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A technique for processing mucous coated marine invertebrate spermatozoa for scanning electron microscopy.

A technique for preparing heavily mucous coated marine invertebrate spermatozoa for scanning electron microscopy (SEM) is described. This technique involves washing in 1500 NF units/ml hyaluronidase in millipored sea water to remove mucus, followed by fixation in glutaraldehyde and osmium tetroxide. Following primary fixation, spermatozoa are enclosed in Nuclepore membrane bags positioned within Teflon specimen capsules allowing them to be processed and critical point dried without excessive mechanical damage or loss.

Animals↗

Rapid screening of invertebrate predators for multiple prey DNA targets.

DNA-based techniques are providing valuable new approaches to tracking predator-prey interactions. The gut contents of invertebrate predators can be analysed using species-specific primers to amplify prey DNA to confirm trophic links. The problem is that each predator needs to be analysed with primers for the tens of potential prey available at a field site, even though the mean number of species detected in each gut may be as few as one or two. Conducting all these PCRs (polymerase chain reactions) is a lengthy process, and effectively precludes the analysis of the hundreds of predators that might be required for a meaningful ecological study. We report a rapid, more sensitive and practical approach. Multiplex PCRs, incorporating fluorescent markers, were found to be effective at amplifying degraded DNA from predators' guts and could amplify mitochondrial DNA fragments from 10+ species simultaneously without 'drop outs'. The combined PCR products were then separated by size on polyacrylamide gels on an ABI377 sequencer. New primers to detect the remains of aphids, earthworms, weevils and molluscs in the guts of carabid predators were developed and characterized. The multiplex-sequencer approach was then applied to field-caught beetles, some of which contained DNA from as many as four different prey at once. The main prey detected in the beetles proved to be earthworms and molluscs, although aphids and weevils were also consumed. The potential of this system for use in food-web research is discussed.

Animals↗

Allergens from mites: implications of cross-reactivity between invertebrate antigens.

Group 1 and 2 allergens are probably the most important mite allergens. Some mite allergens, such as tropomyosin, are widely cross-reactive. The prevalence of sensitization to invertebrates cannot be based solely on skin test or serologic data with the allergen extracts currently available. Purified (recombinant) allergens, selected for low cross-reactivity, are urgently needed.

Allergens↗

Invertebrate neuropeptide hormones.

The development of a long-term research program on the neurosecretory hormones of arthropods is described. The purification and full characterization of the first invertebrate neurohormones, the red pigment-concentrating hormone (RPCH) and the distal retinal pigment hormone (DRPH) demonstrated that they are peptides, an octapeptide and an octadecapeptide, respectively. Physiological function studies with the pure hormones and their synthetic preparations showed that the RPCH acts as a general pigment-concentrating hormone (PCH), and that the DRPH, in addition to its light-adaptive function, also constitutes a general pigment-dispersing hormone (PDH). In the regulation of the color-adaptation of the animals, the two hormones act as antagonists. The chromatophorotropic activities are widely distributed within the arthropod neuroendocrine systems. Purification of the pigment-concentrating activities from the locust corpora cardiaca lead to the isolation and characterization of the first insect neurohormones, the adipokinetic hormones (AKH I and AKH II). These two hormones, AKH I being a decapeptide and AKH II being an octapeptide, are close structural analogs to the crustacean PCH, demonstrating a common evolution of arthropod neurohormones. The hormones of this PCH-family all cross-react, but structure-function studies of the hormones show that quite different parts of their structure are involved in their binding to the various receptors.

Animals↗

Beyond macronutrients: element variability and multielement stoichiometry in freshwater invertebrates.

We contrasted concentrations of macronutrients (C, N and P), essential (As, Cu, Zn and Se) and non-essential metals (Pb, Hg and Cd) in invertebrates across five lakes and June to October in one lake. We predicted that somatic concentrations of tightly regulated elements would be less variable than weakly and unregulated elements. Within each taxon, variation was lowest in macronutrients, intermediate in essential micronutrients, and highest in non-essential metals, which corresponded in rank to homeostatic regulation strength for the same elements calculated from the literature. Hence, homeostatic regulation may strongly influence variation in element concentrations of biota in situ. Of the individual elements, only taxonomic differences in C and N were consistent across lakes and over a season. Nevertheless, canonical discriminant analyses successfully discriminated among taxa based on taxonomic multielement composition. Thus, relative taxonomic differences in multielement composition appear more informative than absolute stoichiometric formulae when considering the role of inherently variable trace elements in ecological investigations.

