Ultrastructural changes in the abdominal midgut of the mosquito, Culiseta melanura, during the gonotrophic cycle

Abdominal midguts of the mosquito, Culiseta melanura, were examined by light and electron microscopy 1 hr-14 days days after blood feeding. Epithelial cells were drastically altered from columnar to squamous in form after engorgement, and returned to columnar by day 4 after feeding. Accumulation of mitochondria along brush borders of digestive cells, followed by the appearance of large secondary lysosomes, accompanied blood digestion. Evidence was obtained that myelin-like material in the lysosomes, probably the result of mitochondrial autolysis, is extruded into the lumen. Digestive cells resumed their pre-blood meal appearance by 10-14 days post-engorgement. Regenerative cells were scattered throughout the basal portion of the epithelium, along with endocrine cells. Other midgut cells containing large, microvilli-lined apical cavities were identified in most specimens. No evidence of division or differentiation was obtained for any cell types.     

Ovary ecdysteroidogenic hormone functions independently of the insulin receptor in the yellow fever mosquito,Aedes aegypti

Most mosquito species must feed on the blood of a vertebrate host to produce eggs. In the yellow fever mosquito, Aedes aegypti, blood feeding triggers medial neurosecretory cells in the brain to release insulin-like peptides (ILPs) and ovary ecdysteroidogenic hormone (OEH). Theses hormones thereafter directly induce the ovaries to produce ecdysteroid hormone (ECD), which activates the synthesis of yolk proteins in the fat body for uptake by oocytes. ILP3 stimulates ECD production by binding to the mosquito insulin receptor (MIR). In contrast, little is known about the mode of action of OEH, which is a member of a neuropeptide family called neuroparsin. Here we report that OEH is the only neuroparsin family member present in the Ae. aegypti genome and that other mosquitoes also encode only one neuroparsin gene. Immunoblotting experiments suggested that the full-length form of the peptide, which we call long OEH (lOEH), is processed into short OEH (sOEH). The importance of processing, however, remained unclear because a recombinant form of lOEH (rlOEH) and synthetic sOEH exhibited very similar biological activity. A series of experiments indicated that neither rlOEH nor sOEH bound to ILP3 or the MIR. Signaling studies further showed that ILP3 activated the MIR but rlOEH did not, yet both neuropeptides activated Akt, which is a marker for insulin pathway signaling. Our results also indicated that activation of TOR signaling in the ovaries required co-stimulation by amino acids and either ILP3 or rlOEH. Overall, we conclude that OEH activates the insulin signaling pathway independently of the MIR, and that insulin and TOR signaling in the ovaries is coupled.     

Ultrastructural and immunohistochemical study of endocrine cells in the midgut of the cockroach Blaberus craniifer (Insecta,Dictyoptera)

Summary The midgut of Blaberus craniifer is principally made up of columnar epithelial cells which are derived from small regenerative cells found grouped in nidi. Between them, small sparsely granulated cells with clear cytoplasm can be observed lying on the basal lamina. Mainly based on the size, shape and texture of their secretory granules, at least ten types of such endocrine cells have been identified. Five cell types contain a uniform population of dense granules: (1) medium-sized, round to oval granules; (2) small elongated granules; (3) large irregular granules; (4) oval granules with a highly osmiophilic core; (5) oval, haloed granules. Five others are characterized by a heterogeneous population of granules: (6) small, round to oval, variably electron-dense granules; (7) oval medium-sized granules of variable electron density; (8) large irregular granules of variable electron density; (9) small dense granules and large vesicles with filamentous material; (10) small dense granules and very large pale vesicles.In addition, near the regenerative cells, large cells characterized by very large, irregular, dense granules (up to 4 m), lack contact with the lumen, and reach the basal lamina only by slender cytoplasmic processes.Several antisera raised against mammalian peptides and amine were used to reveal axonal fibers and endocrine cells. Serotonin-like immunoreactivity is localized in a profuse innervation of the muscle layers that surround the epithelium, whereas cholecystokinin and methionine-enkephalin antisera stain a more moderate number of axonal fibers. Cholecystokinin-, methionine-enkephalin-, substance P-, vasoactive intestinal peptide-, somatoliberin-, and gonadoliberin-like immunoreactivities were detected in endocrine cells of the epithelium. While most of the cells appear pyramidal, oval, fusiform or bowl-shaped, and seem to lack contact with the lumen, cells reaching it have been detected reacting with antisera to cholecystokinin, substance P, vasoactive intestinal peptide, somatoliberin and gonadoliberin.     

