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PubMed2004

Ligand-binding specificity of an invertebrate (Manduca sexta) putative cellular retinoic acid binding protein.

Intracellular lipid-binding proteins (iLBPs) are small cytoplasmic proteins that specifically interact with hydrophobic ligands. Fatty acid-binding proteins (FABPs), cellular retinoic acid-binding proteins (CRABPs) and cellular retinol-binding proteins (CRBPs) belong to the iLBP family. A recently identified insect (Manduca sexta) iLBP has been reported to possibly represent an invertebrate CRABP mimicking the role of CRABPs in vertebrate organisms. The presence in this protein of the characteristic binding triad residues involved in the interaction with ligand carboxylate head groups, a feature pertaining to several FABPs and to CRABPs, and the close phylogenetic relationships with both groups of vertebrate heart-type FABPs and CRBPs/CRABPs, makes it difficult to assign it to either FABPs or CRABPs. However, its negligible interaction with retinoic acid and high affinity (K(d) values in the 10(-8) M range) for fatty acids have been established by means of direct and competitive binding assays. As shown by phylogenetic analysis, the M. sexta iLBP belongs to a wide group of invertebrate iLBPs, which, besides being closely related phylogenetically, share distinctive features, such as the conservation of chemically distinct residues in their amino acid sequences and the ability to bind fatty acids. Our results are in keeping with the lack of cellular retinoid-binding proteins in invertebrates and with their later appearance during the course of chordate evolution.

Amino Acid Sequence↗

PubMed2004

Cloning and functional expression of invertebrate connexins from Halocynthia pyriformis.

Unlike many other ion channels, unrelated gene families encode gap junctions in different animal phyla. Connexin and pannexin genes are found in deuterostomes, while protostomal species use innexin genes. Connexins are often described as vertebrate genes, despite the existence of invertebrate deuterostomes. We have cloned connexin sequences from an invertebrate chordate, Halocynthia pyriformis. Invertebrate connexins shared 25-40% sequence identity with human connexins, had extracellular domains containing six invariant cysteine residues, coding regions that were interrupted by introns, and formed functional channels in vitro. These data show that gap junction channels based on connexins are present in animals that predate vertebrate evolution.

Amino Acid Sequence↗

PubMed2005

Observations on the history of non-insect invertebrate pathology from the perspective of a participant.

This paper discusses diseases of commercially important non-insect invertebrates from both a historical and a personal perspective. Early studies of curious scientists such as Metchnikoff using invertebrate systems led to important discoveries with much wider impact. As commercial culture of oysters, shrimp, and other invertebrates grew, so did studies of their diseases. Here, the impact of pioneering scientists and important new techniques on our understanding of these diseases is described.

Animals↗

PubMed2005

Exposure to 2,4-decadienal negatively impacts upon marine invertebrate larval fitness.

Diatoms liberate volatile, biologically active unsaturated aldehydes following cell damage, which negatively impact upon invertebrate reproductive processes such as fertilization, embryogenesis and larval survival. 2,4-Decadienal is frequently identified among the aldehydes produced and is one of the more biologically active. The majority of studies which have examined the toxic effects of diatom aldehydes to invertebrate reproduction have scored egg production and/or hatching success as indicators of biological impacts. There are very few studies which have dealt specifically with the impacts of diatom-derived aldehydes on larval fitness. Larval stages of the polychaetes Arenicola marina and Nereis virens and the echinoderms Asterias rubens and Psammechinus miliaris exposed to 2,4-decadienal at sub 1 microg ml(-1) concentrations suffered reduced survival over the incubation period (day 1-8 post fertilization) with detectable differences for the polychates at a concentration of 0.005 and 0.01-0.1 microg ml(-1) for the echinoderms. Susceptibility of larval N. virens was investigated using stage specific 24 h exposures at 2,4-decadienal concentrations up to 1.5 microg ml(-1). A clear stage specific effect was found, with earlier larval stages most vulnerable. Nectochaete larvae (9-10 d) showed no reduction in survival at the concentrations assayed. Fluctuating asymmetry (FA), defined as random deviations from perfect bilateral symmetry, was used to analyse fitness of larval P. miliaris exposed to 2,4-decadienal at concentrations of 0.1, 0.5 and 1 microg ml(-1). The degree and frequency of asymmetrical development increased with increasing 2,4-decadienal concentration. Equally, as FA increased larval survival decreased. These results provide further support for the teratogenic nature of 2,4-decadienal and its negative impact on invertebrate larval fitness.

