Hepatic ultrastructural specialization in Antarctic fishes

Summary Compared with those of other vertebrate animals, the livers of Antarctic fishes have a unique type of perisinusoidal (Ito) cell. These cells were studied in 9 species with emphasis on Dissostichus mawsoni. Perisinusoidal cells are found in large numbers throughout the liver, have long cytoplasmic arms and, in Dissostichus, contain numerous lipid droplets. The extensive rough endoplasmic reticulum and prominent nucleolus are ultrastructural characteristics indicating that these cells are engaged in protein synthesis. An evolutionary specialization, perisinusoidal cells may be partially responsible for the elevated levels of protein synthesis characteristic of fishes in the Antarctic marine environment.     

Intralobular heterogeneity of perisinusoidal stellate cells in porcine liver

The aim of the present investigation was to elucidate the intralobular heterogeneity of the perisinusoidal stellate cells (fat-storing cells, lipocytes) in the porcine liver. Their three-dimensional structure, desmin immunoreactivity and vitamin-A storage were studied by use of the Golgi silver, immunocytochemical and gold chloride methods. In order to locate the stellate cells, the hepatic lobules were divided into 10 zones. The stellate cells were readily identified in Golgi preparations by their striking dendritic appearance with branching processes encompassing the sinusoids. The stellate cells in the centrolobular zones were conspicuously dendritic with longer processes in conspicuously dendritic with longer processes in comparison to those emitted by periportal elements. Such arborizations were studded with numerous thorn-like microprojections. Desmin immunoreaction in the periportal zones was stronger than that in the centrolobular zones. Vitamin-A storage in the stellate cells was well developed in zones 2–4, but reduced gradually toward the central region. The perisinusoidal etellate cells display marked heterogeneity in morphology and function based on their zonal location in the hepatic lobule.     

Intercellular communication among liver cells in the perisinusoidal space of the injured liver: Pathophysiology and therapeutic directions

Hepatic stellate cells (HSCs) in the perisinusoidal space are surrounded by hepatocytes, liver sinusoidal endothelial cells, Kupffer cells, and other resident immune cells. In the normal liver, HSCs communicate with these cells to maintain normal liver functions. However, after chronic liver injury, injured hepatocytes release several proinflammatory mediators, reactive oxygen species, and damage-associated molecular patterns into the perisinusoidal space. Consequently, such alteration activates quiescent HSCs to acquire a myofibroblast-like phenotype and express high amounts of transforming growth factor-β1, angiopoietins, vascular endothelial growth factors, interleukins 6 and 8, fibril forming collagens, laminin, and E-cadherin. These phenotypic and functional transdifferentiation lead to hepatic fibrosis with a typical abnormal extracellular matrix synthesis and disorganization of the perisinusoidal space of the injured liver. Those changes provide a favorable environment that regulates tumor cell proliferation, migration, adhesion, and survival in the perisinusoidal space. Such tumor cells by releasing transforming growth factor-β1 and other cytokines, will, in turn, activate and deeply interact with HSCs via a bidirectional loop. Furthermore, hepatocellular carcinoma-derived mediators convert HSCs and macrophages into protumorigenic cell populations. Thus, the perisinusoidal space serves as a critical hub for activating HSCs and their interactions with other cell types, which cause a variety of liver diseases such as hepatic inflammation, fibrosis, cirrhosis, and their complications, such as portal hypertension and hepatocellular carcinoma. Therefore, targeting the crosstalk between activated HSCs and tumor cells/immune cells in the tumor microenvironment may also support a promising therapeutic strategy.     

A study on cell lineage,especially the germ cell line,in embryos of the teleost fish,Barbus conchonius

Summary Lucifer Yellow-Dextran labelling of lower layer cells (LLC), sometimes together with upper layer cells (ULC), of the 64-cellBarbus conchonius embryo resulted in labelled primordial germ cells (PGCs) at 12 h after fertilization (a.f.) in about 25% of cases. The presence of labelled PGCs was independent of the location of the injected blastomere with respect to the later orientation of the embryonic axis. After injection of an ULC alone, however, labelled PGCs were never found. Also, the distribution of labelled somatic cells differed between the ULC- and LLC-injected embryos. When we found fluorescent PGCs, only a few of them were labelled, suggesting that either a single predecessor exists earlier than the 64-cell stage or that the formation of germ cells is a polyclonal process. Tracing the fluorescent cells at successive stages of development shows an extensive mixing with unlabelled cells during the epiboly stage, which might well be the cause of partly unpredictable cell lineages. The chance of being committed to a specific fate is different for the ULC and LLC descendants. This might be due to relatively limited cell mixing between these two cell populations.     

