Targeting Plasmodium ligands on mosquito salivary glands and midgut with a phage display peptide library

Despite vast efforts and expenditures in the past few decades, malaria continues to kill millions of persons every year, and new approaches for disease control are urgently needed. To complete its life cycle in the mosquito, Plasmodium, the causative agent of malaria, has to traverse the epithelia of the midgut and salivary glands. Although strong circumstantial evidence indicates that parasite interactions with the two organs are specific, hardly any information is available about the interacting molecules. By use of a phage display library, we identified a 12-aa peptide-salivary gland and midgut peptide 1 (SM1)-that binds to the distal lobes of the salivary gland and to the luminal side of the midgut epithelium, but not to the midgut surface facing the hemolymph or to ovaries. The coincidence of the tissues with which parasites and the SM1 peptide interact suggested that the parasite and peptide recognize the same surface ligand. In support of this hypothesis, the SM1 peptide strongly inhibited Plasmodium invasion of salivary gland and midgut epithelia. These experiments suggest a new strategy for the genetic manipulation of mosquito vectorial capacity.

Distribution of acid phosphatases in the hypopharyngeal glands from workers, queens and males of a Brazilian stingless bee Scaptotrigona postica Latreille: An ultrastructural cytochemical study

The present study reports the localization of acid phosphatase in the hypopharyngeal gland cells from workers (newly-emerged, nurse and forager), queens (newly-emerged and laying) and males (newly-emerged and mature for mating) of the Brazilian stingless bee, Scaptotrigona postica. The phosphatase activity varied in intensity and localization depending on the individual class, physiological age and the substrate used. In newly-emerged workers, the phosphatase-positive sites suggest the involvement of the enzyme with cellular differentiation that occurs in the presecretory phase, in nurse workers with protein synthesis and in forager workers with changes in cellular activity or glandular regression. In males mature for mating and laying queens, the positive sites are related to secretory activity, showing that the gland maintains some activity in spite of the regressive aspect. Of the substrates used, β-glycerophosphate gave the least specific localization.

Exocrine glands of Schwarziana quadripunctata (Hymenoptera, Apinae, Meliponini)

This article describes the location, anatomy, histology and ontogeny of adult Schwarziana quadripunctata exocrine glands. These glands appear either as individualized organs (salivary gland system and Dufour gland) or as epidermis differentiation (tegumentary glands). Variations in the occurrence and degree of development among colony components with regard to their degree of maturity are also described.

Experimental control of the effect of extra doses of juvenile hormone on bee development: The case of the wax glands of Apis mellifera (Hymenoptera: Apidae)

Experiments on the effect of topical application of the synthetic juvenile hormone (JH-III) and of the solvent used to dissolve the hormone on the development of the wax glands of workers of Apis mellifera, were made. The results show that it was impossible to determine the effect of the juvenile hormone (JH) apart from its solvent (acetone), which also alters the developmental pattern of the gland. Most of the experiments reported in scientific literature do not consider the effect of the solvent, analyzing the results by only comparing the treatment with the hormone plus solvent to a control without any treatment. The data presented suggests that this kind of procedure compromises the evaluation of the real JH effect.

Histological and morphometric comparisons of worker and queen mandibular glands of Melipona bicolor bicolor (Hymenoptera, Meliponini)

In the present investigation we analyzed both histology and area of secretory cells from workers, virgin and physogastric queen mandibular glands of Melipona bicolor bicolor Lepeletier. The histological differences observed include the number of vacuoles present in the cell cytoplasm and the quantity and distribution of the secretion. The morphometry results demonstrated the occurrence of significative differences in the cell area values that were related to the secretory cell activity level, which is higher in the physogastric queens. Worker and queen glands have histological patterns quite similar, but the morphometric data indicated cast-specific differences, as well as differences between the queens, which were related to the posture activity.

Ultrastructure of the salivary glands of Pachycondyla (=Neoponera) villosa (Fabricius) (Formicidae: Ponerinae): Functional changes during the last larval instar

The cells of secretory region of the salivary glands of Pachycondyla (=Neoponera) villosa at the time of enzyme production presents the basal cellular membranes profusely folded and the intercellular junctional membranes present a few enlarged spaces. The rough endoplasmic reticulum and the Golgi bodies shift from being flat and small vesicular cisternae to enlarged vesicular cisternae according to the cell physiological state and characterize an asynchronic cell cycle. Enzymes are released into the lumen by microapocrine secretion. The stage of silk production is detected after a behavioral act, when the nurse worker separates the mature larva. At this time, the salivary gland cells present only one physiological state (synchronized secretory cycle): this state was characterized by basal cellular membrane poorly folded, intercellular junctions presenting some small spaces, rough endoplasmic reticulum compounded by flat cistenae, enlarged Golgi bodies with fibrous material inside and a few secretory vesicles containing silk, which undergo exocytosis. The silk in the lumen shows 2 forms: tactoid and flocculent material.

