Morphometric changes associated with sex and development in the Malpighian tubules of Aedes aegypti

The Malpighian tubules of Aedes aegypti showed significant differences in their diameters between male and female larvae, male and female pupae, male larvae and male adults and male pupae and male adults. In every case, female values were greater than in males. Measurements of mean nuclear areas of the principal and stellate cells from Malpighian tubules, taken in males and females during development, showed that this parameter in both types of cell was significantly greater in females than in male larvae, pupae and adult stages. In males, significant differences between developmental stages were observed only in comparison with the nuclear area of larvae and adults in the principal cells, but in females, every comparison between stages showed significant differences except between pupae and adults in stellate cells. The frequency distribution of nuclear area values, in development, for male stellate and principal cells, were mostly concentrated in the first seven classes among the 30 classes considered in every stage, while for females, the frequency dropped drastically in the same classes from larvae to pupae and adults, moving to classes of higher values. Considering the importance of Malpighian tubules in insect physiology, the meaning of the differences detected are discussed on the basis of different metabolic levels, between sexes and developmental stages.     

Ultrastructure of intestinal and gall-bladder epithelium in the teleost Gasterosteus aculeatus L., as related to their osmoregulatory function

Summary Intestinal and gall-bladder epithelial cells in sticklebacks have been examined in ultrathin sections and freeze-etch replicas. Enterocytes throughout the intestine appear to have a well-developed basal labyrinth similar to that of renal tubular cells, consisting of baso-lateral infoldings closely associated with numerous mitochondria. The lumen inside these intracellular membranes is continuous with the intercellular space via pores. Such a membrane system is also present in the epithelial cells lining the gall bladder, distinguishing them from gall-bladder cells of higher vertebrates. Morphometric analysis indicates that the basal labyrinth of enterocytes in the posterior part of the intestine increases markedly in both sexually mature males and androgen-treated females. This does not occur in the anterior part or gall bladder. In sticklebacks, androgens cause reduced urine excretion and enhanced fluid release via the anus. We conclude that the cells lining the intestine and gall bladder possess an extensive basal labyrinth that may function as a backward channel system, enabling fluid to be produced in the intestine of fish. The androgen-induced increase in the extent of the basal labyrinth in the posterior part of the intestine may be related to the enhanced rate of intestinal fluid excretion observed in sexually mature male sticklebacks.     

Digestive cells in the midgut of Triatoma vitticeps (Stal, 1859) in different starvation periods

Triatoma vitticeps (Stal, 1859) is a hematophagous Hemiptera that, although being considered wild, can be found in households, being a potential Chagas’ disease vector. This work describes the histology and ultrastructure of the midgut of T. vitticeps under different starvation periods. Fifteen adults of both sexes starved for 3, 7, 20 and 25 days were studied. In general, digestive cells had apical microvilli, basal plasma membrane infoldings and central nucleus. The perimicrovillar membrane was found in all insects examined. Digestive cells of anterior midgut had lipid droplets, glycogen granules, developed basal labyrinth associated with mitochondria suggesting their role in nutrient storage and in fluid and ion transport. The cells of median and posterior regions of the midgut were rich in rough endoplasmic reticulum, lysosomes, vesicles and granules with different electron-densities. Moreover, cells of the posterior portion of the midgut had hemozoyn granules and mitochondria in the apical cytoplasm close to microvilli, suggesting their role in blood digestion and active nutrient absorption. The midgut of T. vitticeps showed differences in digestive cells associated with the time after feeding, and the increase of vesicles amount in long starvation periods, which suggests enzyme storage, which is readily used after a blood meal.     

Ultrastructure of midgut endocrine cells in the adult mosquito, Aedes aegypti

The ultrastructure of endocrine cells in the midgut of the adult mosquito, Aedes aegypti, resembled that of endocrine cells in the vertebrate gastro-intestinal tract. Midgut endocrine cells, positioned basally in the epithelium as single cells, were cone-shaped and smaller than the columnar digestive cells. The most distinctive characteristic of endocrine cells was numerous round secretory granules along the lateral and basal plasma membranes where contents of the granules were released by exocytosis. Secretory granules in each individual cell were exclusively of one type, either solid or 'haloed', and for all cells observed, the range in granule diameter was 60-120 nm. The cytoplasm varied in density from clear to dark. Lamellar bodies were prominent in the apical and lateral cellular regions and did not exhibit acid phosphatase activity. The basal plasma membrane was smooth adjacent to the basal lamina, whereas in digestive cells the membrane formed a labyrinth. Some endocrine cells reached the midgut lumen and were capped by microvilli; a system of vesicles and tubules extended from beneath the microvilli to the cell body. An estimated 500 endocrine cells were distributed in both the thoracic and abdominal regions of the adult midgut. In one midgut, we classified a sample of endocrine cells according to cytoplasmic density and granule type and size; endocrine cells with certain types of granules had specific distributions within the midgut.     

