Programmed cell death in the larval salivary glands of Apis mellifera (Hymenoptera, Apidae)

The morphological and histochemical features of degeneration in honeybee (Apis mellifera) salivary glands were investigated in 5th instar larvae and in the pre-pupal period. The distribution and activity patterns of acid phosphatase enzyme were also analysed. As a routine, the larval salivary glands were fixed and processed for light microscopy and transmission electron microscopy. Tissue sections were subsequently stained with haematoxylin-eosin, bromophenol blue, silver, or a variant of the critical electrolyte concentration (CEC) method. Ultrathin sections were contrasted with uranyl acetate and lead citrate. Glands were processed for the histochemical and cytochemical localization of acid phosphatase, as well as biochemical assay to detect its activity pattern. Acid phosphatase activity was histochemically detected in all the salivary glands analysed. The cytochemical results showed acid phosphatase in vesicles, Golgi apparatus and lysosomes during the secretory phase and, additionally, in autophagic structures and luminal secretion during the degenerative phase. These findings were in agreement with the biochemical assay. At the end of the 5th instar, the glandular cells had a vacuolated cytoplasm and pyknotic nuclei, and epithelial cells were shed into the glandular lumen. The transition phase from the 5th instar to the pre-pupal period was characterized by intense vacuolation of the basal cytoplasm and release of parts of the cytoplasm into the lumen by apical blebbing; these blebs contained cytoplasmic RNA, rough endoplasmic reticule and, occasionally, nuclear material. In the pre-pupal phase, the glandular epithelium showed progressive degeneration so that at the end of this phase only nuclei and remnants of the cytoplasm were observed. The nuclei were pyknotic, with peripheral chromatin and blebs. The gland remained in the haemolymph and was recycled during metamorphosis. The programmed cell death in this gland represented a morphological form intermediate between apoptosis and autophagy.

Nuclear alterations associated to programmed cell death in larval salivary glands of Apis mellifera (Hymenoptera : Apidae)

The silk glands of bees are a good model for the study of cell death in insects. With the objective to detect the nuclear features during glandular regression stage, larvae at the last instar and pre-pupae were collected and their silk glands were dissected and processed for ultrastructural analysis and histologically for cytochemical and imunocytochemical analysis. The results showed that the cellular nuclei exhibited characteristics of death by atypical apoptosis as well as autophagic cell death. Among the apoptosis characteristic were: nuclear strangulation with bleb formation in some nuclei, DNA fragmentation in most of the nuclei and nucleolar fragmentation. Centripetal chromatin compaction was observed in many nuclei, forming a perichromatin halo differing from typical apoptotic nuclei. With regards to the characteristics of autophagic-programmed cell death, most relevant was the delay in the collapse of many nuclei. (C) 2006 Elsevier Ltd. All rights reserved.

Cytochemistry of fat body trophocytes and ovaries of workers and queens of Melipona quadrifasciata anthidioides (Hymenoptera: Apidae: Meliponini) during vitellogenesis

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

Histochemistry and Protein Profile of The Venom Glands of Workers of Neoponera villosa Ants (Hymenoptera: Ponerinae)

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

Death by apoptosis in salivary glands of females of the tick Rhipicephalus sanguineus (Latreille, 1806) (Acari : Ixodidae)

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

Fat body cells of Amblyomma cajennense partially engorged females (Acari: Ixodidae) and their role on vitellogenesis process

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ultrastructural features of the hypopharyngeal glands in the social wasp Polistes versicolor (Hymenoptera : Vespidae)

The wasps of the genus Polistes have been considered the key to understanding the evolution of social behavior in Hymenoptera. Several studies have shown that the development of organized insect societies was accompanied by the evolution of structures like exocrine glands, which became specialized to perform specific functions. This article investigates the ultrastructural and cytochemical features of the hypopharyngeal glands of Polistes versicolor. These glands have been studied in depth in social bees, where they occur only in nurses and produce the royal jelly. Our results revealed that these glands basically did not vary among individuals or between sexes. They are constituted by spherical cells, each with a large nucleus and well-developed rough endoplasmic reticulum. Secretion vesicles are abundant, but lipid droplets were not observed, indicating that these glands may not have a role in pheromone synthesis. Acid phosphatase was detected in lysosomes, and also free in the cytosol, but did not seem to be related with cell death. Thus, our results suggest that the hypopharyngeal glands of P. versicolor may not have a specialized social role, but could produce digestive enzymes.

Nucleolar Cycle and Its Correlation with Chromatoid Bodies in the Tilapia rendalli (Teleostei, Cichlidae) Spermatogenesis

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

Nucleolar cycle and chromatoid body formation Is there a relationship between these two processes during spermatogenesis of Dendropsophus minutus (Amphibia, Anura)?

