Follicular epithelium, theca and egg envelope formation in Serrasalmus spilopleura (Teleostei, Characiformes, Characidae)

The follicular epithelium and theca of oocytes in Serrasalmus spilopleura differentiates during the initial primary growth phase. The follicular cells are squamous and the thecal cells are disposed in two layers. During the secondary growth phase, follicular cells become cuboidal, acquire characteristics typical of protein- or glycoprotein-producing cells, and show dilated intercellular spaces. Formation of the egg envelope in S. spilopleura begins in the previtellogenic oocytes as a layer of amorphous electron-dense material is laid down on the oolemma. During vitellogenesis, another layer of electron-dense material appears beneath the first layer. Also during this phase, a layer of amorphous, less electron-dense material is formed adjacent to the follicular epithelium. The secondary egg envelope appears at the postvitellogenic phase and is composed of a filamentous and undulant material. The morphology of the egg envelopes in S. spilopleura reflects not only its oviparous nature but also the fact that its eggs are adhesive.

Morphological investigations of the surface epithelium of ductuli efferentes of black isogenic mice (Mus musculus)

Morphological investigations of the epithelial cell types that line the ductuli efferentes (DE) of black isogenic mice confirm absorption of the luminal fluid phase by endocytosis as the main function of ductuli efferentes (DE) in this species. :Furthermore, all the histochemical and ultrastructural observations on the DE epithelial histoarchitecture indicate other cellular functions such as exocytosis and probably secretion: including an aprocrine secretory process.

Morphometry of the midgut epithelium of Diatraea saccharalis Fabricius, 1794 (Lepidoptera) parasitized by Cotesia flavipes Cameron, 1891 (Hymenoptera)

The morphometric study of the midgut in Diatraea saccharalis (Lepidoptera) larvae parasitized by the Cotesia flavipes (Hymenoptera) showed that there was significant increase in the columnar, goblet and regenerative cells and their nuclei; the midgut lumen diameter and the epithelial height were also increased in the parasitized larvae. The multivariate analysis showed that parasitism affected the columnar cell only in the posterior region, and the goblet cells along the midgut length (anterior and posterior regions). (c) 2006 Elsevier B.V. All rights reserved.

Morphometric study of the midgut epithelium in larvae of Diatraea saccharalis Fabricius (Lepidoptera: Pyralidae)

A larva de Diatraea saccharalis Fabricius (broca-da-cana) tem grande interesse econômico, pois afeta o cultivo e aproveitamento industrial da cana-de-açúcar. Entretanto, poucos são os estudos sobre a morfologia interna desse inseto. O objetivo deste trabalho foi estudar, morfometricamente, o seu epitélio intestinal, ao longo de seu comprimento, visando caracterizar regiões estruturalmente diferentes. O intestino médio de larvas no último instar foi subdividido em três regiões: proximal, mediana e distal e os fragmentos foram processados para observação em microscopia de luz. Os cortes histológicos foram analisados em sistema computadorizado de análise de imagens para medir comprimento, largura e área do epitélio, das diferentes células epiteliais, dos seus respectivos núcleos e do lúmen intestinal. Os dados obtidos foram submetidos ao teste estatístico de Kruskal-Wallis e à análise multivariada. Nossos resultados mostraram que o intestino médio apresentou-se constituído, morfometricamente, por duas diferentes regiões, proximal e distal; a região mediana apresentou valores coincidentes tanto com a região proximal quanto com a distal, sugerindo ser região intermediária. As células epiteliais (colunares, caliciformes e regenerativas), quando avaliadas pela análise estatística multivariada, não apresentaram diferença morfométrica nas diferentes regiões do intestino médio. Entretanto, a análise de variância, realizada para variáveis isoladas, mostrou que as células regenerativas apresentaram maior variabilidade morfométrica.

Germ line Cysts and the Formation of the Germinal Epithelium During the Female Gonadal Morphogenesis in Cyprinus Carpio (Teleostei: Ostariophysi: Cypriniformes)

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

Giardia agilis: Ultrastructure of the Trophozoites in the Frog Intestine

Intestine samples of Bufo sp. tadpoles with parasitism confirmed for Giardia agilis were studied by transmission electron microscopy. The G. agilis trophozoites were long and thin. The plasma membrane was sometimes undulated and the cytoplasm, adjacent to the dorsal and ventral regions, showed numerous vacuoles. The two nuclei presented prominent nucleoli. The cytoplasm was electron-dense with free ribosomes, glycogen and rough endoplasmic reticulum-like structures. Polyhedral inclusions were observed in the cytoplasm and outside the protozoan; some of these inclusions exhibited membrane disruption. The flagella ultrastructure is typical, with the caudal pair accompanied by the funis. Next to the anterior pair, osmiophilic material was noticed. The ventro-lateral flange was short and thick, supported by the marginal plates that penetrated into its distal extremity; only its distal portion had adjacent osmiophilic filament. The G. agilis trophozoites showed the general subcellular feature of the genus. However, the ventro-lateral flange ultrastructure was an intermediate type between G. muris and G. duodenalis.