Morphometric differences in midgut epithelial cells between strains of female Aedes aegypti (L.) (Insecta,Diptera)

Midgut epithelial cells of female Aedes aegypti deriving from 2 different mosquito strains were compared morphometrically. One of the 2 strains was recently isolated from nature (East Africa). The other strain is an old laboratory strain, reared under laboratory conditions for about 30 years. The quantitative morphological comparison demonstrates, that the ultrastructural composition of the midqut epithelial cells corresponds generally in both strains. It can be shown, however, that in the old strain, compared to the new strain, the organelle amount of midgut epithelial cells is significantly reduced during the first 53 days of the mosquito's life. Parallel to decreased relative volumes of organelles and reduced surface densities of membrane systems an increase in lysosome-like structures is observed. These observations are interpreted as deterioration processes, possibly due to the long rearing in the laboratory. Therefore, care should be taken in the selection of an A.aegypti strain for any quantitative morphological or physiological investigation.     

中华硬蜱叮咬不同免疫力新西兰兔后中肠上皮组织的形态动态变化

通过光镜和电镜观察了中华硬蜱Ixodes sinensis叮咬初次和再次感染宿主新西兰兔后不同时间（叮咬后24 h、48 h、72 h以及第 5天、第8天）中肠上皮组织的形态学动态变化。结果显示： 中华硬蜱叮咬前中肠上皮主要由替代细胞和少量体积较大的消化细胞构成;替代细胞数量多、体积小、呈圆形、胞质染色浅 。中华硬蜱叮咬初次感染宿主后,消化细胞随叮咬时间延长而增多增大,微绒毛较密集,排列整齐,胞质内细胞器丰富,各单位膜结构清晰,并出现顶端小管、小泡、大量脂滴和高铁血红素颗粒;近基膜的细胞膜内褶形成发达的基底迷路系统。中华硬蜱叮咬再次感染宿主后,中肠可发生一系列明显的病理变化,中肠基膜出现变薄、松散和断裂现象,消化细胞破裂、空泡化,消化细胞数量减少;消化细胞微绒毛减少、变短、排列不整,线粒体肿大,体嵴减少、变短甚至髓样变,粗面内质网扩张,脂粒及高铁血红素颗粒减少,细胞膜吞饮、吞噬现象减弱,消化细胞内结构紊乱和破坏。该研究结果提示初次叮咬导致了宿主的免疫抗性,再次叮咬后蜱中肠是宿主免疫力的主要作用部位。     

Subcellular localization of the mosquito sterol carrier protein-2 and sterol carrier protein-x

Subcellular distribution of Aedes aegypti sterol carrier protein-2 (AeSCP-2) and AeSCP-x was studied using electron microscopy. In both cultured A. aegypti cells and in the larval midgut, AeSCP-2 was detected mostly in the cytosol, with some labeling mitochondria and nucleus, but not in membranous vesicles. The widespread distribution of AeSCP-2 in the midgut epithelium is consistent with its potential lipid transfer function in all phases of cholesterol absorption. In contrast, AeSCP-x was found mostly in the peroxisome. Differences in the subcellular distribution of AeSCP-2 and AeSCP-x suggest that these two members of the SCP-2 gene family are functionally distinct. Overexpression of AeSCP-2 in A. aegypti cells showed increased localization of AeSCP-2 to cytosol, mitochondria, and nucleus. This is the first report on the nuclear distribution of an SCP. Overexpression of AeSCP-2 resulted in increased cholesterol incorporation in cells, suggesting that AeSCP-2 enhances cholesterol uptake.     

Morphometric analysis of Rhodnius prolixus Stal (Hemiptera:Reduviidae) midgut cells during blood digestion

Post-feeding ultrastructural modifications to the midgut cells of Rhodnius prolixus are quantified using morphometry. Changes in relative and absolute volumes and/or surface areas are demonstrated for the whole cells, nuclei, mitochondria, rough endoplasmic reticulum, lysosomes, Golgi apparatus, storage vesicles, glycogen, microvilli, and basal labyrinth, before and during blood digestion. These parameters are separately determined for cells from each of the three midgut regions, and are correlated against previously published cycles of digestive enzyme activities. The results support the proposed division of the midgut of R. prolixus into three functional regions: the anterior midgut or crop is the site of water transport immediately after feeding, and of lipid and glycogen storage. No protein digestion occurs in this region. The anterior intestine is the site of most proteinase synthesis and secretion, although limited absorption and nutrient storage also occurs. The posterior intestine is responsible for some secretory activity, but is also implicated as the most important region for absorption of digested nutrients and for carbohydrate absorption and storage.     