Little cicada of sugarcane Mahanarva posticata (Homoptera: Cercopidae). A Brazilian agricultural pest. Morpho-histological study of salivary glands

Little cicadas are homopteran insect pests of sugarcane plantations. As these insects suck out the sap from the leaf parenchyma, they inoculate a toxic saliva that damages the plant vessels, thus promoting the loss of glucose by the affected plant. The morphological and histological analyses of the salivary glands of the little cicada Mahanarva posticata, revealed that these glands are formed by 2 portions: one portion comprises a group of acini and has been denominated as the principal gland; the second portion is filamentous in nature and has been denominated as the accessory gland; it is formed by very long and fine filaments. The acinous portion of the gland can be subdivided into 2 lobes: an anterior lobe formed by 3 lobules (I, II, III), and a posterior lobe formed by lobule IV and the excretory duct. Histologically, the salivary glands showed that the filaments are empty sutructures composed by several internal channels with secretion granules being observed in the cytoplasm of the cells of the secretory filaments. Lobules I and II of the principal gland are characterized by being highly basophilic and for accumulating a large amount of secretion in both the cytoplasm of the cells and inside secretion vesicles. Histochemically, we verified that the secretion produced by these glands is lipidic and protein in nature, with the production of polysaccharides being very low. The differences in stain and appearance of the different regions of the salivary gland lead us to believe that the final glandular product is lipoproteic in nature.

Morphological aspects of Culex quinquefasciatus salivary glands

The salivary glands of Culex quinquefasciatus female mosquitoes are paired organs composed of two lateral lobes with proximal and distal secretory portions, and a medial lobe. All portions comprise a simple epithelium that surrounds a salivary duct. In the apical portion of the medial lobe, non-secretory cells strongly resemble cells involved in ion and water transport. The general architecture of the secretory portions is similar between lobes. The appearance of the secretory material and the morphological aspect of the apical cell membrane are the most distinctive features among the three secretory portions. Cells in the lateral proximal lobe display thin membrane projections extending into a translucent and finely filamentous secretory product. At the lateral distal portion, the apical cell membrane forms an intricate meshwork that encloses a dark secretory product. Medial lobe secretory cells also contain secretory cavities surrounded by intracytoplasmic vesicles, all containing a very dark and uniform product. Scattered cells holding numerous vacuoles, some of them containing a small and electron-dense granule eccentrically located and resembling those of the diffuse endocrine system, are frequently observed in the periphery of all secretory portions. Immunofluorescence assays revealed that the distal portion of the lateral lobes contains apyrase, an enzyme putatively responsible for platelet aggregation inhibition, diffusely distributed in the cell cytoplasm. © 2003 Elsevier Ltd. All rights reserved.

Electrophoretical Protein Profile in Apis mellifera (Hymenoptera; Apidae) Hypopharyngeal Glands under Juvenile Hormone Effect When the Colony is Grown in the Absence of Brood

Twelve-day-old and 25-day-old Apis mellifera workers were treated or not treated with juvenile hormone at the moment of emergence and reared in the colony without brood. Having the brood interference apart, the hormone effect on the hypopharyngeal glands protein expression was determined through the electrophoretical protein profiles of the both groups of bees. In those conditions, the hormone induced changes that were different from the control. Protein bands of 66 and 48 kDa were intensified in the 12-day-old bees, whereas band of 42 kDa was reduced in the 25-day-old bees. That indicated a different effect of the juvenile hormone in the function of bee aging, which promoted a glandular protein activation in the young bees and, in contrast, an inhibitory action in the 25-day-old bees workers.