A hepatic protein modulates glucokinase activity in fish and avian liver: a comparative study

Glucokinase (GCK) is a key enzyme involved in hepatic glucose metabolism as well as in glucose homeostasis regulation. In mammals, GCK is regulated in vivo by a regulatory protein (GCKR) through a nucleus-to-cytoplasm translocation enhanced by fructose 1-phosphate and counteracted by fructose 6-phosphate. There were no previous evidences in literature regarding the presence of GCKR in livers of other vertebrates like fish and bird. Accordingly, in the present study we assessed GCKR presence in chicken, trout, carp, and goldfish hepatic homogenates. The results obtained demonstrate for the first time the presence of a GCKR-like protein in the liver of those species, with molecular weight (68 kDa) and biochemical properties similar to those described in mammals. Several of the biochemical properties of rainbow trout GCKR-like protein were closer to the mammalian model whereas those of chicken protein were specific. We also compared the presence and properties of GCKR-like protein in livers of different teleost species that exhibit different tolerances to glucose such as rainbow trout (intolerant) and goldfish and common carp (tolerant). The results showed that the most powerful GCKR-like protein was found in the most intolerant species, whereas the inhibition exerted by GCKR-like protein in tolerant species was closer to chicken than to rat. Furthermore, the response of GCKR-like protein in liver of rainbow trout fed with a diet rich in carbohydrates was compared with the rat model under extreme glycemic conditions. We found that despite trout GCKR-like protein was less active and expressed than in rat, the response against glycemic changes took place in the same direction, and the ratio GCKR-like protein:GCK was affected in a similar way.     

Labyrinth cells,a new cell type in vertebrate olfactory organs

Summary Light microscopy and transverse electron microscopy has been employed to study the olfactory organs in 82 specimens of freshwater adapted young and homing adult Baltic sea trout Salmo trutta trutta L. In both sensory and indifferent epithelium the olfactory mucosa has scattered cells of a type that has not been described in any olfactory organ before. They are called labyrinth cells and are characterized by an extensive, turtuous, interconnected tubular system of smooth endoplasmatic reticulum intimately connected with numerous mitochondria. This cell type is similar to chloride and other cells which probably are involved in electrolyte transport in fish gills and pseudobranch, the rectal gland in elasmobranchs and the nasal gland in reptiles and birds. It is suggested that the olfactory organ in fish is serially homologous with the pseudobranch.Thanks are due to Prof. Dr. Gunnar Bloom, Section of Histology, University of Umeå for interesting discussions. The author also wish to acknowledge the technical facilities and assistance in the use of the electron microscope to Miss Karin Ekström and Miss Marianne Borg. The research was supported by grant 2389-11, 13 and 15 from the Swedish Natural Science Research Council.     

Plural light chains in a single plasma cell of a monoclonal gammopathy undetermined significance case: An ultrastructural study

A 44-year-old man was found to have M-proteins of IgG consisting of kappa- and lambda-chains in serum without lymphadenopathy or splenomegaly. The serum concentrations of IgG, IgA and IgM were within normal limits. Bone marrow examination showed normal cellular marrow containing 6.3% of plasma cells with no abnormal features. No chromosomal abnormality was observed at all. The patient was diagnosed as having monoclonal gammopathy of undetermined significance. The bone marrow plasma cells possessed free kappa- and lambda-chains in Golgi apparatus, rough endoplasmic reticula and cytoplasmic matrices. Plural light chains were simultaneously produced with the same heavy chain in a plasma cell by immunoelectron microscopy. This is the first report in the world of a monoclonal gammopathy of undetermined significance producing plural light chains with the same heavy chain.     

