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[A new family of Nematodes, parasites of marine invertebrates, the Benthiumermithidae (author's transl)].

Nematodes of a group related to the Mermithoidea and known only from a preliminary note by Hope (1977) have been found either as parasites of invertebrates (only female juveniles) or free-living (mature and juvenile females) in benthic sediments of the Atlantic Ocean, the Norway Sea, and the Indian Ocean. The characters in common in the group are given. Benthimermis hopei, n. g., n. sp., known from one mature and one juvenile females found free in benthic sediments of the Atlantic Ocean is described, as well as 8 juvenile females found free in benthic sediments of the Atlantic Ocean is described, as well as 8 juvenile females (Benthimermis sp., nos 1-8), of which 7 are parasites of invertebrates and one was found free in sediments. A new family is proposed for the group, the Benthimermithidae, characterized by a vestigial oesophagus, reduced to a strand containing granular material, an intestine modified into a trophosome, and a vestigial rectum and anus.

Animals↗

Heterochromatin (chromocentres) and regeneration of invertebrates and vertebrates: a correlation.

Previous investigations using several species of invertebrates have indicated a correlation between nuclear chromocentre content and the ability to regenerate. Such findings have resulted in the formulation of the hypothesis that the ability to regenerate may be dependent on cellular heterochromatin content. In order to test this concept further, a number of additional invertebrates were stained for heterochromatin (nuclear chromocentres). In accordance with the hypothesis, each species tested was found to contain numerous chromocentres. Many chromocentres were also found in cells of several vertebrates capable of partial regeneration, thereby extending the hypothesis to vertebrate regeneration. These findings provide additional supporting evidence for a correlation between cellular heterochromatin and the ability to regenerate.

Amphibians↗

Antiport-driven sulfate secretion in an invertebrate epithelium.

A novel invertebrate gastrointestinal transport mechanism has been shown to couple chloride/sulfate exchange in an electrogenic fashion. In the lobster, Homarus americanus, the hepatopancreas, or digestive gland, exists as an outpocketing of the digestive tract, representing a single cell layer separating the gut lumen and an open circulatory system comprised of hemolymph. Investigations utilizing independently prepared brush-border and basolateral membrane vesicles revealed discrete antiport systems which possess the capacity to bring about a transcellular secretion of sulfate. The luminal antiport system functions as a high affinity, one-to-one chloride-sulfate exchanger that is stimulated by an increase in luminal hydrogen ion concentration. Such a system would take advantage of the high chloride concentration of ingested seawater, as well as the high proton concentrations generated during digestion, which further suggests a potential regulation by resident sodium-proton exchangers. Exchange of one chloride for one divalent sulfate ion provides the driving force for electrogenic vectorial translocation. The basolateral antiport system was found to be electroneutral in nature, responsive to gradients of the dicarboxylic anion oxalate, while lacking in proton stimulation. No evidence of sodium-sulfate cotransport, commonly reported for the brush border of vertebrate renal and intestinal epithelia, was observed in either membrane preparation. The two antiporters together can account for the low hemolymph to seawater sulfate levels previously described in decapod crustaceans. A secretory pathway for sulfate based upon electrogenic chloride-antiport may appear among invertebrates partly in response to digestion taking place in a seawater environment.

Animals↗

Sequence of a functional invertebrate GABAA receptor subunit which can form a chimeric receptor with a vertebrate alpha subunit.

The sequence of an invertebrate GABAA receptor subunit is described. This was deduced from a cDNA which was isolated from the mollusc Lymnaea stagnalis and which corresponds to a transcript of extremely low abundance. The cDNA was isolated using short exonic sequences from part of the corresponding gene in combination with a variant of the polymerase chain reaction (PCR) known as RACE (rapid amplification of cDNA ends). The mature polypeptide has a predicted molecular weight of 54,569 Daltons and exhibits approximately 50% identity to vertebrate GABAA receptor beta subunits. The six intron-exon boundaries determined to date in the molluscan gene occur at the same relative positions as those found in vertebrate GABAA receptor genes. Functional expression, in Xenopus oocytes, of the molluscan cDNA alone results in the formation of GABA-activated chloride ion channels that have a finite open probability even in the absence of agonist. These GABA-evoked currents can be reversibly blocked by the vertebrate GABAA receptor antagonist bicuculline. Surprisingly, the molluscan beta subunit is capable of replacing vertebrate beta subunits in co-expression experiments with the bovine GABAA receptor alpha 1 subunit. These findings suggest that invertebrate GABAA receptors exist in vivo as hetero-oligomeric complexes.

