B subunit and actin. These identified proteins might play important roles in mediating the toxicity of Cry4Ba due to their location in the gut brush border membrane. Cadherin-type protein was not identified, although previously, we identified a midgut cadherin AgCad1 as a putative Cry4Ba receptor in Anopheles gambiae mosquito larvae [Hua, G., Zhang, R., Abdullah, M.A., Adang, M.J., 2008. Anopheles gambiae cadherin AgCad1 binds the Cry4Ba toxin of Bacillus thuringiensis israelensis and a fragment of AgCad1 synergizes toxicity. Biochemistry 47, 5101–5110]. Other identified proteins in this study that might have lesser roles include mitochondrial proteins such as ATP synthase subunits, mitochondrial processing peptidase and porin; which are likely contaminants from mitochondria and are not brush border membrane components. Trypsin-like serine protease was also identified as a protein that binds Cry4Ba. Identification of these toxin-binding proteins will lead to a better understanding of the mode of action of this toxin in mosquito.     

Specialization of the endoplasmic reticulum in the apex of the midgut cells of Aeshna cyanea (Insecta,Odonata)

Summary Lamellar stacks of cisternae regularly spaced and bound to each other by fine filaments are observed in the apex of midgut cells of Aeshna cyanea. These cisternae are connected with the rough endoplasmic reticulum but are devoid of ribosomes except on their terminal sacs. Their location and their structure, quite different in fed and starved animals, suggest that they are involved in intracellular transport.