Alteration of Ca(2+)-ATPase activity in the homogenate, plasma membrane and microsomes of the salivary glands of streptozotocin-induced diabetic rats

Diabetes has been implicated in the dryness of the mouth, loss of taste sensation, sialosis, and other disorders of the oral cavity, by impairment of the salivary glands. The aim of the present study was to examine the plasma membrane, microsomal, and homogenate Ca(2+)-ATPase activity in the rat submandibular and parotid salivary glands of streptozotocin-induced diabetes. We have also examined the influence of the acidosis state oil this parameter. Diabetes was induced by an intraperitoneal injection of streptozotocin and acidosis was induced by daily injection of NH(4)Cl. At 15 and 30 days after diabetes induction, the animals were euthanized and the submandibular and parotid salivary glands were removed and analyzed. Ca(2+)-ATPase (total, independent, and dependent) was determined in the homo-enate, microsomal, and plasma membranes of the salivary glands of diabetic and control rats. Calcium concentration was also determined in the glands and showed to be hi-her in the diabetic animals. Ca(2+)-ATPase activity was found to be reduced in all cell fractions studied in the diabetic animals compared with control. Similar results were obtained for the submandibular salivary glands of acidotic animals; however in the parotid salivary glands it was found an increase in the enzyme activity. Copyright (c) 2009 John Wiley & Sons, Ltd.

Light microscopy and ultrastructure of the liver of Astyanax altiparanae Garutti and Britski, 2000 (Teleostei, Characidae).

Livers of thirty specimens of Astyanax altiparanae obtained from a commercial fish farm were subjected to light and transmission electron microscopy, in order to describe the hepatic parenchyma and the intrahepatic exocrine pancreatic tissue. Anatomically, the liver showed only three hepatic lobes. Histological analysis demonstrated that the hepatocytes were spread out as anastomotic cords, arranged in two cellular layers and surrounded by sinusoids. The intrahepatic exocrine pancreatic tissue exhibited an acinar arrangement and was diffused in the hepatic parenchyma. Ultrastructural analysis showed that the hepatocytes had a rounded nucleus and a rough endoplasmatic reticulum, with a parallel disposition to the nuclear membrane. The exocrine pancreatic cells showed secretion granules at the apical portion, and the rough endosplasmatic reticulum was concentrically distributed.

Structural and cytochemical changes in the salivary glands of the Rhipicephalus (Boophilus) microplus (CANESTRINI, 1887) (Acari : Ixodidae) tick female during feeding

This study describes the morphology of salivary glands of Rhipicephalus (Boophilus) microplus female ticks at beginning of feeding (24-48 h of attachment) and semi-engorged (4-5 days of attachment) to verify the degenerative characteristics of these organs and the secretory phase in which the process begins. At the beginning of feeding, secretion granules had been observed only in the cytoplasm of cells b, c(1), c(2), c(4) (type II acinus) and d (type III acinus), as well as large nuclei with regular and preserved morphology. In the semi-engorged females the acini presented few normal cells, few partially preserved ones, and the remaining ones in several stages of degeneration, that is, with retraction and cytoplasmic vacuolization, and nuclei with chromatin in several stages of condensation, picnotic and/or in fragmentation. In type I acinus and in the excretory ducts of the studied glands, at both feeding stages, no degenerative characteristic was observed. In females of R. (B.) microplus, the salivary glands degenerate asynchronically and precociously when compared with those of others tick's species. (c) 2006 Elsevier B.V. All rights reserved.