Identification and characterization of excretory/secretory products of Giardia duodenalis trophozoites

One of the major questions concerning Giardia is the understanding of pathophysiological processes associated with small intestine abnormalities. There are evidences that Giardia trophozoites contain and/or release proteolytic enzymes that may be implicated in the host intestinal epithelium. The present investigation was undertaken to examine the protease activity in excretory/secretory (E/S) products of Giardia duodenalis trophozoites of an axenic Brazilian strain (BTU-11) and the reference strain Portland 1 (P1). E/S products from trophozoites of each strain in conditioned medium were tested with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for the protein profiles, and the protease activity was analyzed using substrate-impregnated SDS-PAGE (gelatin and collagen) and hemoglobin assay. The proteases characterization was based on inhibition assays including synthetic inhibitors. Electrophoresis analysis of E/S products revealed a banding pattern composed by few bands (4 to 6 bands) in the migration region of 123 to 28 kDa. Proteolytic products were detected in the conditioned medium by trophozoites of both assayed strains. In the gels containing copolymerized gelatin and collagen, E/S products promoted substrate degradation and the most evident proteolysis zones were distributed in the migration regions of 77 to 18 kDa and 145 to 18 kDa, respectively, in the patterns of gelatinolytic and collagenolytic activities. Degradation of hemoglobin was also observed, and the pattern of hydrolysis was similar in both E/S products assayed. Inhibitor assays showed that the main proteolytic activity in both E/S products is due to cysteine proteases, although the presence of serine proteases was also indicated. Degradation of substrates including collagen and hemoglobin could lead us to speculate different functions of Giardia excreted/secreted proteases in vivo, but to confirm this possibility and to elucidate its implication on host-parasite interactions, further experiments applying protocols for the purification of proteases are necessary. Even so, our observations are relevant and hold the perspective for the understanding about protease activity in Giardia trophozoites of axenic strain isolated in an endemic area.

Ultrastructural study of the follicular epithelium in oocytes of Pachycondyla (Neoponera) villosa ants (Hymenoptera : Ponerinae)

Insect oocytes are surrounded by the follicular epithelium which is simple and cuboidal, wih the mainly functions of: synthesis of vitellin membrane and chorion and synthesis and transport of hemolymph products (proteins). In Pachycondyla (Neoponera) villosa ants workers aged less than 10 days do not present the formation of ovarian follicles (oocytes, nurse cells and follicular cells) indicating that vitellogenesis starts at approximately 10 days of age. Studies of participation of the follicular epithelium in Pachycondyla (Neoponera) villosa showed that in stage I oocytes the epithelium does not present the opening of intercellular spaces. In stage II these spaces begin to be observed together with separation of the follicular epithelium from the oocyte surface. In stage III two types of material were observed in the intercellular spaces: electrodense material in the basal region and compacted material in apical one as well as follicular epithelium/oocytes interface suggesting that the extraovarian material that reach oocytes undergoes some type of modification during passage through the intercellular spaces. The follicular epithelium spaces in queen are bigger than in workers oocytes.

Ultrastructure of the midgut epithelium of Meliponinae larvae with different developmental stages and diets

The comparative study of the ultrastructure of the midgut epithelium of stingless bee larvae that eat plant protein (pollen) and animal protein (carrion) throughout the larval phase, shows variations in the digestive cells that are only relative to larval aging and not to the type of larval diet. The cells of older larvae present a cytoplasm with empty spaces that result from emptying of lipid and glycogen stocks, and the presence of autophagic vacuoles. These results are discussed in relation to the hypothesis that variations in the digestive tract of insects may be associated with different diets or phylogeny. We conclude that different diets do not determine cell morphology adaptations in the studied species. As the variations in the ultrastructure of the midgut epithelium are the same in all studied species, including the necrophagous species Trigona hypogea, throughout the larval stage, this sequence of changes seems to be due to different physiological state during larval development.

The midgut of Cephalotes ants (Formicidae: Myrmicinae): Ultrastructure of the epithelium and symbiotic bacteria

The ultrastructural analysis of the midgut of Cephalotes atratus. C. clypeatus, and C. pusillus reveled that the midgut epithelium lays on a basal lamina and is composed basically of three cell types: digestive cells, regenerative cells, and goblet cells. In these ants, the rough endoplasmic reticulum, in addition to producing digestive enzymes, is involved in the formation of concretions and ion storage in specialized vacuoles present in the midgut. These concretions are spherocrystals and may contribute to stabilize the pH and to maintain symbiotic bacteria found between microvilli. The ultrastructure analysis of these bacteria revealed the presence of a double envelope typical of gram-negative bacteria. For the three species examined, the ultrastructure similarities are conspicuous, suggesting that this may be the pattern for the genus Cephalotes. Details of the relationship between bacteria and microvilli were examined. (C) 2010 Elsevier Ltd. All rights reserved.