Ultrastructure and morphology of midgut visceral muscle in early pupal Aedes aegypti mosquitoes

These studies focus on the pupal Aedes aegypti midgut muscularis for the first 26 h following larval-pupal transition. The midgut muscularis of Ae. aegypti pupae during this first half of the pupal stadium is a grid of both circularly and longitudinally oriented muscle bands, arranged in a manner resembling that of the larvae. While many muscle bands exhibit signs of degeneration during the time period studied, not all bands degrade, nor is this degradation simultaneous. Band deterioration involves destruction of internal elements while the muscle fiber plasma membrane remains intact. Deterioration of contractile elements may involve proteosome-like structures and associated enzymes. Many features of the larval muscularis including cruciform cells, bifurcating circular bands, and bifurcating longitudinal bands of muscle are retained during the time period investigated. Neuromuscular junctions along some muscle bands are retained through at least 16 h into the pupal stadium. The selective nature of muscle fiber degradation, coupled with the retention of larval features and neural input, may allow for limited functionality of the muscularis during metamorphosis. Evidence of sexual dimorphism in the midgut muscularis of male and female Ae. aegypti pupae was not observed during the time period studied.     

Functional ultrastructure of the midgut of the fire ant Solenopsis saevissima Forel 1904 (Formicidae: Myrmicinae)

Solenopsis saevissima has a midgut composed of columnar, regenerative, and goblet cells. The midgut epithelium was covered by a basal lamina. Outside the basal lamina, layers of inner oblique, circular, and outer longitudinal muscles were present. Columnar cells showed a basal plasma membrane containing numerous folds, mitochondria, and the nucleus. Rough endoplasmic reticulum, Golgi bodies, membrane bounded vacuoles, and spherocrystals were found in this region. The apical plasma membrane was constituted by microvilli, which were above a region rich in mitochondria. Regenerative cells were found in groups lying by the basal lamina. Goblet cells were associated with an ion-transporting mechanism between the haemolymph and the midgut epithelium. These cells were lying by the midgut lumen and large microvilli were evident, but the cytoplasmic features were similar to the columnar cells.     

Sex dimorphism in the thyroid gland. IV. Cytologic aspects of sex dimorphism in the rat thyroid gland

This study was designed to explain whether the sex-dependent differences in the structure of the thyroid gland of adult male and female rats depend on quantitative or qualitative changes in the thyroid follicular cells. Absolute thyroid gland weight was similar in male and female rats, but its relative weight was markedly higher in females however. Volume fractions of epithelium and stroma were higher and that of colloid lower in male than in female rats and the epithelium/colloid ratio was higher in the males. Also absolute the volumes (in mm3) of epithelium and stroma were higher in the males; the thyroid gland of females contained more colloid. The average volume of a thyroid follicular cell, estimated by stereology, was higher in males than in females, although the thyroid gland contained similar numbers of follicular cells in both sexes. Also, thyroid glands from both male and female rats contained a similar DNA quantity. Results of the present study show that the sex dimorphism in the rat thyroid depends upon a difference in the mean volume of thyroid follicular cells, with males having larger cells than females. However, in both sexes the thyroid gland contains a similar quantity of these cells.     

Ultrastructure of the midgut and blood meal digestion in the adult tickDermacentor variabilis