The fine structural morphology of the midgut of adult Drosophila: A morphometric analysis

The midgut of one day old Drosophilia was examined morphometrically at the electron microscopic level. Results suggest that parenchymal cells, with the exception of basal cells, possess identical functions. Drosophilia midgut cells are smaller than those of other insects studied, and the surface densities of the rER was less, indicating that its protein synthetic activity is also less than that of other insects.     

Ultrastructure and functional features of midgut of an adult water mite Teutonia cometes (Koch, 1837) (Hydrachnidia: Teutoniidae)

Shatrov, A. B. 2010. Ultrastructure and functional features of midgut of an adult water mite Teutonia cometes (Koch 1837) (Hydrachnidia: Teutoniidae). —Acta Zoologica (Stockholm) 91 : 222–232 The midgut of the adult water mite Teutonia cometes (Koch 1837) (Hydrachnidia: Teutoniidae) was investigated by means of transmission electron microscopy and on semi‐thin sections. The midgut is represented by a blind sac composed of the narrow ventriculus, two proventricular lateral diverticula and three pairs of postventricular caeca. A single‐layered epithelium consists of one type of endodermal digestive cells of quite different shape and size, which may form protrusions into the midgut lumen. The large nuclei are frequently lobed and contain one to three nucleoli. The apical cell membrane forms short scarce microvilli, between their bases the pinocytotic vesicles of unspecific macropinocytosis as well as the narrow pinocytotic canals are formed and immersed into the cell. The intracellular digestion of the food ingested into the midgut after extraintestinal digestion is predominant. The pinocytotic vesicles fuse with small clear vesicles of proposed Golgi origin to form secondary lysosomes. The digestive cells also contain small amounts of rough endoplasmic reticulum, variously structured heterolysosomes, residual materials in the form of both the small electron‐dense bodies and the large variously granulated substances, reserve nutritive materials such as lipid and glycogen, as well as clear vacuoles. Residual materials are obviously extruded from the cells into the gut lumen.     

Imidacloprid impairs the post‐embryonic development of the midgut in the yellow fever mosquito Stegomyia aegypti (=Aedes aegypti)

The mosquito Stegomyia aegypti (=Aedes aegypti) (Diptera: Culicidae) is a vector for the dengue and yellow fever viruses. As blood digestion occurs in the midgut, this organ constitutes the route of entry of many pathogens. The effects of the insecticide imidacloprid on the survival of St. aegypti were investigated and the sub‐lethal effects of the insecticide on midgut development were determined. Third instar larvae were exposed to different concentrations of imidacloprid (0.15, 1.5, 3.0, 6.0 and 15.0 p.p.m.) and survival was monitored every 24 h for 10 days. Midguts from imidacloprid‐treated insects at different stages of development were dissected and processed for analyses by transmission electron microscopy, immunofluorescence microscopy and terminal deoxynucleotidyl transferase dUTP nick‐end labelling (TUNEL) assays. Imidacloprid concentrations of 3.0 and 15.0 p.p.m. were found to affect midgut development similarly. Digestive cells of the fourth instar larvae (L4) midgut exposed to imidacloprid had more multilamellar bodies, abundantly found in the cell apex, and more electron‐lucent vacuoles in the basal region compared with those from untreated insects. Moreover, imidacloprid interfered with the differentiation of regenerative cells, dramatically reducing the number of digestive and endocrine cells and leading to malformation of the midgut epithelium in adults. The data demonstrate that imidacloprid can reduce the survival of mosquitoes and thus indicate its potentially high efficacy in the control of St. aegypti populations.     

Biochemical screening of Lactobacilli in midgut of wild Aedes aegypti mosquitoes. A step toward paratransgenesis candidate

This study was conducted to isolate and identify lactobacilli from larval and adult midgut of wild Aedes aegypti (Ae. aegypti) to find candidate bacteria for paratransgenic control. Characterization of the bacterial symbionts was done using Gram staining, motility test, catalase test, and biochemical tests, among others, and the morphological features were confirmed using a standard scheme that simplifies the identification of lactic acid bacteria. A total of 174 strains were isolated and identified, 135 strains from larval midgut and 39 strains from adult midgut (mean ± SE, 4.00 ± 0.72; P = 0.00). The isolated species were confirmed to be Lactobacillus fermentum, L. casei, L. acidophilus, L. viridescens, L. brevis and L. gasseri. It can be concluded that Ae. aegypti has the potential of harboring the cultivable bacterial symbionts. In conclusion, the isolated species were nominated for paratransgenic control, particularly L. fermentum, being found in large numbers from both larval and adulxt midgut.     