Animals↗

Vibrio harveyi: a significant pathogen of marine vertebrates and invertebrates.

Vibrio harveyi, which now includes Vibrio carchariae as a junior synonym, is a serious pathogen of marine fish and invertebrates, particularly penaeid shrimp. In fish, the diseases include vasculitis, gastro-enteritis and eye lesions. With shrimp, the pathogen is associated with luminous vibriosis and Bolitas negricans. Yet, the pathogenicity mechanisms are imprecisely understood, with likely mechanisms involving the ability to attach and form biofilms, quorum sensing, various extracellular products including proteases and haemolysins, lipopolysaccharide, and interaction with bacteriophage and bacteriocin-like substances.

Animals↗

Olfactory development in invertebrates. On the scent of central developmental issues.

Invertebrate olfactory systems offer many advantages for cellular and molecular studies of development and for functional studies of developmental plasticity. For example, nematodes have chemical senses that can be studied using genetic approaches. Arthropods, which include insects and crustacea, have the advantages that certain neurons can be reliably identified from one individual to another, and that olfactory receptor neurons are located on peripheral appendages and thus can be manipulated independently of their brain targets even very early in development. Among the insects, olfactory learning can be displayed and used as a basis for studying olfactory plasticity in bees; genes are especially tractable in flies; individual growth cones can be visualized in the grasshopper embryo; and receptor neurons and glomeruli of known olfactory specificity and behavioral significance can be followed during early development in moths. In addition, many insect nervous systems are amenable to organ culture and dissociated-cell culture, opening the door to experimental studies of cellular interactions that can not be performed in situ. Recent research in the moth Manduca sexta attempts to identify the nature of the interactions between olfactory sensory axons, olfactory neurons of the brain, and glial cells in the creation of the array of glomeruli that underlie olfaction in the adult. Results indicate that timing of the ingrowth of olfactory receptor axons is critical for normal glomerulus development, that a subset of axons expresses a fasciclin II-like molecule that may play a role in guidance of their growth, and that glial cells must surround developing glomeruli in order to stabilize the 'protoglomerular' template made by receptor axon terminals. Moreover, glial cells are dye-coupled to each other early in glomerulus development and gradually become uncoupled. Electrical activity in neurons is not necessary for glomerulus formation; and some intercellular interactions, perhaps involving soluble factors, appear to involve tyrosine phosphorylation. In sum, a detailed picture is emerging of the cellular interactions that lead to the formation of glomeruli.

Animals↗

Statistical independence of escalatory ecological trends in Phanerozoic marine invertebrates.

Ecological interactions, such as predation and bioturbation, are thought to be fundamental determinants of macroevolutionary trends. A data set containing global occurrences of Phanerozoic fossils of benthic marine invertebrates shows escalatory trends in the relative frequency of ecological groups, such as carnivores and noncarnivorous infaunal or mobile organisms. Associations between these trends are either statistically insignificant or interpretable as preservational effects. Thus, there is no evidence that escalation drives macroecological trends at global and million-year time scales. We also find that taxonomic richness and occurrence data are cross-correlated, which justifies the traditional use of one as a proxy of the other.

Animals↗

Olfactory reception in invertebrates.

Recent progress in understanding the principles and mechanisms in olfaction is the result of multidisciplinary research efforts that explored chemosensation by using a variety of model organisms. Studies on invertebrates, notably nematodes, insects, and crustaceans, to which diverse experimental approaches can be applied, have greatly helped elucidate various aspects of olfactory signaling. From the converging results of genetic, molecular, and physiological studies, a common set of chemosensory mechanisms emerges. Recognition and discrimination of odorants as well as chemo-electrical transduction and processing of olfactory signals appear to be mediated by fundamentally similar mechanisms in phylogenetically diverse animals. The common challenge of organisms to decipher the world of odors was apparently met by a phylogenetically conserved strategy. Thus, comparative studies should continue to provide important contributions toward an understanding of the sense of smell.