Ultrastructural studies of the mandibular glands of the minima, media and soldier ants of Atta sexdens rubropilosa (Forel 1908)

The mandibular glands of Hymenoptera are structures associated with the mandibles and constitute part of the salivary glands system. Histological studies in workers of Atta sexdens rubropilosa revealed that this gland contains two portions: a secretory and a storage portion or reservoir. Both portions are connected by means of canaliculi. The object of the present work was the study of the ultratructure of the mandibular glands of minima, media and soldier ant of A. s. rubropilosa by TEM techniques. The glands, in the three castes studied, possess a reservoir, constituted by a simple pavementous epithelium surrounded by the cuticular intima and the secretory portion is constituted by cells of rounded shape. The secretory cells, mainly of minima and soldier, were rich in smooth endoplasmic reticulum. The media worker and soldier presented a large number of mitochondria, of varying shape. Well-developed Golgi complexes were also present in the soldiers. The secretory cells in minima, media and soldier were provided with collecting intracellular canaliculi, which were linked to the reservoir through the extracellular portion. The cytoplasm of the canaliculi-forming cell was poor in organelles. In the individuals of the three castes of A. s. rubropilosa, the presence of lipid secretion granules suggested, beyond the other functions, also a possible pheromonal action. The different roles executed by the different insect castes are directly dependent on the glandular products and, consequently, on the secretory cellular characteristics. (c) 2005 Elsevier Ltd. All rights reserved.

Ocorrência, morfologia e ultra-estrutura da glândula de Dufour de Scaptotrigona postica Latreille (Hymenoptera: Apidae)

A glândula de Dufour é uma glândula acessória do aparelho reprodutivo feminino das abelhas. Nas abelhas neotropicais sem ferrão, tem sido pouca estudada sob todos os aspectos: morfológico, ontogenético e bioquímico. Na tentativa de colaborar com o conhecimento dessa glândula em abelhas sem ferrão, foi realizado um estudo da sua ocorrência, morfologia e desenvolvimento em Scaptotrigona postica Latreille. Os resultados mostraram que ela se encontra ausente nas operárias, como ocorre em muitas outras espécies desse grupo. Nas rainhas, as células glandulares parecem mais ativas nas virgens, possuindo uma desenvolvida rede de retículo endoplasmático liso tubular, grânulos de secreção e polirribossomos dispersos no citoplasma, além de apresentarem núcleos maiores do que os das células glandulares das fisogástricas. Nas rainhas fisogástricas há dois tipos de células glandulares, ambas aparentemente inativas sinteticamente. As glândulas das rainhas fisogástricas são claramente capazes de captar substâncias da hemolinfa, provavelmente lipídios, que não penetram nas células, mas passam pelos espaços intercelulares e, através da cutícula, chegam diretamente à luz da glândula. A bem desenvolvida dupla camada de lâmina basal ao redor da glândula pode atuar no processo de captação de substâncias da hemolinfa. A secreção, e conseqüentemente sua função, pode ser diferente nas duas classes de rainhas.

Salivary glands of Amblyomma cajennense (Acari: Ixodidae): a histological and an ultrastructural overview

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Morpho-histochemical characterization of salivary gland cells of males of the tick Rhipicephalus sanguineus (Acari: Ixodidae) at different feeding stages: description of new cell types

This study describes the changes undergone by cells of the salivary glands of unfed and feeding (at day two and four post-attachment) Rhipicephalus sanguineus males, as well as new cell types. In unfed males, types I and II acini are observed with cells undifferentiated, undefined 1 and 2 (the latter, with atypical granules), a, c1 and c3; type III is composed of cells d and e; and type IV present cells g. In males at day two post-attachment, type I acini exhibit the same morphology of unfed individuals. An increase in size is observed in types II, III, and IV, as cells are filled with secretion granules. Some granules are still undergoing maturation. In type II acinus, cells a, b and c1-c8 are observed. Cells c7 and c8 are described for the first time. Cells c7 are termed as such due to the addition of polysaccharides in the composition of the secretion granules (in unfed individuals, they are termed undefined 1). Type III acini exhibit cells d and e completely filled with granules, and in type IV, cells g contain granules in several stages of maturation. In males at day four post-attachment, type I acini do not exhibit changes. Granular acini exhibit cells with fewer secretion granules, which are already mature. In type II acini, cells a, b, c1-c5 are present, type III exhibit cells d and e, and type IV contain cells g with little or no secretion. This study shows that in the salivary glands of R. sanguineus males, cells a, c1, and c3 of type II acinus, and cells d and e of type III do not exhibit changes in granular content, remaining continuously active during the entire feeding period. This indicates that during the intervals among feeding stages, gland cells reacquire the same characteristics found in unfed individuals, suggesting that they undergo reprogramming to be active in the next cycle.