Digestive cells in the midgut of male and femaleDermacentor variabilis (Say) took up the blood meal in coated vesicles and smooth flask-shaped vesicles, and deposited it in endosomes which were digested via heterophagy. Iron was concentrated in residual bodies.Digestion occurred in three distinct phases in mated females: (1) continuous digestion (initiated by feeding) occurred during slow engorgement; (2) reduced digestion (initiated by mating) occurred in mated females during the period of rapid engorgement; (3) a second continuous digestion phase (initiated by detachment from the host) occurred throughout the post-feeding periods of preoviposition and oviposition.It is proposed that the stem cells in the midguts of unfed females were progenitors of digestive, replacement, and presumed vitellogenic cells in midguts of mated feeding females. Digestive cells were present in all three digestion phases. Only during the first continuous digestion phase did digestive cells fill up with residual bodies, rupture and slough into the lumen, or did whole cells slough into the lumen. During the other two digestion phases no sloughing of digestive cells was observed. At the end of oviposition the digestive cells were filled with residual bodies. Replacement cells were present only during the first continuous-digestion phase. Presumed vitellogenic cells were present only during the reduced-digestion phase and during the second continuous-digestion phase. Stem cells in unfed males developed only into digestive cells in feeding males. Fed males and fed unmated females had only the first continuous-digestion phase. After being hand-detached from the host, unmated 13-day-fed females went through cellular changes associated with the reduced-digestion phase and second continuous-digestion phase of fed mated females, then began ovipositing. Maximum development of the basal labyrinth system and lateral spaces matched the known time of maximum water and ion movement across the midgut epithelia.Spectrophotometric analyses of lumen contents and midgut cells, sampled after detachment from the host, showed that concentrations of protein and hemoglobin at day 1 post-detachment decreased by one-half at the beginning of oviposition, while hematin increased about twofold by the end of oviposition. This supported the idea of the presence of a second continuous-digestion phase.     

Ultrastructure and Immunofluorescence of the midgut of Bombus morio (Hymenoptera: Apidae: Bombini)

Bumblebees are widely distributed across the world and have great economic and ecological importance as pollinators in the forest as well as in agriculture. The insect midgut consists of three cell types, which play various important roles in digestion, absorption, and hormone production. The present study characterized the anterior and posterior midgut regions of the bumblebee, Bombus morio. The digestive, regenerative and endocrine cells in the midgut showed regional differences in their number, nuclear size, as well as the size of the striated border. Ultrastructurally, the digestive cells contained many mitochondria and long microvilli; however, in the anterior midgut region, these cells showed dilated basal labyrinths with a few openings for the hemocoel, whereas the labyrinths of the basal posterior region remained inverse characteristics. Thus, the characterization of the midgut of B. morio supported an ecto-endoperitrophic circulation, contributing to a better understanding of the digestive process in this bee.     

FMRFamide- and pancreatic polypeptide-like immunoreactivity of endocrine cells in the midgut of a mosquito

Immunocytochemical surveys of midguts from female mosquitoes, Aedes aegypti, reveal that half of the estimated 500 endocrine cells in a midgut contain a substance recognized by antisera to bovine pancreatic polypeptide and a molluscan peptide, FMRFamide (phenylalanine-methionine-arginine-phenylalanine-amide). With light microscopy the cells resemble an endocrine type because of their basal position in the epithelium, conical shape, and, in some instances, apical extensions to the lumen. At the ultrastructural level, the immunoreactive substance is contained specifically within the secretory granules of such cells. Immunoreactive cells are distributed exclusively in the midgut region where blood is stored, and ingestion of vertebrate blood reduces the number of such cells and the intensity of reaction in others. These two facts suggest that a blood meal stimulates release of the immunoreactive substance from the cells. Since the immunocytochemical localization is supplemented by a demonstrated secretory response, the cells are considered to be peptidergic endocrine cells.     

THE BASE OF THE PROXIMAL CONVOLUTED TUBULE CELLS OF RAT KIDNEY

The three dimensional arrangement of the compartments on the base of proximal convoluted tubule cells of rat kidney is described. An extracellular basal labyrinth is found to be enclosed by these compartments. The compartments with their mitochondria and the basal labyrinth are regarded as forming a functional unit. It is supposed that this basal unit serves for excretion of reabsorbed fluid from the cell into the labyrinth and for the development of hydrostatic pressure in the labyrinth to overcome the capillary pressure and to pass the reabsorbed fluid into the blood stream.     

In vivo binding of the Cry11Bb toxin of Bacillus thuringiensis subsp. medellin to the midgut of mosquito larvae (Diptera: Culicidae)

Bacillus thuringiensis subsp. medellin produces numerous proteins among which 94 kDa known as Cry11Bb, has mosquitocidal activity. The mode of action of the Cry11 proteins has been described as similar to those of the Cry1 toxins, nevertheless, the mechanism of action is still not clear. In this study we investigated the in vivo binding of the Cry11Bb toxin to the midgut of the insect species Anopheles albimanus, Aedes aegypti, and Culex quinquefasciatus by immunohistochemical analysis. Spodoptera frugiperda was included as negative control. The Cry11Bb protein was detected on the apical microvilli of the midgut epithelial cells, mostly on the posterior midgut and gastric caeca of the three mosquito species. Additionally, the toxin was detected in the Malpighian tubules of An. albimanus, Ae. aegypti, Cx. quinquefasciatus, and in the basal membrane of the epithelial cells of Ae. aegypti midgut. No toxin accumulation was observed in the peritrophic membrane of any of the mosquito species studied. These results confirm that the primary site of action of the Cry11 toxins is the apical membrane of the midgut epithelial cells of mosquito larvae.     

