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Glycosaminoglycan composition of the large freshwater mollusc bivalve Anodonta anodonta.

In this paper, glycosaminoglycans from the body of the large freshwater mollusc bivalve Anodonta anodonta were recovered at about 0.6 mg/g of dry tissue, composed of chondroitin sulfate (approximately 38%), nonsulfated chondroitin (about 21%), and heparin (41%). This last polysaccharide was found to consist of a large percentage (approximately 88%) of a fast-moving species possessing a lower molecular mass and sulfate group amount and about 12% of a more sulfated, slow-moving component having a greater molecular mass. The chondroitin sulfate was composed of approximately 28% of the 6-sulfated disaccharide, 46% of the 4-sulfated disaccharide, and about 26% of the nonsulfated disaccharide, with a charge density value of 0.74. Heparin was subjected to the oligosaccharide mapping after treatment with heparinase and then separation of the resulting unsaturated oligosaccharides by SAX-HPLC. A heparin sample from Anodonta anodonta showed a degree of sulfation similar to that of bovine mucosal heparin because of the presence of approximately the same mol % of the trisulfated disaccharide (DeltaUA2S(1-->4)-alpha-D-GlcN2S6S), a slight modification of the other oligosaccharides, and a significant increase of the disaccharide bearing the sulfate group in position 3 of the N-sulfoglucosamine 6-sulfate (-->4)-beta-D-GlcA(1-->4)-alpha-D-GlcN2S3S6S(1-->) part of the ATIII-binding region. However, the anticoagulant activity of mollusc heparin was quite similar to that of pharmaceutical grade heparin. The data obtained again emphasize the heterogeneity of GAGs from molluscs.

Animals↗

[Seasonal changes in myocardial sensitivity to acetylcholine in the molluscs, Anodonta complanata and Anodonta piscinalis].

Seasonal changes in ACh sensitivity have been studied on isolated cardiac muscle of the bivalve molluscs together with ATP content of the hemolymph and the heart. The lowest ACh sensitivity of the heart was found in August and October (KACh in August was equal to 44.0.10(-6) M). Maximum sensitivity was observed in March and at the end of August--the beginning of September (KACh in March was equal to 1.27 X X 10(-6) M); at this period of year, ATP content of the hemolymph decreases approximately 100-fold, the release of ATP from the heart in the presence of ACh being also decreased. Maximum level of ATP in cardiac muscle was found in July. It was demonstrated that most significant changes in ACh sensitivity take place during spawning, especially when sex cells are transported from the gonads into the gills, as well as during the release of glochidia from the latter. Possible causes of seasonal changes in ACh sensitivity of cardiac muscle and the role of ATP in this process are discussed.

Acetylcholine↗

Genetic diversity and divergence among freshwater mussel (Anodonta) populations in the Bonneville Basin of Utah.

Populations of the freshwater mussel genus Anodonta appear to be in a state of rapid decline in western North America, following a trend that unfortunately seems to be prevalent among these animals (Mollusca: Unionoida). Here we describe the patterns of molecular divergence and diversity among Anodonta populations in the Bonneville Basin, a large sub-basin of the Great Basin in western North America. Using amplified fragment length polymorphism (AFLP) analysis, we found a striking lack of nuclear diversity within some of these populations, along with a high degree of structuring among populations (FST = 0.61), suggesting post-Pleistocene isolation, due either to a long-term loss of hydrologic connectivity among populations or to more recent fish introductions. We also found evidence of recent hybridization in one of these populations, possibly mediated by fish-stocking practices. Using mitochondrial sequence data, we compared the Bonneville Basin populations to Anodonta in several other drainages in western North America. We found a general lack of resolution in these phylogenetic reconstructions, although there was a tendency for the Bonneville Basin Anodonta (tentatively A. californiensis) to cluster with A. oregonensis from the adjacent Lahontan Basin in Nevada. We recommend further investigation of anthropogenic factors that may be contributing to the decline of western Anodonta and a broad-scale analysis and synthesis of genetic and morphological variation among Anodonta in western North America.

Animals↗

The fine structure and innervation of gill lamellae in Anodonta.