Amino Acid Sequence↗

A common ancestor DNA motif for invertebrate and vertebrate hormone response elements.

The ecdysone-responsive DNA sequence of the Drosophila hsp27 gene promoter contains four direct and inverted repeats reminiscent of those that compose the vertebrate palindromic estrogen response element (ERE) and the thyroid hormone/retinoic acid response element (TRE/RRE). Interestingly, a 3 bp substitution in the wild-type Hsp27 ecdysone response element (EcdRE) increases both its similarity with the vertebrate ERE and TRE/RRE and its capacity to confer ecdysone responsiveness to a heterologous promoter. Remarkably, increasing the spacing between the inverted repeats of this strong EcdRE by two nucleotides converts it into an ERE. Inversely, decreasing the spacing between the two inverted repeats of the vertebrate consensus palindromic ERE, from three to one nucleotide, converts it into a functional EcdRE. Thus, the only difference between an invertebrate EcdRE and a vertebrate palindromic ERE or TRE/RRE is in the spacing between the conserved inverted repeated motifs forming these palindromic HREs. The finding that the sequence motif 5'-GGTCA-3' present in the vertebrate ERE and TRE/RRE is also a functionally important characteristic of an invertebrate HRE, suggests that a common ancestor regulatory DNA sequence gave rise to all HREs known so far. We discuss the possibility that this progenitor motif is the GGTCA sequence.

Animals↗

Phagocytosis by invertebrate hemocytes: causes of individual variation in Panorpa vulgaris scorpionflies.

An in vitro phagocytosis assay, adjusted to as little as 1 microL of insect hemolymph, enables the microscopic determination of phagocytosis for single individuals of small insects. Even repeated determination over the lifetime of individuals is possible. This method makes it feasible to study individual variation in invertebrate phagocytic capacity. Possible sources of such variation are reviewed in this article: genetic differences, development, aging, reproduction, presence of parasites, and diverse environmental influences are natural sources of individual variation in phagocytosis. However, the methods used for phagocytosis and microscopic evaluation are also (unwelcome) sources of variation. To optimize incubation time for in vitro phagocytosis, time courses were determined. Furthermore, the reliability of visual counting and image analysis for the microscopic quantification of phagocytosis are compared. The influences of larval development and adult aging on phagocytosis by Panorpa vulgaris hemocytes are subsequently demonstrated. During development, a decrease in hemocyte numbers but a simultaneous increase in the proportion of phagocytosing hemocytes was observed when larvae reached pupation. On the other hand, adults showed a dramatic decrease in phagocytic capacity with age, while cell numbers remained fairly constant. The results show that individual variation in phagocytosis can be determined accurately in small invertebrates and related to its causes. This might be especially interesting in the context of studies relating individual immunocompetence to ecology, life history variation, and behavior.

Aging↗

Contraction increases the T(2) of muscle in fresh water but not in marine invertebrates.

Previous studies suggest that the activity-induced increase in (1)H-NMR transverse relaxation time (T(2)) observed in mammalian skeletal muscles is related to an osmotic effect of intracellular metabolite accumulation. This hypothesis was tested by comparing T(2) (measured by (1)H-NMR imaging at 4.7 T) and metabolite changes (measured by (31)P-NMR spectroscopy) after stimulation in the muscles of a freshwater (crayfish, Orconectes virilis) vs two osmoconforming marine invertebrates (lobster, Homarus americanus; scallop, Argopecten concentricus). Intracellular pH significantly decreased after stimulation in the lobster tail muscle, but not in the crayfish tail or scallop phasic adductor muscles. The decrease in phosphoarginine-to-ATP ratio after stimulation was similar in the three muscles. Muscle T(2) increased from 37 to 43 ms (p < 0.02, n = 7) after stimulation in crayfish, but was unchanged in lobster muscle (32 ms, n = 7), and significantly decreased (from 40 to 36 ms, p < 0.02, n = 11) in scallop muscle. The observation that T(2) does not increase after stimulation in muscles of marine invertebrates with high natural osmolarity is consistent with the hypothesis that the T(2) increase in mammalian muscle is related to osmotically driven shifts of fluid between subcellular compartments.