Ultrastructural profile of metapleural gland cells of the ant Atta laevigata (F. Smith, 1858) (Formicidae: Attini)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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.

Rhipicephalus (Boophilus) microplus (Canestrini, 1887) (Acari: Ixodidae): Acid phosphatase and ATPase activities localization in salivary glands of females during the feeding period

This study investigates the presence and the localization of acid phosphatase and ATPase in the salivary glands of Rhipicephalus (Boophilus) microplus female ticks during feeding. Semi-engorged females showed a larger amount of acid phosphatase compared to those at beginning of feeding, localized mainly in the apical portion of the secretory cells, and in the basal labyrinth of the interstitial cells. Ultrastructural observations also demonstrated its presence in secretion granules and inside some nuclei of secretory cells at beginning of feeding. Acid phosphatase in a free form probably has a hemolymph and/or ribosomal origin and participates in salivary gland secretion control. ATPase was detected in basal membrane of all types of acini and/or in the cytoplasm of the secretory cells at both feeding stages. The enzyme activities found strongly suggests that cell death by apoptosis occurs during the degenerative process. © 2006 Elsevier Inc. All rights reserved.

Estudos morfológicos e histoquímicos do tubo digestivo de Hemisorubim platyrhynchos (Valenciennes, 1840)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

A glândula metapleural e suas secreções em formigas cultivadoras (Attini) e não cultivadoras de fungos (Myrmicini, Blepharidattini e Ectatommini) (Hymenoptera: Formicidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ultrastructure of the parotid and submandibular glands of the Old World marten (Carnivora; Mustelidae)

The morphology of the parotid and submandibular glands in the marten, a carnivore, were studied and analyzed under a transmission electron microscope. The nature of the granules in both glands, as well as in the acini and in the secretory tubules, is rather mucous. The structure of the secretory tubules is very characteristic, especially the striated ones. The myoepithelial cells are close to the acini and tubules and covered by the basement membrane separating them from the connective tissue, which enhances its epithelial origin. The cytoplasm of the basal parts of the acinar and tubular cells is abundant and separates the nucleus from the secretion granules. Although the morphology of the salivary glands of many carnivores is known, those of the parotid gland of the marten present peculiar characteristics, since they produce a rather mucous saliva and the granules, when forming, are far from the base as well as from the apex of the secretory cells. The submandibular gland contains granules of different densities, an aspect that in general resembles that of other animals.

Insulin secretion is regulated by the glucose-dependent production of islet beta cell macrophage migration inhibitory factor.

Macrophage migration inhibitory factor (MIF), originally identified as a cytokine secreted by T lymphocytes, was found recently to be both a pituitary hormone and a mediator released by immune cells in response to glucocorticoid stimulation. We report here that the insulin-secreting beta cell of the islets of Langerhans expresses MIF and that its production is regulated by glucose in a time- and concentration-dependent manner. MIF and insulin colocalize by immunocytochemistry within the secretory granules of the pancreatic islet beta cells, and once released, MIF appears to regulate insulin release in an autocrine fashion. In perifusion studies performed with isolated rat islets, immunoneutralization of MIF reduced the first and second phase of the glucose-induced insulin secretion response by 39% and 31%, respectively. Conversely, exogenously added recombinant MIF was found to potentiate insulin release. Constitutive expression of MIF antisense RNA in the insulin-secreting INS-1 cell line inhibited MIF protein synthesis and decreased significantly glucose-induced insulin release. MIF is therefore a glucose-dependent, islet cell product that regulates insulin secretion in a positive manner and may play an important role in carbohydrate metabolism.