Intestinal stem cells in the adult Drosophila midgut

Drosophila has long been an excellent model organism for studying stem cell biology. Notably, studies of Drosophila's germline stem cells have been instrumental in developing the stem cell niche concept. The recent discovery of somatic stem cells in adult Drosophila, particularly the intestinal stem cells (ISCs) of the midgut, has established Drosophila as an exciting model to study stem cell-mediated adult tissue homeostasis and regeneration. Here, we review the major signaling pathways that regulate the self-renewal, proliferation and differentiation of Drosophila ISCs, discussing how this regulation maintains midgut homeostasis and mediates regeneration of the intestinal epithelium after injury.     

Ultrastructure and morphology of midgut visceral muscle in early pupal Aedes aegypti mosquitoes

These studies focus on the pupal Aedes aegypti midgut muscularis for the first 26 h following larval-pupal transition. The midgut muscularis of Ae. aegypti pupae during this first half of the pupal stadium is a grid of both circularly and longitudinally oriented muscle bands, arranged in a manner resembling that of the larvae. While many muscle bands exhibit signs of degeneration during the time period studied, not all bands degrade, nor is this degradation simultaneous. Band deterioration involves destruction of internal elements while the muscle fiber plasma membrane remains intact. Deterioration of contractile elements may involve proteosome-like structures and associated enzymes. Many features of the larval muscularis including cruciform cells, bifurcating circular bands, and bifurcating longitudinal bands of muscle are retained during the time period investigated. Neuromuscular junctions along some muscle bands are retained through at least 16 h into the pupal stadium. The selective nature of muscle fiber degradation, coupled with the retention of larval features and neural input, may allow for limited functionality of the muscularis during metamorphosis. Evidence of sexual dimorphism in the midgut muscularis of male and female Ae. aegypti pupae was not observed during the time period studied.     

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.     

Ultrastructure and immunocytochemistry of endocrine cells in the midgut of the desert locust,Schistocerca gregaria (Forskal)

Summary The endocrine cells of the midgut epithelium of the desert locust are found dispersed among the digestive cells and are similar to those of the vertebrate gut. According to their reactivity to silver impregnation techniques and the ultrastructural features of the secretory granules (shape, electron-density, size, and structure) 10 types of endocrine cell have been identified, of which seven are located in the main segment of the midgut or in the enteric caeca, and the other three seem to be present only in the ampullae through which the Malpighian tubules drain into the gut. The endocrine cells have a slender cytoplasmic process that reaches the gut lumen, a feature that supports the receptosecretory nature postulated for this cellular type in insects as well as vertebrates. Antisera directed against mammalian gastrin, CCK, insulin, pancreatic polypeptide and bombesin reacted with some of the endocrine cells. This is the first time that insulin- and bombesin-like immunoreactive cells have been described in the midgut of an insect.     

Distention-mediated egg maturation in the mosquito,Aedes aegypti

Abdominal distention accelerates the release of a factor from the head of blood-fed Aedes aegypti mosquitoes. The critical period during which the head is required for oögenesis following blood ingestion is approx 6 h with a 5 μl meal, but small blood meals of 1 μl require the head to be present for significantly longer. Increasing the abdominal distention by supplementing the 1 μl meal with saline results in a critical period similar to that with 5 μl of blood. The information from the distended abdomen appears to travel via the ventral nerve cord. Transection of the ventral nerve cord prevents oögenesis from occurring after small blood meals, but not with larger blood volumes. Topical application of 100 pg of juvenile hormone III can substitute for the distention message.     