Aldehydes↗

PubMed2006

Neural calcitropic peptides: immunoreactive characterization in fish and invertebrates.

Parathyroid hormone (PTH) and stanniocalcin (STC) are primarily produced by the parathyroid glands and corpuscles of Stannius in tetrapods and fish, respectively. However, it is now known that both calcitropic peptides are also synthesized outside of these specialized endocrine glands. The current study employed Western blot analysis to characterize PTH and STC in neural tissues of high- (rats) and low- (hagfish, dogfish, rockfish, trout and skate) vertebrates and invertebrates (starfish, squid, cuttlefish, snails, prawns). Immunoreactive PTH-like peptides, comparable in size to PTH 1-84, were readily detectable in brains of vertebrates lacking (fish) and possessing (rat) parathyroid glands and in invertebrate (snail) ganglia. Immunoreactive STC-like peptides of varying size were similarly detected in brains of vertebrates lacking (rat) and possessing (fish) corpuscles of Stannius and in invertebrate (snail, prawn) ganglia. STC and PTH may thus have evolved as ancestral neuropeptides.

Animals↗

PubMed2005

Comparative study on two freshwater invertebrates for monitoring environmental lead exposure.

Two freshwater invertebrate organisms, Biomphalaria glabrata and Lumbriculus variegatus, were tested as potential experimental animal models to assess Pb exposure using acute laboratory bioassays. Since long, the enzyme delta-aminolevulinic acid dehydratase (ALA-D) has been recognised as a useful biomarker of Pb exposure and effect. Therefore, determinations of ALA-D activity were performed in the whole body soft tissues of pigmented and non-pigmented gastropods B. glabrata and in the oligochaete L. variegatus. The organisms were exposed to varying concentrations of Pb for 48 h. Levels of Pb in the invertebrates were also analysed. Highly significant negative correlations were found between the enzymatic activity and the levels of Pb exposure, and also between the enzymatic activity and the metal incorporated by the invertebrates. No significant differences were found in the enzyme activity nor in the resulting metal accumulation based on gastropod pigmentation. The values of Pb concentration that produce 50% of inhibition on the enzyme activity (EIC50) were 0.023 and 0.029 mg Pb/L for pigmented and non-pigmented B. glabrata, respectively. A much higher value was found for L. variegatus (0.703 mg Pb/L). The non-observed effect concentration (NOEC) on enzyme activity for the oligochaetes was 0.05 mg Pb/L, about twice the EIC50 calculated for the gastropods. These data showed that both pigmented and non-pigmented B. glabata were much more sensitive organisms than the oligochaetes. The differences in enzyme inhibition could be attributed, at least partly, to differences in the metal body burden found between the organisms, since levels of Pb accumulated by B. glabrata were approximately three times higher compared to those observed in L. variegatus. Therefore B. glabrata showed to be a more suitable and reliable bioindicator organism for assessing Pb contamination in aquatic ecosystems, especially at low levels of metal exposure.

Animals↗

PubMed2006

Inhibition of cholinesterase activity by azinphos-methyl in two freshwater invertebrates: Biomphalaria glabrata and Lumbriculus variegatus.