Defining secretion processes in astrocytes

During the last decade, evidence that release of chemical transmitters from astrocytes might modulate neuronal activity (the so-called "gliotransmission") occurs in situ has been extensively provided. Nevertheless, gliotransmission remains a highly debated topic because of the lack of direct morphological and functional evidence. Here we provided new information supporting gliotransmission, by i) deepen knowledge about specific properties of regulated secretion of glutamatergic SLMVs, and ii) investigating the involvement of astrocytes in the transmission of dopamine, a molecule whose interaction with astrocytes is likely to occur, but it's still not proven.¦VGLUT-expressing glutamatergic SLMVs have been previously identified both in situ and in vitro, but description of kinetics of release were still lacking. To elucidate this issue, we took advantage of fluorescent tools (styryl dyes and pHluorin) and adapted experimental paradigms and analysis methods previously developed to study exo-endocytosis and recycling of glutamatergic vesicles at synapses. Parallel use of EPIfluorescence and total internal reflection (TIRF) imaging allowed us to find that exo-endocytosis processes in astrocytes are extremely fast, with kinetics in the order of milliseconds, able to sustain and follow neuronal signalling at synapses. Also, exocytosis of SLMVs is under the control of fast, localized Ca2+ elevations in close proximity of SLMVs and endoplasmatic reticulum (ER) tubules, the intracellular calcium stores. Such complex organization supports the fast stimulus-secretion coupling we described; localized calcium elevations have been recently observed in astrocytes in situ, suggesting that these functional microdomains might be present in the intact tissue. In the second part of the work, we investigated whether astrocytes possess some of the benchmarks of brain dopaminergic cells. It's been known for years that astrocytes are able to metabolize monoamines by the enzymes MAO and COMT, but to date no clear information that glial cells are able to uptake and store monoamines have been provided. Here, we identified a whole apparatus for the storage, degradation and release of monoamines, at the ultrastructural level. Electron microscopy immunohistochemistry allowed us to visualize VMAT2- and dopamine-positive intracellular compartments within astrocytic processes, i.e. dense -core granules and cisterns. These organelles might be responsible for dopamine release and storage, respectively; interestingly, this intracellular distribution is reminiscent of VMAT2 expression in dendrites if neurons, where dopamine release is tonic and plays a role in the regulation of its a basal levels, suggesting that astrocytic VMAT2 is involved in the homeostasis of dopamine in healthy brains of adult mammals.¦Durant cette dernière décennie, de nombreux résultats sur le relâchement des transmetteurs par les astrocytes pouvant modulé l'activité synaptique (gliotransmission) ont été fournis. Néanmoins, la gliotransmission reste un processus encore très débattu, notamment à cause de l'absence de preuves directes, morphologique et fonctionnelle démontrant ce phénomène. Nous présentons dans nos travaux de nombreux résultats confortant l'hypothèse de la gliotransmission, dont i) une étude approfondie sur les propriétés spatiales et temporelles de la sécrétion régulée du glutamate dans les astrocytes, et ii) une étude sur la participation des astrocytes dans la transmission de la dopamine, une neuromodulateur dont l'interaction avec les astrocytes est fortement probable, mais qui n'a encore jamais été prouvée. L'expression des petites vésicules (SLMVs - Synaptic Like Micro Vesicles) glutamatergiques exprimant les transporteurs vésiculaires du glutamate (VGLUTs) dans les astrocytes a déjà été prouvé tant in situ qu'in vitro. Afin de mettre en évidence les propriétés précises de la sécrétion de ces organelles, nous avons adapté à nos études des méthodes expérimentales conçues pour observer les processus de exocytose et endocytose dans les neurones. Les résolutions spatiale et temporelle obtenues, grâce a l'utilisation en parallèle de