Gill lamellae of a bivalve Anodonta woodiana lauta (v. Martens) were observed by electron microscopy. The Anodonta gill wall consists of a single layer of epithelial cells, its basal lamina and the underlying connective tissue layer. It was confirmed that there is no true endothelium in the vessels and that the connective tissue layer of the vessel wall is therefore in direct contact with the blood. Cells of a specific type referred to as "trabecular cells" lie in the blood lacunae. These cells closely resemble the pillar cells of fish gills, but show certain fundamental differences. Characteristic features of the trabecular cells are (1) an elongated cell body which lies across the vascular lumen and attaches to the vessel wall by means of the tips of their long processes, (2) two types of myofilaments (thick and thin) in the cytoplasm, (3) external dense plaques at the cell surface which are associated with the insertion of myofilaments into the cell membrane, (4) direct contact between the cell surface and the blood except at the regions where the cell is covered by external plaques and connective tissue fibrils. These facts suggest that the Anodonta trabecular cell is not analogous with the pillar cell of fish gills but rather with muscle cells which show a specific morphological modification and a peculiar relationship to the vessel wall due to the absence of the endothelium. These cells are assumed to regulate blood flow within the gill vessels. As to the permeability of the wall of Anodonta gill vessels, junctional complex consisting of an intermediate and a septate junction between adjacent gill epithelial cells probably plays the main role as a barrier between the blood and the surrounding water. The basal lamina underlying the gill epithelium is assumed to act as a coarse permeability barrier. Numerous nerve endings of unknown function are observed in the gill epitheliu. It is strongly suggested, however, that they are associated with the additional function of the Anodonta gill lamellae as a food-sorting device.

Animals↗

[Filtering capacity of Anodonta woodiana and its feeding selectivity on phytoplankton].

Lake eutrophication associated with algal bloom has become a major environmental challenge in China. Filter feeding organisms, e.g., silver carp, bighead carp, and zebra mussel, have been widely used to manipulate water environment through their top-down effects. The bivalve Anodonta woodiana is widely distributed across China, and has been used for bio-manipulation of some water bodies. In situ experiments were performed in a bay of Lake Dianchi and its surrounding ponds to study the filtering capacity and the filter feeding rate of Anodonta woodiana in these aguatic habitats, and to determine the controlling factors. The results showed that there was a relationship between filtration rate and total suspended substances measured in laboratory during August and September 2001 when the water temperature was 20 +/- 2 degrees C, which was expressed as y = 4 380.41 e(- 0.0184x) (R2 = 0.621, P < 0.05) based on at least squares analyses. The filtration rate was strongly related to the body weight by measuring in laboratory over 24 hours during August 2001 when the water temperature was 20 +/- 2 C. At least squares analyses gave the equation y = 2 904.16 + 12 076.24e(-0.0034x) (R2 = 0.9825, P = < 0.05). A slight variation of filtration rate was found over 24 hours, which could be explained by the bivalves' adaptation to the experimental circumstances rather than the daily change of its filtration feeding. In general, the filtration capacity of natural Anodonta woodiana population in the water bodies was very limited, as a result of its small population size. No significant selective feeding on algae was found by contrasting the relative abundance of algae in the habitats and the guts of Anodonta woodiana. The feeding selectivity on algae was neither correlated with the size of algae, nor with the initial abundance of algae in water column.

Animals↗

Accumulation and depuration of the cyanobacterial toxin cylindrospermopsin in the freshwater mussel Anodonta cygnea.

Cylindrospermopsin (CYN) is a toxic alkaloid produced by several genera of freshwater cyanobacteria. This compound has been implicated in outbreaks of human sickness and the death of domestic and wild animals. Given that several of the cyanobacterial genera known to produce CYN are common components of the phytoplankton of freshwaters including aquaculture facilities, we studied the accumulation of CYN in the freshwater mussel (swan mussel) Anodonta cygnea. Anodonta were exposed to CYN-producing cultures of the cyanobacterium Cylindrospermopsis raciborskii for 16 days and were found to accumulate the toxin to concentrations up to 2.52 microg g tissue dry weight(-1). There was considerable variation in the concentrations of CYN detected in different parts of the body. At the end of a 2-week accumulation period the distribution of CYN in the body of Anodonta was as follows: haemolymph (68.1%), viscera (23.3%), foot and gonad (7.7%) and mantle (0.9%). No CYN was detected in the gills or adductor muscle of any animals. Following a 2-week depuration period, approximately 50% of the toxin remained in the tissues. Based on the recently derived guideline value for CYN in human drinking water (1 microg l(-1)) and the concentrations of this compound in animal tissues reported here, there is a clear need for the increased monitoring of this compound in organisms grown for human and animal consumption.

Alkaloids↗

[Certain biochemical characteristics of an insulin-like substance from the bivalve mollusk Anodonta cygnea].

