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The vertebrate linker histones H1 zero, H5, and H1M are descendants of invertebrate "orphon" histone H1 genes.

We investigated the evolutionary history of the divergent vertebrate linker histones H1 zero, H5, and H1M. We observed that the sequence of the central conserved domain of these vertebrate proteins shares characteristic features with histone H1 proteins of plants and invertebrate animals which otherwise never appear in any vertebrate histone H1 protein. A quantitative analysis of 58 linker histone sequences also reveals that these proteins are more similar to invertebrate and plant histone H1 than to histone H1 of vertebrates. A phylogenetic tree deduced from an alignment of the central domain of all known linker histones places H1 zero, H5, and H1M in close vicinity to invertebrate sperm histone H1 proteins and to invertebrate histone H1 proteins encoded by polyadenylated mRNAs. We therefore conclude that the ancestors of the vertebrate linker histones H1 zero, H5, and H1M diverged from the main group of histone H1 proteins before the vertebrate type of histone H1 was established in evolution. We discuss this observation in the general context of linker histone evolution.

Amino Acid Sequence↗

Invertebrate organisms as biological indicators of heavy metal pollution.

Some species of invertebrate animals are known to be efficient accumulators of trace elements. Generally, metal accumulation by such organisms is based on efficient detoxification mechanisms, such as intracellular compartmentalization, or metal inactivation by binding to metallothioneins. Metal accumulators have often been used as accumulation indicators of environmental metal pollution. This means that, ideally, metal concentrations in the animal's body reflect quantitatively or semiquantitatively environmental pollution levels. In reality, however, many factors, such as the animal's weight and age, can disturb such quantitative relationships. These factors have, therefore, to be considered carefully before an invertebrate is utilized as accumulation indicator for metal pollution. Apart from accumulation, many invertebrates exposed to elevated metal concentrations respond to this stress by metal-induced synthesis of metallothioneins. Additionally, metallothionein in metal-loaded organisms can be present in different isoforms that are specifically synthesized in response to different metals. These facts make metallothionein a potential biomarker for metal stress in invertebrates. One possibility may be to assess parameters of metallothionein synthesis at the molecular or biochemical level. Moreover, metallothionein isoform patterns could provide information on different isoforms synthesized in response to different metals or chemicals. In any case, however, care must be taken to consider intrinsic physiological parameters, such as nutritional or developmental factors, which could also interfere with metallothionein synthesis.

Animals↗

Common evolutionary origin and birth-and-death process in the replication-independent histone H1 isoforms from vertebrate and invertebrate genomes.

The H1 histone multigene family shows the greatest diversity of isoforms among the five histone gene families, including replication-dependent (RD) and replication-independent (RI) genes, according to their expression patterns along the cell cycle and their genomic organization. Although the molecular characterization of the RI isoforms has been well documented in vertebrates, similar information is lacking in invertebrates. In this work we provide evidence for a polyadenylation signature in the Mytilus "orphon" H1 genes similar to the polyadenylation characteristic of RI H1 genes. These mussel genes, together with the sea urchin H1delta genes, are part of a lineage of invertebrate "orphon" H1 genes that share several control elements with vertebrate RI H1 genes. These control elements include the UCE element, H1-box and H4-box. We provide evidence for a functional evolution of vertebrate and invertebrate RI H1 genes, which exhibit a clustering pattern by type instead of by species, with a marked difference from the somatic variants. In addition, these genes display an extensive silent divergence at the nucleotide level which is always significantly larger than the nonsilent. It thus appears that RI and RD H1 isoforms display similar long-term evolutionary patterns, best described by the birth-and-death model of evolution. Notably, this observation is in contrast with the theoretical belief that clustered RD H1 genes evolve in a concerted manner. The split of the RI group from the main RD group must therefore have occurred before the divergence between vertebrates and invertebrates about 815 million years ago. This was the result of the transposition of H1 genes to solitary locations in the genome.

Animals↗

The unusual cartilaginous tissues of jawless craniates, cephalochordates and invertebrates.