In this study, some biochemical features and the extent of inhibition induced by the organophosphorous pesticide azinphos-methyl on the cholinesterase (ChE) activity present in whole soft tissue of two freshwater invertebrate species, the gastropod Biomphalaria glabrata and the oligochaete Lumbriculus variegatus were investigated. Both invertebrate organisms presented marked differences in ChE activity, type of enzymes and subcellular location. Acetylthiocholine was the substrate preferred by B. glabrata ChE. The enzyme activity was located preferentially in the supernatant of 11,000 x g centrifugation and was inhibited by increasing concentrations of substrate but not by iso-OMPA. Results showed that there were progressive inhibitions of the enzyme activity, with values 21%, 59%, 72%, 76%, and 82% lower than the control at levels of 1, 10, 50, 100 and 1000 microM of eserine, respectively. In contrast, L. variegatus ChE activity was distributed both in the supernatant and pellet fractions, with values approximately 6 and 20 times higher than B. glabrata, respectively. Studies with butyrylthiocholine and iso-OMPA suggested that about 72% of the activity corresponded to butyrylcholinesterase. A strong enzyme inhibition (88-94%) was found at low eserine concentrations (1-10 microM). ChE activity from L. variegatus and B. glabrata was inhibited by in vivo exposure to azinphos-methyl suggesting that both species can form the oxon derivative of this pesticide. However, both invertebrate species showed a very different susceptibility to the insecticide. The NOEC and EIC50 values were 500 and 1000 times lower for L. variegatus than for B. glabrata, reflecting that the oligochaetes were much more sensitive organisms. A different pattern was also observed for the recovery of the enzymatic activity when the organisms were transferred to clean water. The recuperation process was faster for the oligochaetes than for the gastropods. Mortality was not observed in either of the experimental conditions assayed, not even at concentrations that induced 90% of ChE inhibition. The differences in substrate specificity, sensitivity to inhibitors, and subcellular location between the ChEs of B. glabrata and L. variegatus could be the main factors contributing to the differential susceptibility to azinphos-methyl ChE inhibition found in the present study.

Animals↗

PubMed2004

Ecological and evolutionary implications of immunological priming in invertebrates.

Invertebrates have an immune response that differs considerably from the acquired immune response found in vertebrates. However, new studies indicate that past experience with a pathogen can provide individual invertebrates, or their descendants, with enhanced immunity. This prophylactic effect, termed immunological priming, is functionally similar to the acquired immune response in vertebrates. This newfound complexity of invertebrate immunity begs investigation into the conditions under which immunological priming should evolve, and its consequences for population dynamics.

Journal Article↗

PubMed2006

Mating and immunity in invertebrates.

Mating and immunity are intimately linked to fitness. In both vertebrates and invertebrates, recent investigations into mate choice for immunity, tradeoffs between reproduction and immunity, and the relationships between post-mating processes and immune function have revealed that mating and immunity are also intimately linked to each other. Here, we focus on invertebrates and critically examine the evidence that immunity is under sexual selection, both pre- and post-mating, and explore other hypotheses linking mating and immunity. We find little evidence for a consensus regarding which theories best account for the accumulating empirical data. However, we suggest that progress can quickly be made by exploiting the intrinsic strengths of invertebrate model systems.

Animals↗

PubMed2003

A miniature radioimmunoassay for melatonin for use with small samples from invertebrates.

Melatonin has been identified in a number of invertebrates, but little is known of its functions or its circadian control. Studies in this area are hindered by the lack of an assay for melatonin that is appropriate for use with the small sample volumes obtainable from most invertebrates and with the large numbers of sequential samples generated by circadian studies. We describe and validate a novel, miniature radioimmunoassay (RIA) protocol for these purposes. Sample volumes of c.50 microl were extracted with methanol and the extracts dried in 6 x 50 mm glass tubes. The entire assay was conducted in these tubes. The incubation volume was 200 microl. Bound ligand was precipitated and washed in these tubes, then 400 microl scintillation fluid was added and the tubes were mounted inside conventional scintillation vials for counting. Losses of material due to transfer between containers were thus eliminated. Previous RIAs for melatonin employed multiple transfers between containers and their use with small sample volumes can result in loss of substantial proportions of the sample. Two hundred samples could be processed together. The assay sensitivity was 2 pg melatonin and recovery of internal standards was c.100%. Two commercially available antisera gave closely similar results. The assay was used to quantify melatonin titers in the haemolymph of the insect Rhodnius prolixus (Hemiptera); scotophase haemolymph contained 270 pg/ml melatonin, compared with minimal values (c.40 pg/ml) during the photophase, revealing a daily rhythm. This assay opens the door for analyses of the functions and circadian regulation of melatonin in invertebrates.