An insulin-like substance (ILS) was isolated from the visceral organs of the bivalve mollusc Anodonta cygnea by chromatography on a sulfocationite CU-23 and purified by reverse phase liquid chromatography. ILS was shown to be made up to several fractions with Mr ranging from 9 to 20 kDa which have identical amino acid composition but different hydrophobicity and N-terminal amino acids. It was supposed that the heterogeneity of ILS fractions is due to its genetical or posttranslational polymorphism. ILS has a low (0.02%) affinity for the mammalian insulin receptor and a low immune affinity for mammalian insulin and possesses a mitogenic activity which is commensurate with that of the epidermal growth factor. The data obtained suggest that Anodonta cygnea ILS represents a separate branch of a relatively ancient family of insulin-like hormones and growth factors responsible for metabolism and proliferation of invertebrate tissues.

Amino Acids↗

Light and Electron microscopic autoradiography of 3H-dopamine uptake in the ganglia of the freshwater mussel, Anodonta cygnea L.

In the central nervous system of the freshwater mussel, Anodonta cygnea, uptake of 3H-dopamine has been investigated by light and electron microscopic autoradiography following in vitro incubation and topical application, respectively. The autoradiographic reaction observed over the ganglia can be taken as a morphological evidence of the existence of a specific uptake mechanism for dopamine. It has been found that labeled dopamine was taken up by the axon profiles and terminals, while perikarya remained free of activity, indicating a basic difference between the perkaryal and axonal membrane in respect of the 3H-dopamine uptake. According to the electron microscopic autoradiography, labeled terminals and axon profiles contained both normal and accentric dense-core vesicles and they proved to be similar to those seen after 3H-serotonin in vitro incubation. Therefore, it seems possible that certain Anodonta central neurons contain both dopamine and serotonin.

Animals↗

[Structural characteristics of lysine-rich histone from the sperm of a mollusk Anodonta piscinalis].

Sperm of freshwater bivalve mollusk Anodonta piscinalis was found to contain two fractions of lysine-rich histone: somatic histone H1 and sperm-specific protamine-like histone, named Hp. A detailed analysis of H1 and Hp structure was carried out by means of N-bromosuccinimide, chymotrypsin and pepsin cleavage followed by determination of the lysine residue number, positive charge and molecular length of obtained fragments by the method of incomplete succinylation. It has been shown, that Anodonta histone H1, like the avian histone H5, contains 3 tyrosine residues in the central hydrophobic domain of the molecule. Histone Hp contains 5 tyrosine residues, 3 of which are localized in the hydrophobic domain, while the rest two--in the COOH-terminal part of the molecule, characterized by a strong positive charge. Such unusual disposition of tyrosine residues in the lysine-rich histone has been found for the first time. All the regions of histone Hp molecule contain a great number of arginine residues. The only phenylalanine residue is localised approximately in the middle of the polypeptide chain for both H1 and Hp molecules. On the basis of structure homology between histones H1 and Hp the origin of Hp from H1 in the course of evolution is proposed.

Amino Acid Sequence↗

Burrowing behaviour affects Paraergasilus rylovi abundance in Anodonta piscinalis.

Burrowing depth may affect predation rate, feeding ability and reproduction in bivalve clams. We studied the effect of burrowing depth on the abundance of the ergasilid Paraergasilus rylovi in the freshwater bivalve clam Anodonta piscinalis. We transplanted uninfected clams to a lake where they were allowed to choose their preferred burrowing depth, and were exposed naturally to copepodids of the parasite. There was a significant positive correlation between proportionate burrowing depth (PBD) and the abundance of P. rylovi at the end of the 17-day experiment, the deeper-burrowed clams harbouring more P. rylovi. Original PBD (0%, 50%, 100%) did not influence the final PBD or parasite abundance. Clam length affected PBD, smaller clams burrowing deeper, but it did not affect parasite abundance. Infected experimental clams and naturally-burrowed uninfected clams, both originating from the same lake, did not differ in their mean PBD. This indicated that burrowing of the experimental clams affected parasitism rather than the parasites altering burrowing of the clams. In line with the experimental result, we observed a significant positive correlation between PBD and the abundance of P. rylovi also among clams collected from 2 natural A. piscinalis populations.

Animals↗

Pleiotropic action of insulin-like peptides of mollusk, Anodonta cygnea.