Action Potentials↗

Protein translocation in photoreceptor light adaptation: a common theme in vertebrate and invertebrate vision.

How do our eyes adjust to daily light levels that vary by almost 11 orders of magnitude? Research shows that, in both vertebrates and invertebrates, signaling proteins are translocated in a light-dependent manner between the photoreceptor cellular compartments where visual transduction takes place, and the rest of the photoreceptor cell. Protein translocation is likely to contribute to photoreceptor light adaptation by adjusting the sensitivity and speed of photoresponse to ever-changing conditions of ambient illumination.

Adaptation, Ocular↗

Crustacean-insect relationships: the use of brain characters to derive phylogeny amongst segmented invertebrates.

Conserved neural characters identified in the brains of a variety of segmented invertebrates and outgroups have been used to reconstruct phylogenetic relationships. The analysis suggests that insects and crustaceans are sister groups and that the 'myriapods' are an artificial construct comprising unrelated chilopods and diplopods. Certain elements of the optic lobes and mid-brain support the notion that insects are more closely related to crustaceans than they are to any other arthropods. However, deep optic neuropils and optic chiasmata are homoplastic in insects and crustaceans. The organization of olfactory pathways suggests that insect olfactory lobes originated late, probably first appearing in orthopteroid or blattoid pterygotes. The present results are discussed with respect to recent studies on early development of arthropod nervous systems and the fossil record.

Animals↗

Oncoprecipitins from marine invertebrates are glycoproteins with a higher specificity to carcinoembryonic antigen.

Oncoprecipitins isolated from a number of marine invertebrates have been shown to be glycoproteins possessing a higher specificity to carcinoembryonic antigen (CEA)--similar to the interaction of antigen-antibody. The oncoprecipitin-CEA interaction was found to be substantially more specific than that of carbohydrate-containing polymers with lectins. Two oncoprecipitins, crustacin and cyprein, isolated from Pagurus prideauxii and Cyprea caputserpentis, respectively, have been characterized. The binding reaction of CEA with crustacin and cyprein failed to be inhibited with glycoproteins having sugar chain patterns closely related to those of CEA. Inhibition of the oncoprecipitin-CEA interaction with sugars was shown to be nonspecific and was observed only at sufficiently high concentrations of sugars used as inhibitors. Immunoenzymic assay of CEA content in normal and tumor tissues using CEA crustacin and CEA antiserum against CEA test systems revealed highly consistent results for both systems.

Animals↗

Invertebrate paraxial locomotory appendages: design, deformation and control.

Some principles governing the design of invertebrate paired propulsive appendages are discussed, with particular reference to the extent to which information encoded in their skeletal structure determines their instantaneous shape in locomotion. The hydrostatic paired fins of some cephalopods and marine gastropods, polychaete parapodia and onychophoran lobopodia rely entirely on musculature for shape control. The deformations of walking limbs, though still under muscular control, are strongly influenced by the nature and sequence of movement of the joints. Limbs adapted for walking in air are effectively point-loaded, and their rigid components need to resist axial forces as well as bending and torsional moments. Aquatic walking limbs have little axial loading, while swimming appendages and wings experience only bending and torsional moments, and can exploit these to assist in the deformations that are necessary to gain force asymmetry between half-strokes. Swimming appendages normally employ both muscles and drag, but the wings of insects lack internal muscles, and their changes in shape are largely complex aeroelastic responses to the constantly changing aerodynamic and inertial loads, moderated by muscles inserted at the base. For illustration, wings modelled as thin shells with flexible hinge-lines are used to investigate how transverse distal flexion, essential for controlling the angle of attack in the upstroke, is remotely controlled by the indirect flight muscles.

Animals↗

The colloid osmotic pressures of invertebrate body fluids.