Animals↗

PubMed2000

Drifting algal mats as an alternative habitat for benthic invertebrates: Species specific responses to a transient resource.

Patchy occurrences of benthic drift algae (i.e. loose lying macroalgal mats) may increase habitat complexity on normally bare soft bottoms, but at the same time, extensive amounts of drifting algal mats are known to stress the benthic fauna. This paper presents results of the first detailed study of the fauna associated with drift algal mats in the northern Baltic Sea. In order to assess the importance of drifting algae as an alternative habitat for benthic fauna, benthic drift algal mats were sampled on shallow (2-9 m) sandy soft bottoms in the outer archipelago of the Åland Islands (Finland). Species composition, abundance and biomass of the macrofauna associated with algal mats were recorded. The results show that drifting algae at times can harbour very high abundances of invertebrates (up to 1116 individuals/g algal dryweight), surpassing invertebrate densities recorded in seagrass communities. The algal fauna varied between sites and over time, and factors such as ambient benthic fauna, exposure to wind-wave disturbance, depth, and algal coverage and condition influenced the invertebrate community composition of the algal mats. Abundance increased while individual biomass of the animals decreased over time (summer season; July-October). A series of laboratory experiments were conducted in order to test the ability of a few important benthic species to move up into, and survive in a drifting algal mat. Macoma balthica, Hydrobia spp., Nereis diversicolor and Bathyporeia pilosa were used in the experiments, and significant differences in their survival and mobility within drifting algae were recorded. This study shows that benthic species differ significantly in their ability to utilise the algal mats, with mainly opportunistic and mobile taxa such as Hydrobia spp., Chironomidae and Ostracoda benefiting from the algae, whereas infaunal species such as M. balthica and B. pilosa are negatively affected. The occurrence of eutrophication induced drifting macroalgal mats has increased significantly during the last decade in the northern Baltic Sea. Hence, the importance of drifting algae as a stress factor and as an alternative habitat for benthic fauna increases.

Journal Article↗

PubMed2001

Transgenic invertebrate models of age-associated neurodegenerative diseases.

Transgenic Drosophila melanogaster and Caenorhabditis elegans strains have been engineered to express human proteins associated with neurodegenerative diseases. These model systems include transgenic animals expressing beta-amyloid peptide (Alzheimer's disease), polyglutamine repeat proteins (Huntington's disease, Spinocerebellar ataxia), and alpha-synuclein (Parkinson's disease). In most of these invertebrate models, some aspects of the human diseases are reproduced. Although expression of all these proteins in transgenic mice has been instructive, the invertebrate models offer experimental advantages (e.g. forward genetic screens) that can potentially address some of the outstanding questions regarding the cellular processes underlying these diseases. This review considers what has been learned from these invertebrate models, and speculates what further insight may be gained from them.

Alzheimer Disease↗

PubMed1990

Homologs of vertebrate growth factors in Drosophila melanogaster and other invertebrates.