Insulin-related peptides (IRPs) from ganglions of mollusk, Anodonta cygnea, were purified and characterized (IRP1-IRP13) using insulin and insulin-like growth factor-I (IGF-I) radioreceptor test systems. The IRPs were able to bind to insulin and IGF-I receptors. Dose-dependent curve slopes indicated that most IRPs bind with higher affinity to IGF-I receptors than to insulin receptors. The IRP regulatory action on the activity of the adenylyl cyclase signal system showed that these peptides stimulated adenylyl cyclase and GTP-binding activity of G-proteins to the same extent as insulin and IGF-I. The data obtained suggest polyfunctional IRP action that apparently is determined by the molecular structure of individual isoforms.

Animals↗

Sensitivity of adenylyl cyclase signaling system of the mollusk Anodonta cygnea ganglions to serotonin and adrenergic agonists.

For the first time, the adenylyl cyclase system (ACS) sensitive to biogenic amines in the mollusk Anodonta cygnea ganglions was revealed and characterized. Serotonin and isoproterenol stimulated AC activity and GTP-binding activity of heterotrimeric G-proteins. The AC-stimulating action of serotonin and isoproterenol was blocked by cyproheptadine and adrenergic antagonists, respectively. Using synthetic C-terminal peptides of G-protein alpha-subunit, it was shown that the biogenic amines realized their action on the ACS through different G-protein types: serotonin and isoproterenol activate G(s)-protein, while adrenaline preferably activates G(i)-protein.

Adenylyl Cyclases↗

Paracellular pathway in the shell epithelium of Anodonta cygnea.

Ultrastructural study of cell-cell connections in the outer mantle epithelium (OME) on high-pressure-frozen specimens revealed zonula adherens, septate junctions and gap junctions in Anodonta cygnea. In order to evaluate the permeability of the paracellular pathway, the OME was incubated under gradients of lanthanum and calcium. After lanthanum incubation (4 mM) from the basal side, the septate junctions were penetrated completely by this tracer. When applied from the apical side, lanthanum deposits were located similarly over the entire length of the septate junctions up to the first dilatations of the intercellular space. Calcium deposits were also present in paracellular areas only when OME had been incubated simultaneously with calcium (6 mM) and lanthanum (4 mM) gradients. Lanthanum and calcium deposits were detected with ESI (Electron Spectroscopic Imaging) and identified with EELS (Electron Energy Loss Spectroscopy). On the other hand, electrophysiological observations showed a 48% reduction of conductance when the OME was bathed on both sides with solutions containing lanthanum (4 mM) and calcium (6 mM), compared to bathing with lanthanum-free solution (control). The conductance reduction was 52% when calcium was removed from the control solution. Supported by morphological and physiological evidence, it appears that, under in vivo conditions, calcium ions may diffuse paracellularly from the haemolymph towards the extrapallial fluid and vice-versa across the septate junctions in the OME of A. cygnea. Permeability of the septate junctions depended proportionally on the calcium concentration in fluids.

Adherens Junctions↗

Short- and long-term effects of deltamethrin on filtering activity of freshwater mussel (Anodonta cygnea L.).

Bioindicator methods were used to study the effects of the pyrethroid insecticide deltamethrin on the filtering behavior of the freshwater mussel Anodonta cygnea L in the laboratory, during short-term (30 min) and long-term (1 week) exposure. In the course of the short-term treatment, the water flow through the outflow syphon was monitored. It was found that 1 and 5 micrograms/liter deltamethrin caused an increase, whereas from 10 to 50 micrograms/liter deltamethrin resulted in a depression, in the open time of the outflow syphon and the water outflow. The inhibitory response was concentration dependent. In the long-term experiments, the valve movement representing the activity of the adductor muscles was recorded. At 1 microgram/liter, deltamethrin had no effect on the active period, but reduced the rest time, causing an increase in the filtration; all higher concentrations (10-50 micrograms/liter) caused inhibition of the filtration activity by reducing the active periods and lengthening the rest periods. The active periods were shortened on increase in deltamethrin concentration, the reduction being 80% of the control at 50 micrograms/liter. Lengthening of the rest periods led to a maximum (207% as compared with the control) at 15 micrograms/liter, with a subsequent decline to levels close to the control at 30 and 50 micrograms/liter. Nevertheless, due to the strong reduction of the active periods, the filtering activity remained depressed at higher concentrations. The results provide evidence that deltamethrin pollution may have an adverse effect on the functioning of the bivalve community in the aquatic environment, which should be considered when this chemical is used in agricultural areas near aquatic ecosystems.

Animals↗