Histology and Histochemistry of the Stomach Epithelium of Pachycondyla (= Neoponera) villosa (Formicidae: Ponerinae)

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

Female reproductive system of the sugarcane spittlebug Mahanarva fimbriolata (Auchenorrhyncha): Vitellogenesis dynamics and protein quantification

The present study aimed describing the ovaries of the sugarcane spittlebug Mahanarva fimbriolata which are meroistic telotrophic with nurse cells and oocytes located in the tropharium. SEM revealed paired ovaries located dorsolaterally around the intestine, and oocytes exhibiting shapes ranging from round (less developed) to elliptic (more developed), suggesting a simultaneous, although, asynchronous development. Based on histological data we classified the oocytes in stages from I to V. Stage I oocytes exhibit follicular epithelium with cubic and/or prismatic cells, fine cytoplasmic granules. Stage II oocytes present intercellular spaces in the follicular epithelium due to the incorporation of yolk elements from the hemolymph. Small granules are present in the periphery of oocytes while larger granules are observed in the center. Stage III oocytes are larger and intercellular spaces in the follicular epithelium are evident, as well as the interface between follicular epithelium and oocyte. Yolk granules of different sizes are present in the cytoplasm. During this stage, chorion deposition initiates. Stage IV oocytes exhibit squamous follicular cells and larger intercellular spaces when compared to those observed in the previous stage. The oocyte cytoplasm present granular and viscous yolk, the latter is the result of the breakdown of granules. Stage V oocytes exhibit a follicular epithelium almost completely degenerated, smaller quantities of granular yolk and large amounts of viscous yolk. Based on our findings we established the sequence of yolk deposition in M. fimbriolata oocyte as follows: proteins and lipids, which are first produced by endogenous processes in stages I and II oocytes. Exogenous incorporation begins in stage III. In stages I and II oocytes, lipids are also produced by follicular epithelial cells. The third element to be deposited is polysaccharides, mainly found as complexes. Therefore, the yolk present in the oocytes of this species consists of glycolipoproteins. Molecular weights of proteins present in M. fimbriolata oocytes ranged from 10 to 92 KDa, differently from vitellogenin, the most common protein present in insect oocytes, weighing approximately 180 KDa. (c) 2006 Elsevier Ltd. All rights reserved.

Structure, histochemistry, and ultrastructure of the epithelium and stroma in the gerbil (Meriones unguiculatus) female prostate

The female prostate has aroused scientific interest because it is subjected to the same diseases compromising the male prostate during aging. The objective of this work was to characterize structurally, cytochemically, and ultrastructurally the tissue compartments of the normal adult female prostate of Meriones unguiculatus gerbils. The morphological analyses showed that the gerbil's female prostate is constituted of a cluster of glands and ducts inserted in a musculofibrous stroma. The alveolar epithelium is differentiated and consisted of basal proliferating cells, intermediary cells, and secretory cells. The secretory cells are the most numerous cell type and continuously secrete glycoproteins. The basal cells are the source of the secretory cells and they are then responsible for the alveolus renovation. The prostatic stroma is abundant and rich in elastic and collagen fibers, which are closely associated with smooth muscle cells and fibroblasts. The results showed that the gerbil's female prostate shows morphological and ultrastructural homology to the human female prostate (Skene's gland), and despite being a small organ, it is a mature and physiologically active gland. (C) 2003 Elsevier Ltd. All rights reserved.

Regeneration of the Corneal Epithelium after Debridement of its Central Region: An Autoradiographic Study on Rabbits

Purpose: To investigate the proliferative behavior of the corneal and limbal epithelia after debridement on the central region of the rabbit cornea. Methods: After scraping a circular epithelial area, 5 mm in diameter, in the center of the cornea, (3)H-thymidine ((3)H-TdR) was injected intravitreally, and the rabbits killed from 1 to 49 days afterward. The cornea, together with the adjacent conjunctiva, was processed for autoradiography. Results: The regenerating epithelium at the center of the cornea exhibited high frequencies of labeled nuclei when compared to controls. The mitotic indexes for the limbus were comparable in experimental and control eyes. The unique basal stratum of the limbal epithelium exhibited quick proliferation and vertical migration in all eyes. Cells that remained labeled for four weeks or more were observed throughout the corneal epithelium, including its basal stratum, and this did not depend on epithelial damage. Conclusion: Corneal epithelium wounds are healed by sliding and proliferation of cells surrounding the epithelial gap without any evidence for the participation of the limbal epithelium. Daughter cells labeled with (3)H-TdR were visualized in all layers of the corneal epithelium up to 7 weeks after the DNA precursor injection. However, at this long interval, the only labeled cells in the limbus were in the suprabasal layers.