Adenosine Triphosphate↗

Invertebrate models of drug abuse.

Susceptibility to drug addiction depends on genetic and environmental factors and their complex interactions. Studies with mammalian models have identified molecular targets, neurochemical systems, and brain regions that mediate some of the addictive properties of abused drugs. Yet, our understanding of how the primary effects of drugs lead to addiction remains incomplete. Recently, researchers have turned to the invertebrate model systems Drosophila melanogaster and Caenorhabditis elegans to dissect the mechanisms by which abused drugs modulate behavior. Due to their sophisticated genetics, relatively simple anatomy, and their remarkable molecular similarity to mammals, these invertebrate models should provide useful insights into the mechanisms of drug action. Here we review recent behavioral and genetic studies in flies and worms on the effects of ethanol, cocaine, and nicotine, three of the most widely abused drugs in the world.

Alcoholic Intoxication↗

Invertebrate nociception: behaviors, neurons and molecules.

Genetic analysis of nociceptive behaviors in the nematode Caenorhabditis elegans and the fruit fly Drosophila melanogaster has led to the discovery of conserved sensory transduction channels and signaling molecules. These are embedded in neurons and circuits that generate responses to noxious signals. This article reviews the neurons and molecular mechanisms that underlie invertebrate nociception. We begin with the neurobiology of invertebrate nociception, and then focus on molecules with conserved functions in vertebrate nociception and sensory biology.

Animals↗

Characterization of carbonic anhydrases from Riftia pachyptila, a symbiotic invertebrate from deep-sea hydrothermal vents.

The symbiotic hydrothermal vent tubeworm Riftia pachyptila needs to supply its internal bacterial symbionts with carbon dioxide, their inorganic carbon source. Our aim in this study was to characterize the carbonic anhydrase (CA) involved in CO(2) transport and conversion at various steps in the plume and the symbiotic tissue, the trophosome. A complete 1209 kb cDNA has been sequenced from the trophosome and identified as a putative alpha-CA based on BLAST analysis and the similarities of total deduced amino-acid sequence with those from the GenBank database. In the plume, the putative CA sequence obtained from cDNA library screening was 90% identical to the trophosome CA, except in the first 77 nucleotides downstream from the initiation site identified on trophosome CA. A phylogenetic analysis showed that the annelidan Riftia CA (CARp) emerges clustered with invertebrate CAs, the arthropodan Drosophila CA and the cnidarian Anthopleura CA. This invertebrate cluster appeared as a sister group of the cluster comprising mitochondrial and cytosolic isoforms in vertebrates: CAV, CAI II and III, and CAVII. However, amino acid sequence alignment showed that Riftia CA was closer to cytosolic CA than to mitochondrial CA. Combined biochemical approaches revealed two cytosolic CAs with different molecular weights and pI's in the plume and the trophosome, and the occurrence of a membrane-bound CA isoform in addition to the cytosolic one in the trophosome. The physiologic roles of cytosolic CA in both tissues and supplementary membrane-bound CA isoform in the trophosome in the optimization of CO(2) transport and conversion are discussed.

Amino Acid Sequence↗

Identification of a new member of transforming growth factor-beta superfamily in Drosophila: the first invertebrate activin gene.

Activins, a subgroup of the transforming growth factor-beta (TGF-beta) superfamily, have been extensively studied in vertebrates for their roles in growth and development. However, activins are not thought to be expressed in invertebrates. The identification of the first invertebrate activin gene is reported here. A genomic clone representing 102 F region of the Drosophila chromosome 4 is found to encode a putative activin beta. The predicted protein sequence has a multibasic protease site that would generate a mature C-terminal peptide containing 113 amino acids showing > 60% similarity to the vertebrate activin beta B (inhibin beta B) sequences. A TGF-beta family signature as well as all 9 cysteine residues conserved in the vertebrate activins are also present in this mature peptide sequence. Northern blot and RT-PCR analyses indicated that the activin beta gene is expressed in embryo, larva and adult stages of Drosophila.

