Ultrastructure of ovarian follicular epithelium of the Amazonian fish Pseudotylosurus microps (Teleostei: Belonidae): morphological and histochemical characterization of the intercellular deposits

Neste trabalho é mostrada a grande quantidade de material eletrondenso intercelular no epitélio folicular de P. microps. Aparentemente, o material é captado da circulação e enviado para o folículo por meio dos espaços intercelulares, acumulando-se nos espaços intercelulares médio-apicais do epitélio e no espaço perioocítico. A acumulação é iniciada no oócito primário e prossegue até a vitelogênese. A natureza química desse material é discutida.

Urocortin in the central nervous system of a primate (Cebus apella): Sequencing, immunohistochemical, and hybridization histochemical characterization

The urocortin (UCN)-like immunoreactivity and UCN mRNA distribution in various regions of the nonprimate mammalian brain have been reported. However, the Edinger-Westphal nucleus (EW) appears to be the only brain site where UCN expression is conserved across species. Although UCN peptides are present throughout vertebrate phylogeny, the functional roles of both UCN and EW remain poorly understood. Therefore, a study focused on UCN system organization in the primate brain is warranted. By using immunohistochemistry (single and double labeling) and in situ hybridization, we have characterized the organization of UCN-expressing cells and fibers in the central nervous system and pituitary of the capuchin monkey (Cebus apella). In addition, the sequence of the prepro-UCN was determined to establish the level of structural conservation relative to the human sequence. To understand the relationship of acetylcholine cells in the EW, a colocalization study comparing choline acetyltransferase (ChAT) and UCN was also performed. The cloned monkey prepro-UCN is 95% identical to the human preprohormone across the matched sequences. By using an antiserum raised against rat UCN and a probe generated from human cDNA, we found that the EW is the dominant site for UCN expression, although UCN mRNA is also expressed in spinal cord lamina IX. Labeled axons and terminals were distributed diffusely throughout many brain regions and along the length of the spinal cord. of particular interest were UCN-immunoreactive inputs to the medial preoptic area, the paraventricular nucleus of the hypothalamus, the oral part of the spinal trigeminal nucleus, the flocculus of the cerebellum, and the spinal cord laminae VII and X. We found no UCN hybridization signal in the pituitary. In addition, we observed no colocalization between ChAT and UCN in EW neurons. Our results support the hypothesis that the UCN system might participate in the control of autonomic, endocrine, and sensorimotor functions in primates.

Histochemical study of the extensor digitorum longus and soleus muscles in alcoholic rats

The extensor digitorum longus (EDL) and soleus (SOL) muscle fibres from albino rats submitted to experimental chronic alcoholism were evaluated in accordance with their metabolic and morphometric profiles. Twenty-seven male animals aged 4 months and weighing approximately 400 g were used. The animals were divided into three groups: control, isocaloric and alcoholic and sacrifices were carried out after 5, 10 and 15 months. The muscles were dissected, removed, cross-sectioned in a cryostat and submitted to the NADH (nicotinamide adenine dinucleotide) reaction. The SO (slow-twitch-oxidative), FG (fast-twitch-glycolytic) and FOG (fast-twitch-oxidative-glycolytic) muscle fibre types exhibited a polygonal, triangular or rounded shape and did not present noteworthy modifications in either muscles during the study. The cross-sectional areas of the fibres from the studied muscles did not present significant differences during the observations. Fibre area behaved similarly in the alcoholic animals up to the 10th month, i.e. it was decreased, as also observed in the other groups. At 15 months, however, all fibres were increased, with a predominance of FG fibres in the SOL muscle. Changes in fibre population were observed mainly in the SOL muscle of alcoholic animals: SO fibres were initially increased in number but decreased after the 10th month, and the opposite was observed for the population of FG fibres. FOG fibres increased linearly in number throughout the experiment. The statistical analysis showed nevertheless that the fibre population and cross-sectional area changes were not significant. In the alcoholic animals quantitative variations of muscle fibres were more evident in the SOL muscle, suggesting that the SOL muscle is more sensitive to the toxic action of ethanol. The results concerning the increased fibre diameter in alcoholic animals would be associated with muscle oedema induced directly or indirectly by the ethanol.