The molecular characterization of a number of loci that control developmental processes in invertebrates has revealed that a subset of these genes encode products that are homologous to vertebrate growth factors. Genetic analyses of the autonomy of action and molecular analysis of the patterns of expression of these genes have demonstrated that products of some of these loci (e.g., the EGF homologs, Notch, Delta, lin-12, and glp-1) appear to act in a cell-autonomous manner, while the products of other such loci (e.g., the TGF-beta homolog decapentaplegic and the murine int-1 homolog wingless) act in a nonautonomous manner. Studies of a number of invertebrate EGF homologs, including Notch, Delta, lin-12, and glp-1, for which we are beginning to achieve some reasonable understanding, reveal three common themes. First, each of these loci had been implicated in the determination of cell fates. The products of these loci appear to act at the level of single cells (i.e., they are required for the local choice between alternative determined states). The action of each of these loci within the context of determinative processes is clearly pleiotropic; mutations in each of these genes are correlated with multiple developmental defects. Second, the preponderance of evidence indicates that products of each of these loci function in a cell-autonomous manner during development. This shared character implies that these loci do not encode precursors of EGF-like molecules that act, in turn, as diffusible effectors in determinative decisions. It appears, rather, that these molecules function in association with the membranes of the cells in which they are produced and may constitute components of a class of receptors required for sensing diverse cues that specify particular cell fates during development. Third, we propose that EGF-like sequences found within each of these products function as protein-protein contact motifs that are essential for intermolecular interactions that involve membrane-bound molecules and are central to determinative decisions during development. Assignment of such a function to these sequences is consistent with recent findings indicating that EGF-homologous sequences found in urokinase (Apella et al., 1987) and blood coagulation factor IX (Rees et al., 1988) constitute sites that are required for binding to appropriate interacting proteins and are distinct from the respective "active" sites of each molecule. Within the context of this proposal, products of the EGF-homologous invertebrate genes noted above would participate in the transfer of information required for the specification of cell fate from the extracellular compartment to the cell interior.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acid Sequence↗

PubMedDate not supplied

The venerable inveterate invertebrate TRP channels.

The transient receptor potential (TRP) superfamily is subdivided into four main classes of cation channels, TRPC, TRPV, TRPM and TRPN, each of which includes members in worms, flies, mice and humans. While the biophysical features of many of the mammalian channels have been described, relatively little is known concerning the biological roles of these channels. Forward genetic screens in Drosophila melanogaster and Caenorhabditis elegans have led to the identification of the founding members of each of these four subfamilies. Moreover, phenotypic analyses of invertebrate mutants have contributed greatly to our understanding of the roles of TRP proteins. A recurring theme is that many of these proteins function in sensory signaling processes ranging from vision to olfaction, osmosensation, light touch, social feeding, and temperature- and mechanically-induced nociception. In addition, at least one invertebrate TRP protein is required for cell division. As many of these functions may be conserved among the mammalian TRPs, the invertebrate TRPs offer valuable genetic handles for characterizing the functions of these cation channels in vivo.

Animals↗

PubMed1999

Comparative analysis of invertebrate Tc6 sequences that resemble the vertebrate V(D)J recombination signal sequences (RSS).

Invertebrate cells lack the p53 recombination checkpoint but contain mobile DNA sequences that transpose by a mechanism in part shared with excision of the V(D)J recombination signal sequences (RSS). In this work, inversion, deletion, and duplication of sequences associated with an invertebrate C. elegans Tc6 element is described. The structure of this C. elegans sequence and other dispersed Tc6 elements suggests that covalently closed 'hairpin' structures are not unique to excision of the V(D)J RSS by the RAG proteins, but rather can be generated by transposases at transposon termini leading to characteristic inversion and duplication events. Comparative analysis of recombination events at invertebrate sequences resembling the vertebrate V(D)J RSS may be useful in understanding V(D)J recombination-mediated recombination events in malignant vertebrate cells or genetic diseases such as ataxia telangectasia, in which the p53 recombination checkpoint is defective.

Animals↗

PubMed2000

Cytochrome P450 enzymes in aquatic invertebrates: recent advances and future directions.