Amino Acid Sequence↗

Characterization of a novel cDNA sequence encoding invertebrate tachykinin-related peptides isolated from the echiuroid worm, Urechis unicinctus.

Tachykinin is one of the most well-known bioactive peptides found in vertebrates, and tachykinin-related peptides have also been isolated from various invertebrate species. Urechistachykinin I (Leu-Arg-Gln-Ser-Gln-Phe-Val-Gly-Ser-Arg-NH(2)) and II (Ala-Ala-Gly-Met-Gly-Phe-Phe-Gly-Ala-Arg-NH(2)) were purified from the ventral nerve cords of echiuroid worm, Urechis unicinctus. In the present study, we described the characterization of a novel cDNA encoding the urechistachykinin precursor. Amino acid sequence analysis of the deduced polypeptide revealed that the urechistachykinin precursor included seven structurally related peptides, unlike mammalian tachykinin precursors which encode only one or two tachykinin peptides. This is the first identification of an invertebrate tachykinin-related peptide cDNA.

Amino Acid Sequence↗

Androgens regulate the mammalian homologues of invertebrate sex determination genes tra-2 and fox-1.

Androgens, like other steroid hormones, exert profound effects on cell growth and survival by modulating the expression of target genes. In vertebrates, androgens play a critical role downstream of the testis determination pathway, influencing the expression of sexually dimorphic traits. Among cells of the nervous system, motor neurons respond to trophic effects of androgen stimulation, with a subpopulation of spinal motor neurons exhibiting sexually dimorphic survival. To study the mechanisms of androgen action in these cells, we performed a subtractive screen for genes upregulated by androgen in a motor neuron cell line. We show androgen-inducible expression of two RNA-binding proteins that are the mammalian homologues of invertebrate sex determination genes. Androgens upregulate the expression of tra-2alpha, an enhancer of RNA splicing homologous to Drosophila tra-2, and promote redistribution of the protein from a diffuse to a speckled pattern within the nucleus. Similarly, androgens upregulate the expression of a novel gene homologous to Caenorhabditis elegans fox-1. These data indicate that androgens exert their effects, in part, by modulating the expression and function of genes involved in RNA processing, and identify homologues of invertebrate sex determination genes as androgen-responsive genes in mammals.

Amino Acid Sequence↗

Safety testing of tebufenozide, a new molt-inducing insecticide, for effects on nontarget forest soil invertebrates.

Tebufenozide, a new molt-inducing insecticide that mimics the action of ecdysone, is being considered for use to control defoliating lepidoptera in forests in Canada. Soil microcosms, employing substrates and species from the ecosystems in which spraying is likely to occur, were used to evaluate the effects of this compound on soil invertebrates. The forest earthworm (Dendrobaena octaedra Savigny) and four species of Collembola (Folsomia candida Willem, F. nivalis (Packard), Onychiurus parvicornis Mills, and Hypogastrura pannosa Macnamara) were tested. Survival, growth, and reproduction (cocoon production and viability) in the forest earthworm D. octaedra were not affected by exposure to tebufenozide at concentrations up to and including 100x expected environmental concentration (EEC; equivalent to the operational spray rate; 70 g/ha) in leaf litter over a 10-week period. Similarly, population growth over 8-10 weeks in the four species of soil Collembola in LFH material contaminated with tebufenozide at 100x EEC was not affected. Based on the results of these (limited) tests, it therefore appears that, under normal operational conditions, tebufenozide should not pose a hazard to soil invertebrates. The selection of appropriate test species and systems for forestry applications is discussed.

Aerosols↗

Invertebrate host-parasite relationships: convergent evolution of a tropomyosin epitope between Schistosoma sp., Fasciola hepatica, and certain pulmonate snails.