l'épi fluorescence et de la fluorescence a onde évanescente (TIRF), nous ont permis de montrer que la sécrétion régulée dans les astrocytes est un processus extrêmement rapide (de l'ordre de la milliseconde) et qu'elle est capable de soutenir et de suivre la transmission de signaux entre neurones. Nous avons également découvert que cette sécrétion a lieu dans des compartiments subcellulaires particuliers où nous observons la présence du reticulum endoplasmique (ER) ainsi que des augmentations rapides de calcium. Cette organisation spatiale complexe pourrait être la base morphologique du couplage rapide entre le stimulus et la sécrétion. Par ailleurs, plusieurs études récentes in vivo semblent confirmer l'existence de ces compartiments. Depuis des années nous savons que les astrocytes sont capables de métaboliser les monoamines par les enzymes MAO et COMT. Nous avons donc fourni de nouvelles preuves concernant la présence d'un appareil de stockage dans les astrocytes participant à la dégradation et la libération de monoamines au niveau ultrastructurelle. Grâce à la microscopie électronique, nous avons découvert la présence de compartiments intracellulaires exprimant VMAT2 dans les processus astrocytaires, sous forme de granules et des citernes. Ces organelles pourraient donc être responsables à la fois du relâchement et du stockage de la dopamine. De manière surprenante, cette distribution intracellulaire est similaire aux dendrites des neurones exprimant VMAT2, où la dopamine est libérée de façon tonique permettant d'agir sur la régulation de ses niveaux de base. Ces résultats, suggèrent une certaine participation des VMAT2 présents dans les astrocytes dans le processus d'homéostase de la dopamine dans le cerveau.¦A de nombreuses reprises, dans des émissions scientifiques ou dans des films, il est avancé que les hommes n'utilisent que 10% du potentiel de leur cerveau. Cette légende provient probablement du fait que les premiers chercheurs ayant décrit les cellules du cerveau entre le XIXème et le XXeme siècle, ont montré que les neurones, les cellules les plus connues et étudiées de cet organe, ne représentent seulement que 10% de la totalité des cellules composant du cerveau. Parmi les 90% restantes, les astrocytes sont sans doute les plus nombreuses. Jusqu'au début des années 90, les astrocytes ont été plutôt considérés peu plus que du tissu conjonctif, ayant comme rôles principaux de maintenir certaines propriétés physiques du cerveau et de fournir un support métabolique (énergie, environnement propre) aux neurones. Grace à la découverte que les astrocytes ont la capacité de relâcher des substances neuro-actives, notamment le glutamate, le rôle des astrocytes dans le fonctionnement cérébral a été récemment reconsidérée.¦Le rôle du glutamate provenant des astrocytes et son impact sur la fonctionnalité des neurones n'a pas encore été totalement élucidé, malgré les nombreuses publications démontrant l'importance de ce phénomène en relation avec différentes fonctions cérébrales. Afin de mieux comprendre comment les astrocytes sont impliqués dans la transmission cérébrale, nous avons étudié les propriétés spatio-temporelles de cette libération grâce à l'utilisation des plusieurs marqueurs fluorescents combinée avec différentes techniques d'imagerie cellulaires. Nous avons découvert que la libération du glutamate par les astrocytes (un processus maintenant appelé "gliotransmission") était très rapide et contrôlée par des augmentations locales de calcium. Nous avons relié ces phénomènes à des domaines fonctionnels subcellulaires morphologiquement adaptés pour ce type de transmission. Plus récemment, nous avons concentré nos études sur un autre transmetteur très important dans le fonctionnement du cerveau : la dopamine. Nos résultats morphologiques semblent indiquer que les astrocytes ont la capacité d'interagir avec ce transmetteur, mais d'une manière différente comparée au glutamate, notamment en terme de rapidité de transmission. Ces résultats suggèrent que le astrocytes ont la capacité de modifier leurs caractéristiques et de s'adapter à leur environnement par rapport aux types de transmetteur avec lequel ils doivent interagir.