Ultrastructural and histochemical examination of alveolar bone at the pressure areas of rat molars submitted to continuous orthodontic force

It is usually believed that repair in alveolar bone during orthodontic movement occurs after decreasing of force. However, we have recently observed signs of repair in previously resorbed cementum from human teeth exposed to continuous forces. In order to test the hypothesis that bone resorption and deposition occur concomitantly at the pressure areas, a continuous 15 cN force was applied in a buccal direction to upper first molars from eight 2.5-month-old male Wistar rats for 3 d (n=4) and 7 d (n=4). As a control, two additional rats did not have their molars moved. Maxillae were fixed in 2% glutaraldehyde + 2.5% formaldehyde, under microwave irradiation, decalcified in ethylenediaminetetraacetic acid, and processed for transmission electron microscopy. Specimens from one rat from each group were processed for tartrate-resistant acid phosphatase (TRAP) histochemistry. At both the times studied, the alveolar bone surface at the pressure areas showed numerous TRAP-positive osteoclasts, which were apposed to resorption lacunae. In addition, osteoblasts with numerous synthesis organelles were present in the neighboring areas overlying an organic matrix. Thus, this study provides evidence that the application of continuous forces produces concomitant bone resorption and formation at the pressure areas in rat molars.

Histochemical and ultrastructural cytochemistry of glycogen on ovarioles of Neoponera villosa ants (Hymenoptera: Ponerinae)

In Neoponera villosa ants, we found ovaries of the polytrophic meroistic type which is characterized by the presence of nurse cells forming together with the oocyte, the so-called follicles. The nurse cells have the primary function of supplying the oocyte with RNA, but they contribute to the supply of other elements such as glycogen. With the objetive of detecting the presence of this substance in the ovarioles of workers and queens of N.villosa ante the ovaries were removed and processed according to electron microscopy technic for glycogen detection. Glycogen is a common element in insect oocytes and is abundantly distributed in the cytoplasm of N.villosa workers and queens. However, in ovarian follicles it can only be detected at stages ET and lit of development. Glycogen synthesis probably occurs predominantly in nurse cells which transfer it into the oocyte through the nourish pore. This process requires high energy expenditure that justify the large numbers of mitochondria associated with glycogen in the nurse cell cytoplasm. The amount of glycogen in the nurse cells of queens is slightly greater than workers.

Morphometric and histochemical study of the human vocal muscle

The present study was conducted on vocal muscles removed at autopsy Rom adult individuals (10 men and 8 women, aes ranging from 48 to 78 years) with no laryngeal disease. Histologic analysis was performed with hematoxylin and eosin staining, and histochemical analysis was performed by nicotinamide-adenine-dinucleotide tetrazolium reductase, succinate dehydrogenase, and acid and alkaline myofibrillar adenosine triphosphatase reactions. The histochemical reactions showed that the muscle consists of slow-twitch oxidative (SO), fast-twitch glycolytic (FG), and fast-twitch glycolytic oxidative (FOG) fibers distributed in mosaic form. The frequencies of SO, FOG, and FG fibers were 40.50%, 54.75%, and 4.75%, respectively. The higher frequency of SO and FOG oxidative fibers characterizes the muscle as having aerobic metabolism, resistance to fatigue, and fast contraction. The mean minimum diameters were 31.37 mu m for SO fibers and 36.46 mu m for FOG and FG fibers.