Growth of fish cell lines in glutamine-free media

The glutamine requirement for thein vitro proliferation of fish cells was investigated with cell lines from four different species and three tissues: goldfish skin (GFSk-S1), Chinook salmon embryo (CHSE-214), and raibow trout liver (RTL-W1) and spleen (RTSp-W1). With a supplement of fetal bovine serum, the basal medium, Leibovitz's L-15, without glutamine supported the proliferation of all four cell lines as well, or nearly as well, as L-15 with 2 mM glutamine. This was true over short term assays of two to four weeks and for continuous propagation. CHSE-214 also grew as well with or without 2 mM glutamine in Minimum Essential Medium with fetal bovine serum. However, when the supplement was dialyzed fetal bovine serum, CHSE-214 grew much better in L-15 without glutamine. Therefore, glutamine was not required for growth in L-15, and in fact, was inhibitory in the absence of the dialyzable fraction of serum. By contrast, glutamine appeared to be important for growth in Minimum Essential Medium. When the supplement was dialyzed fetal bovine serum, CHSE-214 grew much better in Minimum Essential Medium with 2 mM glutamine. These results suggest that the glutamine requirement for thein vitro proliferation of fish cells is conditional and depends on the basal medium and serum supplement.Abbreviations BSA bovine serum albumin - CHSE-214 Chinook samon embryo cell line - dFBS dialyzed fetal bovine serum - FBS fetal bovine serum - GFSk-S1 goldfish skin cell line - GS glutamine synthetase - L-15 Leibovitz's L-15 media - L929 mouse fibroblast cell line - MEM minimum essential medium Eagle - PBS phosphate buffered saline - RTL-W1 rainbow trout liver cell line - RTSp-W1 rainbow trout spleen cell line     

An ultrastructural study of cell junctions and the cytoskeleton in epithelial cells of the molluscan integument

Cell junctions and the cytoskeleton of integumental epidermal cells from six bivalves, four gastropods, and two cephalopods were studied by transmission electron microscopy. In all species examined, the junctions in supporting cells presented the following similar pattern: an apical-lateral adhesion belt (occluding junctions were not observed); (b) a lateral complex of septate junctions and smooth septate junctions, with interdigitations between adjacent cells while the gap junctions were not constantly present, and a basal complex with hemidesmosomes, focal contacts, and sometimes basolateral adherent junctions. Desmosomes were never observed. Microfilamentous and microgranular material were present throughout the cells, as bundles of microfilaments within microvilli and the terminal web, within interdigitations, and as cytoplasmic plaques forming part of the adherent junctions, hemidesmosomes, and focal contacts. Bundles of intermediate filaments that originated from basal hemidesmosomes were located close to and oriented parallel with the lateral plasma membrane and terminated within the terminal web. In cells of Aplysia depilans, intermediate filaments converged apically to terminate in hemidesmosome-like structures at the bases of the microvilli. In the cephalopods, hemidesmosomes were never observed and intermediate filaments made direct contact with the basal cell membrane. Some functional interpretations and hypotheses were also discussed.     

An analysis of fish distribution in Kainji lake Nigeria

The distribution of fish in the three major habitats (inshore, surface and bottom) in Kainji Lake was studied using gill-nets in graded fleets each consisting of seven nets with mesh sizes ranging from 5 cm to 18 cm. No significant difference was observed in the annual yields of fish in the three habitats. Three of the nine major families, however, gave higher yields in selected habitats. Characidae and Cichlidae were most abundant in the shore and surface and Bagridae in the shore and bottom. Catches of the Cichlidae in the surface were limited to areas where the mean depth was below 7 meters. Analysis of variance showed no significant difference in the overall yields of fish among sampling areas located within the major strata of the lake. Although a general trend was observed in the seasonal yields of fish, with higher yields during low water, the variations were not significant either for the overall annual yields or among sets. The implications of these findings in relation to the mode of life of the major fish species and the stability of the fish population are discussed.     

Filtration effect of endothelial fenestrations on chylomicron transport in neonatal rat liver sinusoids

Summary The function of endothelial fenestrations in the control of the passage of chylomicrons from the blood to the liver parenchymal cells was studied ultrastructurally in neonatal rats. Measurement of circulating chylomicrons and endothelial fenestrations as well as chylomicrons in the space of Disse at 2–20 h after the onset of suckling revealed a substantial filtration effect of endothelial fenestrations on chylomicrons larger than 250 nm in diameter. Direct uptake of chylomicrons by parenchymal cells was observed to occur by pinocytosis; Kupffer cells endocytosed only very few chylomicrons, and endothelial cells were inactive in this respect.     

An analysis of fish community responses to coral mining in the Maldives

Synopsis Coral mining takes place on shallow reef flats at a number of localities in the Maldives, but not on the adjacent deeper reef slopes. A semi-quantitative census method for fish species abundance and biomass is described. Fish community structure is compared on mined and non-mined reef flats and their adjacent slopes using a variety of univariate, graphical/distributional and multivariate statistical techniques. In general, univariate and graphical distributional methods do not indicate significant differences between mined and non-mined localities with respect to the relative abundances and biomasses of species. Multivariate methods (both classification and ordination), however, indicate very clear-cut effects of mining on the reef flats, and also significant effects on reef slopes adjacent to mined flats. The effect was equally clear using non-quantitative (presence/absence) data. The fish species mainly responsible for the differences between mined and non-mined localities are identified, and the differences are explained in terms of the feeding biology of these species.     