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Mg2+-dependent ATPase activity in Malpighian tubules of Triatoma infestans Klug

Mg2+-dependent ATPases were investigated in Malpighian tubules of the blood-sucking insect, Triatoma infestans, with cytochemical procedures for light and electron microscopy. The aim was to establish patterns of enzyme occurrence in the blood-sucking insect under control rearing conditions for further comparisons with animals subjected to the action of stress factors. Enzyme activity was found in laminated "concretions" present in distal cells, in edges of urate crystals at the lumen of the proximal region of tubules, in the basement membrane of proximal cells, and variously distributed in plasmalemma invaginations of both distal and proximal cells. Presence of ATPases in the "concretions" and urate crystals is presumed to be due to engulfment of other ATPase-containing components during formation of these structures. Cytochemical reactivity in the basement membrane and plasmalemma invaginations is assumed to be involved with active transport of waste molecules from and to hemolymph and differs as a function of the Malpighian tubule region. This paper provides a basic understanding of the enzyme occurrence in the blood sucking insects, and can be used as a pattern for comparative means of the staining patterns among Triatominae species. (C) 2011 Elsevier Ltd. All rights reserved.

Study of the nucleolar cycle and ribosomal RNA distribution during meiosis in triatomines (Triatominae, Heteroptera)

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

Relationship between the nucleolar cycle and chromatoid body formation in the spermatogenesis of Phrynops geoffroanus (Reptilia Testudines)

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

Dynamics and cytochemistry of oogenesis in Astyanax fasciatus (Cuvier) (Teleostei, Characiformes, Characidae) from Rio Sapucaí, Minas Gerais State, Brazil

Oogenesis involves a set of transformations which are undergone by female germ cells These cells change into oogonias and then into mature oocytes. Sexually mature female fish were collected monthly, during one year, from the Sapucaí River, a tributary of the Rio Grande, which is part of the Furnas Reservoir in the state of Minas Gerais. During the several stages of maturation, we observed small round oogonias with a large nucleus, a single nucleolus, and weakly stained cytoplasm with eosinophilic granules. The primary oocytes showed a large basophilic nucleus, with a developed peripheral nucleolus and a reduced cytoplasm. The previtellogenic oocytes presented voluminous cytoplasm and nucleus with several small peripheral nucleoli. The oocytes underwent vitellogenesis with the development of the zona radiata and the follicle cells. Their cytochemical reactions indicated that the two layers of the zona radiata of A. fasciatus contained proteins and polysaccharides. The initially squamous follicle cells, became cuboidal. In mature oocytes, the nucleus moved toward the periphery, next to the micropyle, and the yolk granules formed by proteins, fulfilled the cytoplasm. The clear unstained vesicles are likely to be the cortical alveoli in the perivitelline region.

Dynamics and cytochemistry of oogenesis in Leporinus striatus Kner (Teleostei, Characiformes, Anostomidae) from the Rio Sapucaí, Minas Gerais State, Brazil

Oogenesis involves a sequense of transformations which are undergone by female germ cells. These cells change into oogonias and then into mature oocytes. Sexually mature females were collected monthly, during one year, from the Rio Sapucaí, tributary of the Rio Grande, which is part of the Furnas Reservoir system in the state of Minas Gerais. The observed material showed that oogonias were small spherical cells, had a big spherical nucleus, with a single nucleolus, and weakly stained cytoplasm with eosinophilic granules (FG stained), which indicate their protein content. The primary oocytes showed a big basophilic nucleus, with a large peripheral nucleolus, and several smaller nucleoli. They show a reduced cytoplasmic content. The previtellogenic oocytes presented voluminous cytoplasm and nucleus with several small peripheral nucleoli. The oocytes underwent vitellogenesis with the development of the zona radiata and the follicle cells. The zona radiata had two layers, the outer and the inner, which showed its protein content when stained with CM and FG techniques. TB pH 2.5 and pH 4.0 staining showed that oocytes undergoing vitellogenesis presented weakly stained cytoplasm and peripheral cytoplasmic vesicles. The follicle cells that were squamous became cuboidal. In mature oocytes, the yolk granules that filled the cytoplasm became green and blue when stained with FG and CM techniques, indicating their protein content. The perivitclline region showed rosy stained vesicles (TB pH 2.5 and pH 4.0) spread among the weakly stained peripheral vesicles, which seemed to be the cortical alveoli. The zona radiata cells, CM and FG stained, still showed two layers like the oocytes from the previous stage, but thicker.