Colloid osmotic pressures of the body fluids of twenty invertebrate species were measured directly. The results, which are generally lower than predicted values for the same species, pertain to several physiological questions: (1) they do not quantitatively explain the frequently observed hyperosmoticity of body fluids in species believed to be osmoconformers, indicating that the condition cannot be merely a consequence of a Gibbs-Donnan equilibrium; (2) the excess of hydrostatic over colloid osmotic pressure is very small. This result supports the hypothesis that the oxygen transport function of bloods with extracellular haemocyanins and haem proteins is limited by their colligative properties; (3) the pressure relationships and the absence of colloid osmotic activity in urine indicates that filtration contributes to urine formation in several species.

Animals↗

The parkinsonian models: invertebrates to mammals.

In contrast to Alzheimer's disease, effective therapeutic options are available for Parkinson's disease. Therapy of dopamine replacement such as levodopa improves the symptoms of this disease, but does not inhibit neurodegeneration in the substantia nigra. Numerous studies have suggested that endogenous and environmental neurotoxins, and oxidative stress may participate in this disease, but the detailed mechanisms are still unclear. Recent genetic studies in familial Parkinson's disease and parkinsonism show several gene mutations. This new information regarding pathogenesis offers novel prospects for therapy. To develop novel neuroprotective drugs, it is necessary to have a model for each type of parkinsonism. This review summarizes current findings regarding parkinsonian models in vertebrates and invertebrates and discusses their value.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

Absence of endothelium in invertebrate blood vessels: significance of endothelium and sympathetic nerve/medial smooth muscle in the vertebrate vascular system.

In the course of evolution, two remarkable changes seem to have occurred in vertebrate circulation: the appearance and development of the "endothelium or endothelial tubular system" and "sympathetic nerve/medial smooth muscle system". In the present article, some relevant literature is reviewed and discussed. Absence of endothelium in the vascular wall of most invertebrates had been known and was confirmed by recent electron microscopic studies. The medial smooth muscle is rather proper to vertebrate vessels. It seems to have appeared after emergence of and in association with the endothelial tubular system. Phylogenetically, the parasympathetic nervous system is thought to be older than the sympathetic system. The former is distributed to viscera and the latter developed in close relation with the vascular system. It is assumed that during evolution, a circulatory system composed of the heart and endothelial tubular system first formed in vertebrates, medial smooth muscle then appeared for regulation of the system, and innervation of the muscle tissue took place. This sequence of development assumed for phylogenesis is actually realized in the ontogenetic processes. We thus propose a hypothesis that the "sympathetic nerve/medial smooth muscle system" may be regarded as a new neuroeffector mechanism that developed for systemic regulation of the endothelium-lined closed vascular system in vertebrates. A few implications of the hypothesis are presented.

Animals↗

The activator of human cerebroside sulphatase. Activating effect on the acidic forms of the sulphatases from invertebrates.

1) Acidic forms of the sulphatase were partially purified from the following invertebrate species: Tethya aurantium (Porifera), Patella vulgata (mollusca), Maja squinado (Arthropoda), Marthasterias glacialis (Echinodermata) and Microcosmus sulcatus (Tunicata). Enzyme preparations thus obtained cleaved cerebroside sulphates (sulphatides) only in the presence of either specific detergents (e.g. taurodeoxycholate) or an activator protein isolated from human liver. This corresponds to the findings on purified sulphatase A of human origin. 2) At low concentrations, the activating effect was proportional to the amount of activator protein applied; at higher concentrations, proportionality was obtained only in some cases. On a molar basis, less of the activator protein was required to achieve the same activation as taurodeoxycholate. At optimum concentrations of the detergent however, the activation was much higher. 3) The enzyme specificity of the activator and some evolutionary implications are discussed.

Acetylglucosaminidase↗

An invertebrate histocompatibility complex.

We have developed defined genetic lines of the hydroid Hydractinia symbiolongicarpus and confirmed earlier results showing that allorecognition is controlled by a single chromosomal region within these lines. In a large backcross population, we detected recombinants that display a fusibility phenotype distinct from typical fusion and rejection. We show that this transitory fusion phenotype segregates in a fashion expected of a single Mendelian trait, establishing that the chromosomal interval contains at least two genes that interact to determine fusibility. Using bulked segregant analysis, we have identified amplified fragment length polymorphisms (AFLP) cosegregating with fusibility, used these markers to independently confirm linkage of the two loci, and constructed a 3.4-cM map of an invertebrate histocompatibility complex.

Alleles↗