Effects of noradrenaline exposure on rat brown adipocytes in cultures. An ultrastructural study

Adipocytes in intact brown adipose tissue show multivacuolar lipid deposit and mitochondria of 'typical' morphology. Cultured brown adipocytes retain the multivacuolar lipid deposit, while 'typical' mitochondria degenerate and 'atypical' organelles appear instead of the former. Since evidence exists that catecholamines deeply influence brown adipose tissue morphology and function in vivo, we undertook the present ultrastructural investigation to assess whether exposure of cultured brown fat cell to noradrenaline could prevent (or induce regression of) the in vitro morphological modifications of brown adipocytes. Brown adipocytes cultured for 8 h in the presence of noradrenaline (5 X 10(-5) M) had a larger mitochondrial area (i.e. a larger percentage of cytoplasm occupied by non-degenerating mitochondria) in comparison with control cells, as assessed by morphometry; this was due to larger number of mitochondria in noradrenaline-treated cells. Moreover, a number of cells with mitochondria strictly resembling those of the intact tissue were visible in noradrenaline-treated cultured after 8 hr, while 'typical' mitochondria were no longer observed in parallel control cultures. After 5 days of culture without hormone addition, exposure to noradrenaline (5 X 10(-5) M) did not induce quantitative modifications of 'atypical' mitochondria or changes of their ultrastructure up to 12 hr. However, reduction in size of the lipid deposit and activation of both rough endoplasmic reticulum and Golgi apparatus were evident in noradrenaline-treated adipocytes in comparison with non-treated cells.     

An ultrastructural study of oogenesis in a marine triclad

The ultrastructural features of oocyte differentiation were studied in the marine triclad Cercyra hastata. Oocytes at several stages of maturation, each surrounded by follicle cell projections, are present within each of the two ovaries. A pre-vitellogenic and a vitellogenic stage have been detected in the oogenesis of C. hastata. The pre-vitellogenic stage is mainly characterized by an increase in the nuclear and nucleolar volume and activity, and the appearance and development of cortical granule precursors which are elaborated by the Golgi complex. In early phases of the vitellogenic stage, intense delamination and blebbing of the nuclear envelope occurs which probably contributes to an increase in number of cytoplasmic membranes and to transfer of nuclear material to the cytoplasm. The rough endoplasmic reticulum is extensively developed and often assumes a ‘whorl’ array. Several areas of yolk precursor formation appear in the whorls. Numerous 2–5 μm protein yolk globules are subsequently formed which appear surrounded by a double membrane (cisternae of the smooth endoplasmic reticulum) and become randomly distributed throughout the cytoplasm of mature oocytes. The peripheral ooplasm is occupied by a monolayer of electron-dense cortical granules. Finally, the evolutionary significance of the autosynthetic mechanism of yolk production is discussed.     

Effects of exposure to multiple trace metals on biochemical, histological and ultrastructural features of gills of a freshwater fish, Channa punctata Bloch

The trace metals are frequently encountered as mixtures of essential and non-essential elements. Therefore, evaluation of their toxic effects individually does not offer a realistic estimate of their impact on biological processes. We studied effects of a mixture of four essential and toxic metals (Cu, Cd, Fe and Ni) on biochemical and morphological characteristics of the gills of a biomarker freshwater fish Channa punctata (Bloch) using environmentally relevant concentrations. Fish were exposed to metal mixture through tank water for 7, 15 and 30 days. Biochemical studies as well as light microscopy (LM) and scanning electron microscopy (SEM) revealed significant metal exposure-induced alterations in gills. Besides ultastructural changes, activities of antioxidant enzymes such catalase (CAT), glutathione S-transferase (GST) and superoxide dismutase (SOD) were significantly altered in the gills of exposed fish. The reduced glutathione (GSH) was significantly (p<0.001) decreased, while lipid peroxidation (LPO) was significantly (p<0.001) increased. The main alterations in general morphology of fish gills included spiking and fusion of secondary lamellae, formation of club-shaped filaments, and vacuolization and necrosis of filament epithelium in the interlamellar regions. SEM studies showed gradual increase of the density and apical surface area of the chloride cells and transformation of the surface structure of the pavement cells. The results of this study indicate adaptive as well a toxic responses in fish gills exposed to mixture of trace metals. Low concentrations of trace metal appear to compromise the antioxidant defense of gills. Lesions in the gill morphology caused by the effect of low concentrations of trace metals could lead to functional alterations and interference with fundamental processes such as maintenance of osmoregulation, gas exchange and xenobiotic metabolism in the exposed fish populations.