Monoclonal antibodies (mAb) directed against Schistosoma mansoni tropomyosin isoform, SMTM (Xu et al. Experimental Parasitology 69, 373-392, 1989), were used to test for cross-reactivity with Biomphalaria glabrata antigens. One mAb (1F10) recognized antigens of 39, 41, and 80 kDa in a snail head/foot antigen preparation but not a hepatopancreas antigen preparation. Another mAb (1C1) cross-reacted with a 39-kDa antigen in the head/foot extract but not in the hepatopancreas extract. Epitope mapping revealed the 1F10 epitope to be between amino acids 135 and 188 of both Bg39 (Dissous et al. Molecular and Biochemical Parasitology 43, 245-256, 1990) and BgTMII (Weston and Kemp, Experimental Parasitology 76, 358-370, 1993), while the 1C1 epitope was located between amino acids 189 and 213 of BgTMII. Various invertebrate species, including members from Trematoda, Pulmonata, Annelida, and Arthropoda, were tested for cross-reactivity with the monoclonal antibodies. While the 1F10 mAb displayed broad invertebrate cross-reactivity, the 1C1 mAb cross-reactivity was restricted to schistosomes, F. hepatica, and the pulmonate snails B. glabrata and Physa sp.

Amino Acid Sequence↗

Atrial natriuretic peptide gene expression within invertebrate hearts.

The present investigation was designed to (1) determine if atrial natriuretic factor gene expression occurs within invertebrates as well as within vertebrates; (2) determine whether the product of this gene expression is the 126-amino-acid atrial natriuretic factor prohormone or some other molecular species; and (3) evaluate within the same invertebrates if the products of atrial natriuretic factor gene expression are released into their circulation. Utilizing a very sensitive RNase protection assay it was found that atrial natriuretic peptide gene expression occurs within the heart of the oyster, Crassostrea virginica, and within the heart of the blue crab, Callinectes sapidus, but was expressed sevenfold less than in a vertebrate heart (i.e., rat, Rattus norvegiucs). High-performance gel-permeation chromatography followed by N-terminal and C-terminal atrial natriuretic factor prohormone radioimmunoassays indicated that the molecular species synthesized within the oyster and blue crab hearts was the atrial natriuretic factor prohormone. The product(s) of this atrial natriuretic factor gene expression (i.e., atrial natriuretic peptides) was found to be released into the circulation, i.e., hemolymph, of both the oyster and the blue crab.

Animals↗

Chemical defense in the egg masses of benthic invertebrates: an assessment of antibacterial activity in 39 mollusks and 4 polychaetes.

Many marine invertebrates deposit benthic egg masses that are potentially vulnerable to microbial infection. To help counter this threat these species may have evolved some form of chemical protection for their encapsulated embryos. In this study the egg masses from 7 marine mollusks were tested for antibacterial activity against 4 marine pathogens: Enterococcus sericolicida, Vibrio anguillarum, Vibrio alginolyticus, and Vibrio harveyi. Extracts from all of these egg masses were found to inhibit the growth of at least 1 marine bacterium at concentrations that approximate the natural concentration of extract in the egg masses. The egg masses of 39 mollusks and 4 polychaetes were then tested for antibacterial activity against 3 human pathogenic bacteria; Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. Activity was detected in the egg masses from 34 species, including 2 polychaetes and mollusks from two classes and 18 families. Antibacterial activity in molluskan egg masses was found to extend across the marine, estuarine, freshwater, and terrestrial environments. Both gelatinous egg masses and tough egg capsules were found to inhibit microbial growth, suggesting that physical protection alone may not be sufficient to protect the eggs. Antimicrobial activity was observed in the fresh egg masses but not in the well-developed egg masses of a subset of species. The results of this study indicate that a wide range of invertebrates use chemical defense to protect their early stage embryos against bacterial infection.

Animals↗

Bioactive natural products from marine invertebrates and associated fungi.

Marine natural products with their unique structural features and pronounced biological activities continue to provide lead structures in the search for new drugs from nature. Invertebrates such as sponges, tunicates, mollusks and others that are either sessile or slow moving and mostly lack morphological defense structures have so far provided the largest number of marine-derived secondary constituents including some of the most interesting drug candidates. This review highlights recent research findings of our group related to natural products from marine invertebrates. Areas that are covered include ecological functions of secondary constituents from sponges against predatory fish, the search for new pharmacologically active constituents from sponges and tunicates, and sponge-associated fungi as an evolving source for new bioactive natural products.

Alkaloids↗