Ultrastructural studies and Na+,K+-ATPase immunolocalization in the antennal urinary glands of the lobster Homarus gammarus (Crustacea, Decapoda).

Unlike in crustacean freshwater species, the structure and ultrastructure of the excretory antennal gland is poorly documented in marine species. The general organization and ultrastructure of the cells and the localization of Na(+),K(+)-ATPase were examined in the antennal gland of the adult lobster Homarus gammarus. Each gland is composed of a centrally located coelomosac surrounded ventrally by a labyrinth divided into two parts (I and II) and dorsally by a voluminous bladder. There is no differentiated nephridal tubule between them. The labyrinth and bladder cells have in common a number of ultrastructural cytological features, including basal membrane infoldings associated with mitochondria, apical microvilli, and cytoplasmic extrusions, and a cytoplasm packed with numerous vacuoles, vesicles, lysosome-like bodies, and swollen mitochondria. Each type of cell also presents distinctive characters. Na(+),K(+)-ATPase was detected through immunofluorescence in the basal part of the cells of the labyrinth and in the bladder cells with an increasing immunostaining from labyrinth I to the bladder. No immunoreactivity was detected in the coelomosac. The cells of the labyrinth and of the bladder present morphological and enzymatic features of ionocytes. The antennal glands of the lobster thus possess active ion exchanges capabilities.     

Sexual Dimorphism in the Expression of Mitochondria-Related Genes in Rat Heart at Different Ages

Cardiovascular disease (CVD) is the leading cause of mortality worldwide. Moreover, sex and age are considered major risk factors in the development of CVDs. Mitochondria are vital for normal cardiac function, and regulation of mitochondrial structure and function may impact susceptibility to CVD. To identify potential role of mitochondria in sex-related differences in susceptibility to CVD, we analyzed the basal expression levels of mitochondria-related genes in the hearts of male and female rats. Whole genome expression profiling was performed in the hearts of young (8-week), adult (21-week), and old (78-week) male and female Fischer 344 rats and the expression of 670 unique genes related to various mitochondrial functions was analyzed. A significant (p<0.05) sexual dimorphism in expression levels of 46, 114, and 41 genes was observed in young, adult and old rats, respectively. Gene Ontology analysis revealed the influence of sex on various biological pathways related to cardiac energy metabolism at different ages. The expression of genes involved in fatty acid metabolism was significantly different between the sexes in young and adult rat hearts. Adult male rats also showed higher expression of genes associated with the pyruvate dehydrogenase complex compared to females. In young and adult hearts, sexual dimorphism was not noted in genes encoding oxidative phosphorylation. In old rats, however, a majority of genes involved in oxidative phosphorylation had higher expression in females compared to males. Such basal differences between the sexes in cardiac expression of genes associated with energy metabolism may indicate a likely involvement of mitochondria in susceptibility to CVDs. In addition, female rats showed lower expression levels of apoptotic genes in hearts compared to males at all ages, which may have implications for better preservation of cardiac mass in females than in males.     

Morphological and enzymatic analysis of the midgut of Anopheles darlingi during blood digestion

The midgut of adult female Anopheles darlingi is comprised of narrow anterior and dilated posterior regions, with a single layered epithelium composed by cuboidal digestive cells. Densely packed apical microvilli and an intricate basal labyrinth characterize each cell pole. Before blood feeding, apical cytoplasm contains numerous round granules and whorled profiles of rough endoplasmic reticulum. Engorgement causes a great distension of midgut. This provokes the flattening of digestive cells and their nuclei. Simultaneously, apical granules disappear, the whorls of endoplasmic reticulum disassemble and 3h post bloodmeal (PBM), nucleoli enlarge manyfold. An intense absorptive process takes place during the first 24 h PBM, with the formation of large glycogen inclusions, which persist after the end of the digestive process. Endoproteases activities are induced after bloodmeal and attain their maximum values between 10 and 36 h PBM. At least two different aminopeptidases seem to participate in the digestive process, with their maximum activity values at 36 and 48 h PBM, respectively. Coarse electrondense aggregates, possibly debris from digested erythrocytes, begin to appear on the luminal face of the peritrophic membrane from 18 h PBM and persist during all the digestive process, and are excreted at its end. We suggest that these aggregates could contain some kind of insoluble form of haem, in order of neutralize its toxicity.