Histochemistry and protein profile of the mandibular glands of workers of the ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

Insect mandibular glands are always associated to the mandibles; they are part of the salivary glandular system. The mandibular glands are composed by a reservoir associated to the secretory cells, with each secretory cell connected to the reservoir by means of individual canaliculi. These glands play an important role in the production of pheromones, which are compounds involved in defense, communication, and reproduction of the colony. Mandibular glands of soldiers and major and minor workers of the ant Atta sexdens rubropilosa were processed for different histochemical tests, total protein content, and protein electrophoretic profile determination. The histochemical tests detected the presence of lipids, DNA/RNA, polysaccharides, and proteins at different regions of the gland. The protein electrophoretic profiles showed that the total protein content as well as the number of peptides of each caste follow a progressive order in relation to the size of the individual. Thus, we suggest that the production of secretion is directly linked to the task that the individual performs in the colony.

A new interpretation of the defense glands of neotropical Ruptitermes (Isoptera, Termitidae, Apicotermitinae)

Suicidal altruism has been reported for some species of eusocial insects, in which the individual dies in defense of the society. The termites of the genus Ruptitermes are known for the suicidal behavior of the workers which liberate a sticky defensive secretion by body bursting. In the present paper it is given a new interpretation of the defense glands of Neotropical Ruptitermes based on the morphological analysis of three species collected at Rio Claro, SP, Brazil. Before the current study, the suicidal defensive behavior was attributed to the dehiscence of the salivary gland reservoirs. The defense or dehiscent glands of Neotropical Ruptitermes are pair structures rounded in shape that are independent of the salivary glands. The dehiscent glands consist of multiple secretory units that are kept together by thin connective tissue. Each secretory unit is composed of one cell generally with one peripheral nucleus and characteristic secretion. The three species studied here present some histological differences in the secretory units, probably related to the chemical composition of the secretion.

Study of the venom glands in Ectatomma quadridens (Hymenoptera: Formicidae). Evolutionary hypothesis in subfamily ponerinae

The venom glands of worker ants of the species Ectatomma quadridens morphologically resemble an elongated sac or reservoir ending in a narrower portion that has the function of releasing the secretion to the exterior. Two external secretory filaments are individually inserted into the proximal portion of the gland and end inside the convoluted gland. The venom gland of workers of E. quadridens is, therefore, morphologically subdivided into four distinct portions: a) sac-shaped reservoir measuring approximately 1mm in length; b) excretory duct, proximal portion of the reservoir that joins the gland to the sting apparatus; c) convoluted gland, final portion of the external secretory filaments located inside the reservoir; and d) two secretory filaments measuring about 2 mm in length; their free extremities end blindly and are individually inserted into the reservoir wall at the proximal region of the venom gland. The histological data showed that the filaments and the convoluted gland are composed of cubic cells of secretory function. The reservoir consists of a simple cubical epithelium externally surrounded by muscle fibers. A thick cuticle internally coats the epithelium of the reservoir. The application of histochemical tests allowed us to establish that the final secretion of the venom gland of Ectatomma quadridens is of glycoproteic nature. This secretion undergoes several modifications at the secretory filaments, at the convoluted gland, and in the reservoir before reaching the excretory duct, the point at which the secretion is released in its final composition, namely the venom. Based on the differences among various Ponerinae species we propose a hypothesis suggesting a probable evolutionary process that the venom glands of members of this subfamily might have undergone.

Morphology and protein patterns of honey bee drone accessory glands.

We used light and transmission electron microscopy to examine the morphology of the accessory glands of immature and mature adult males of Apis mellifera L. We also made an electrophoretic analysis of the protein content of the mature gland. The glands of the immature male actively secrete a mucous substance that can be seen in the lumen of the gland of the mature male. This secretion stains with mercury bromophenol blue and with periodic acid-Schiff reaction, which stain glyconjugates. The protein content was higher in the lumen secretion than in the gland wall extracts. The electrophoresis patterns of the wall extracts were different from those of the secretion found in the gland lumen.

Hydrolases in the hypopharyngeal glands of workers of Scaptotrigona postica and Apis mellifera (Hymenoptera, Apinae)

Hydrolytic enzymes from hypopharyngeal gland extracts of newly emerged, nurse and foraging workers of two eusocial bees, Scaptotrigona postica, a native Brazilian stingless bee, and the Africanized honey bee (Apis mellifera) in Brazil, were compared. The hypopharyngeal gland is rich in enzymes in both species. Fifteen different enzymes were found in the extracts, with only a few quantitative differences between the species. Some of the enzymes present in the extracts may have intracellular functions, while others seem to be digestive enzymes. Scaptotrigona postica, had lower β-glucosidase and higher lipase esterase activities than A. mellifera. The differences may be due to different feeding habits and behavioral peculiarities of the two species. ©FUNPEC-RP.