A variety of enzymes and other proteins are produced by organisms in response to xenobiotic exposures. Cytochrome P450s (CYP) are one of the major phase I-type classes of detoxification enzymes found in terrestrial and aquatic organisms ranging from bacteria to vertebrates. These enzymes metabolize a wide variety of substrates including endogenous molecules (e.g. fatty acids, eicosenoids, steroids) and xenobiotics (e.g. hydrocarbons, pesticides, drugs). Aquatic invertebrates, especially those in marine habitats, occupy every aspect of the environment, from above the surface (intertidal) to below the sediments. In turn, they have extremely diverse physiologies and are exposed to a vast array of potential toxicants. Aspects of aquatic invertebrate cytochrome P450 enzymes have been studied for the last 25 years. In a few phyla, P450 activities have been measured and are responsive to xenobiotic exposures. Until the last several years, little progress had occurred in the identification of P450 gene diversity in aquatic invertebrates. Molecular biology tools have greatly aided this search, and are likely to identify as much diversity for this protein superfamily as is present in higher marine and terrestrial organisms. Recent work has expanded our knowledge of the CYP superfamily, and new developments will rapidly advance the usefulness of these genes into such fields as biomarker research. Advances of the last decade are reviewed and insights are presented from related insect studies.

Journal Article↗

PubMed2002

Relating cadmium concentrations in three macrophyte-associated freshwater invertebrates to those in macrophytes, water and sediments.

We found that Cd concentrations in three species of macrophyte-associated invertebrates (the gastropods Bithynia tentaculata and Physa gyrina, and the amphipod Gammarus fasciatus) collected at twenty sites along the St. Lawrence River were correlated with Cd concentrations in their main food source, i.e. macrophytes and associated periphyton. Cd in these invertebrates was not significantly correlated with Cd concentrations in the sediments (even when corrections for iron oxide or organic carbon content were applied) or to calculated free-cadmium concentrations in the water. Cd levels in juveniles of B. tentaculata were very tightly linked to Cd concentrations in macrophytes, which is consistent with the close relationship that these organisms have developed with macrophytes. Linear models predicting Cd levels in juveniles of B. tentaculata from Cd levels in different macrophyte species were strong (R2 = 0.69-0.90). Analysis of covariance on these models showed no statistical difference of slope or intercept for any of the macrophyte species, except Vallisneria americana. We suggest that the macrophyte-periphyton complex is a key link in the transfer of cadmium to some aquatic invertebrates in the littoral zone of the St. Lawrence River and that macrophytes and their associated epiphytes should also be used as biomonitors.

Animals↗

PubMed1997

The effects of conservation tillage practices on benthic invertebrate communities in headwater streams in southwestern Ontario, Canada.

Samples of benthic invertebrates from four pairs of streams in southwestern Ontario collected monthly from May through August 1991, and once during the subsequent autumn and winter, demonstrated that conservation tillage practices have a remedial effect on the water quality of adjacent streams as indicated by biotic, rather than physical or chemical, parameters. One stream of each pair drained a basin under conventional tillage (CONV, mainly mouldboard ploughing), the other primarily under conservation tillage (CONS). The paired drainage basins were otherwise similar to one another in type of crops grown, as well as size, topography, soils and hydrology. CONS streams yielded a greater variety of Insecta but fewer taxa of Mollusca, Annelida and Crustacea than did samples from CONV streams. Both kick-net and Surber samples from CONS streams yielded significantly more taxa, more kinds of Ephemeroptera, Plecoptera and Trichoptera, and lower Hilsenhoff Biotic Index scores than did samples from CONV streams. Kick samples from CONS streams were significantly more similar to expected reference communities in overall taxonomic composition. Surber samples from CONS streams contained significantly larger numbers of invertebrates. The relatively greater abundances of infaunal species, especially Tubificidae and Chironomini, in CONV streams suggest greater accumulation of fine sediment particles. Conditions during low flow appear to have the greatest influence on the composition of benthic invertebrate communities. The results of the study indicate that conservation tillage practices have a beneficial effect on the quality of surface waters.

Journal Article↗