HISTOCHEMICAL RESPONSES of RATS EXERCISED IN TWO WEEKLY FREQUENCIES and INGESTING STANDARD OR HYPERCALORIC DIET

This study investigated if overfed rats present morphological and histochemical muscle adaptation similar to normally fed, both submitted to two different weekly frequencies of training. Thirty male Wistar rats were fed either with standard chow (SCO) or with hypercaloric diet (HCO). They were subdivided into six subgroups: sedentary (SCO and HCO), trained twice/week (SC2 and HC2) and trained five times/week (SC5 and HC5). The trained groups swam 60 min/day, during 10 weeks. Twenty four hours after the last training, samples of Gastrocnemius were excised and stained with HE, NADH-TR and m-ATPase, and the capillary density was calculated. Total heart mass (HM) and the mass of atrium (AM), left (LV) and right (RV) ventricles were excised and weighted. The comparisons were made by ANOVA and by Covariance analysis, adjusting the variables by body weight. The results showed that the HCO achieved higher BM, however, absolute HM did not differ post training. Irrespective of the diet, rats that were trained twice a week presented significantly greater increase in the AM. In general, the SC5 and HC5 groups showed higher HM, LV, RV, proportion of oxidative fibres and capillary density, compared to the sedentary and twice week trained groups. A higher proportion of injuries (splitting) was noted in the HC2 and HC5 compared to SC2 and SC5. These results indicate that the frequency of training influenced the skeletal and heart adaptation and larger changes were observed in the 5x/week group, which ingested the standard diet. The 5x/week training groups also presented large amount of muscle fibres damage.

Histochemical and SEM evaluation of the neuromuscular junctions from alcoholic rats

In the present study morphological changes occurring in the neuromuscular junctions (NMJ) of the extensor digitorum longus (EDL) and soleus muscles from albino rats (Rattus norvegicus) submitted to experimental chronic alcoholism were evaluated. Seventy two male animals aged 4 months and weighing on average 400 g were divided into three groups: control, alcoholic and isocaloric. Six rats from each group were anesthetized and sacrificed after 5, 10, 15 and 18 months. The NMJ did not show detectable morphological changes in either muscle after treatment when examined by light microscopy. With respect to the dimensions, statistical analysis demonstrated a tendency to a statistically significant treatment x time interaction for the length of soleus muscle NMJ. The ultrastructural study, however, revealed that the NMJ of the soleus muscle of animals submitted to 18 months of experimental alcoholism presented important morphological alterations. Characteristically, the NMJ of these muscles is located on an elevation on the surface of the muscle fiber, presenting a regular round, oval or elliptical shape and continuous and not very deep synaptic grooves. Approximately 30% of the NMJ of alcoholic rats are irregular in shape, with the sarcolemmal elevations typical of the synapse region being flattened on at least one side, with discontinuous synaptic grooves, and deep and punctiform contacts of the synaptic buds. These data suggest that, although skeletal muscle has a greater natural resistance against the direct or indirect effects of alcohol, some submicroscopic morphological alterations are detectable in the NMJ, especially in muscles with oxidative metabolism (soleus) following long periods of ingestion of alcohol. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Morphological and Histochemical Characterization of Gill Filaments of the Brazilian Endemic Bivalve Diplodon expansus (Kuster, 1856) (Mollusca, Bivalvia, Hyriidae)

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

Glycogen distribution in ameloblasts and odontoblasts of newborn rats of sialoadenectomized dams. Histochemical study

In order to get information about the distribution of glycogen in ameloblasts and odontoblasts, studies were made of newborn rats of sialoadenectomized dams and newborn rats of control dams. Rodent offspring were decapitated on the 1st, 3rd, 5th, 7th, and 9th days after birth. Their heads were fixed in 10% neutral formalin solution, decalcified in sodium citrate-formic acid and embedded in paraffin, and frontal sections were prepared. Sections 6 micrometers thick were stained by specific histochemical reactions to detect glycogen. Based on the results obtained, it was concluded that the amount of glycogen was lower in the cytoplasms of ameloblasts and odontoblasts of experimental animals when compared to controls.