Ultrastructural study of cholecystokinin-like immunoreactivity in endocrine cells of the insect midgut

Summary Electron-microscopic immunocytochemistry for the demonstration of CCK-like material in basal endocrine cells of the midgut of Aeschna cyanea, Locusta migratoria, Carausius morosus and Periplaneta americana was performed by use of the peroxidase-antiperoxidase procedure and the colloidal gold method. Immunoreactive cells appeared scattered among digestive and regenerative cells of the epithelium. Immunoreactivity was specifically detected over round to oval electron-dense granules whose size appeared rather different from species to species. Thus, the average size of 30% (d30) of the largest granules ranged from 312 nm in Periplaneta americana to 159 nm in Carausius morosus with intermediate values in Aeschna cyanea (d30=195 nm) and Locusta migratoria (d30=225 nm).     

FLP-mediated recombination in the vector mosquito, Aedes aegypti.

The activity of a yeast recombinase, FLP, on specific target DNA sequences, FRT, has been demonstrated in embryos of the vector mosquito, Aedes aegypti. In a series of experiments, plasmids containing the FLP recombinase under control of a heterologous heat-shock gene promoter were co-injected with target plasmids containing FRT sites into preblastoderm stage mosquito embryos. FLP-mediated recombination was detected between (i) tandem repeats of FRT sites leading to the excision of specific DNA sequences and (ii) FRT sites located on separate plasmids resulting in the formation of heterodimeric or higher order multimeric plasmids. In addition to FRT sites originally isolated from the yeast 2 microns plasmid, a number of synthetic FRT sites were also used. The synthetic sites were fully functional as target sites for recombination and gave results similar to those derived from the yeast 2 microns plasmid. This successful demonstration of yeast FLP recombinase activity in the mosquito embryo suggests a possible future application of this system in establishing transformed lines of mosquitoes for use in vector control strategies and basic studies.     

Toward a description of the sialome of the adult female mosquito Aedes aegypti

To describe the set of mRNA and protein expressed in the salivary glands (sialome) of Aedes aegypti mosquitoes, we randomly sequenced a full-length cDNA library of this insect and performed Edman degradation of PVDF-transferred protein bands from salivary homogenates. We found 238 cDNA clusters which contained those coding for 10 of the 11 proteins found by aminoterminal degradation. All six previously described salivary proteins were found in this library. Full-length sequences of 32 novel cDNA sequences are reported, one of which is the product of a transposable element. Among the 31 novel protein sequences are 4 additional members of the D7 protein family; 4 novel members of the antigen 5 family (a protein family not reported in Aedes); a novel serpin; a novel member of the 30-kDa allergen of Ae. Aegypti; a secreted calreticulin; 2 proteins similar to mammalian angiopoietins; adenosine deaminase; purine hydrolase; lysozyme; a C-type lectin; 3 serine proteases, including one with high similarity to Bombyx prophenoloxidase activating enzyme; 2 proteins related to invertebrate immunity; and several sequences that have no significant matches to known proteins. The possible role of these proteins in blood and sugar feeding by the mosquito is discussed.     

Regenerative cells and the architecture of beetle midgut epithelia

The architectural ground plan of beetle and other insect midguts is represented by a monolayer of epithelial cells arranged in a cylindrical configuration. Proliferation and differentiation of regenerative cells maintain the integrity of this monolayer in the face of continual losses of individual cells through cytoplasmic budding and/or expulsion of entire epithelial cells. Peritrophic membranes have conventionally been considered universal features of insect midguts that function to protect vulnerable microvillar surfaces of the midgut epithelium from abrasion by ingested food; however, peritrophic membranes were found in only a small fraction of the adult beetle species examined in this study. In adult beetles, midgut epithelial cells are continually replaced by cells recruited from populations of mitotic regenerative cells that are interspersed among the differentiated epithelial monolayer. To remain contiguous with the other cells in the midgut monolayer, some of these proliferating populations have adopted evaginated configurations of cells that extend for varying distances from the basal surface of the monolayer. These configurations are referred to as regenerative crypts or pouches and consist of progenitor cells and stem cells. The presence, the relative densities, and the relative lengths of these regenerative pouches vary considerably among families of beetles. Placement of longitudinal muscles of the midgut relative to the proximodistal axes of these regenerative pouches also varies among species of beetles. The presence, the size, and the density of regenerative cell populations are related to 1) feeding habits of adult beetles, 2) presence of peritrophic membranes, and 3) expulsion of entire midgut epithelial cells or fragments of these epithelial cells into midgut lumens. © 2012 Wiley Periodicals, Inc.