A collagenous extracellular matrix was previously considered to be a requirement for classification of true cartilage. Data from the lamprey and hagfish now clearly indicate that both of these jawless craniates have extensive non-collagenous, yet cartilaginous endoskeletons. Non-collagenous cartilages are present in the cephalochordates (amphioxus) and in the invertebrates, although collagen-containing cartilages also are found in the invertebrates. This review summarizes current knowledge of the morphological, biochemical and molecular characteristics of the unusual non-collagenous cartilages in jawless craniates and the cartilaginous tissues in amphioxus and invertebrates. A least two types of non-collagenous cartilage matrix proteins are found in both the hagfishes and the lampreys, all of which are resistant to digestion by cyanogen bromide (CNBr). Although all four of these matrices show some similarities with each other, suggesting a family of non-collagenous, elastin-like proteins, it is clear that the major matrix proteins of each are different. New morphological and biochemical information on the cartilaginous tissues in squid, horseshoe crab and amphioxus reveals the presence of CNBr-insoluble, non-collagenous matrix proteins, potentially extending the jawless craniate family of cartilaginous proteins into the invertebrates. Details of the evolutionary relationships between these non-collagenous matrix proteins and the significance of the occurrence of these proteins as the major components of the cartilaginous tissues of jawless craniates, amphioxus, horseshoe crab and squid, all of which are capable of producing a variety of collagens in other tissues, remain to be investigated.

Amino Acids↗

Sex steroid receptor evolution and signalling in aquatic invertebrates.

In vertebrate reproductive endocrinology sex steroids play a pivotal role via binding to receptors. However, information on the origin and relevance of sex steroids in invertebrates is limited. This review highlights current literature on steroid receptors in aquatic invertebrates and reports on some new findings. It has been shown that invertebrates of the deuterostome clade, such as Acrania and Echinodermata, respond to estrogens and androgens and, at least in Branchiostoma, an estrogen receptor has been cloned. Within the protostomes, most findings are related to aquatic molluscs. Sex steroid receptor-like proteins are abundant in gastropods, bivalves and cephalopods and also sex hormone signalling shows partial similarity to the deuterostomes. In ecdysozoans, however, the impact of sex steroids is still a matter of debate even though there is evidence on the presence of estrogen receptor-like proteins in Crustacea and on physiological effects of estrogens in both Nematoda and Crustacea. Recent findings suggest the presence of an estrogen receptor alpha-like protein of unclear physiological role in Gammarus fossarum (Crustacea). Binding studies revealed the crustacean Hyalella azteca to possess specific binding sites only for androgens but not for estrogens suggesting a possible limitation to functional androgen receptors in this species. Further studies have to be conducted to shed more light into the discussion about the controversy about sex steroid receptors in invertebrates.

Amino Acid Sequence↗

Partial sequence of connectin-like 1200K-protein in obliquely striated muscle of a polychaete (Annelida): evidence for structural diversity from vertebrate and invertebrate connectins.

Vertebrate striated muscle contains the giant elastic protein connectin that maintains the position of the A-band at the center of the sarcomere during repeated muscular contraction and relaxation. Connectin-like molecules may perform conserved functions in vertebrate and invertebrate striated and oblique muscles, although less is known about the structure of invertebrate connectins at present. The protein that maintains such a structure is present not only in vertebrate striated muscle, but also in invertebrate striated and oblique muscle. In the present study, we analyzed the partial primary structure of a 1200K-protein, which is a connectin-like protein that is expressed in Neanthes sp. body wall muscle that is in turn composed of oblique muscle. Antibody screening of a cDNA library of Neanthes sp. body wall muscle identified two different clones. Both clones coded for a sequence predominantly comprised of the four amino acids proline (P), glutamate (E), valine (V) and lysine (K). One clone included a PEVK-like repeat sequence flanked by an Ig domain, while the other clone comprised a distinct 14 amino acid repeat rich in PEVK residues, flanked by a non-repetitive unique sequence. The PEVK region is found in vertebrate connectin and is thought to generate elasticity and be responsible for passive tension of the muscle. The antibodies produced against a portion of each clone both reacted with bands corresponding to 1200 kDa present in Neanthes sp. body wall muscle. Therefore, our results demonstrate that this 1200K-protein is a connectin-like elastic protein and includes specific PEVK-like fragment. We suggest that this 1200K-protein plays a major role in maintaining the structure of oblique muscle in invertebrates.

Amino Acid Sequence↗

Species and tissue distribution of cholecystokinin/gastrin-like substances in some invertebrates.

Twenty-six species of invertebrates representing eight phyla were surveyed for the presence of cholecystokinin/gastrin-like (CCK/gastrin-like) peptides by radioimmunoassay of various tissue extracts. This is the first report of the presence of CCK/gastrin-like peptides in representatives of the phylum Ectoprocta, the arthropodan classes Crustacea and Merostomata, and in the nervous systems of the gastropod mollusc Aplysia californica and the oligochaete annelid Lumbricus terrestris. It has been proposed that CCK/gastrin evolved in the invertebrates as a neural peptide and was subsequently exploited by the vertebrates as a regulatory peptide in both the nervous system and the gastrointestinal endocrine system. The present results indicate that some gastropod molluscs, a merostomatan arthropod, and an annelid have detectable CCK/gastrin in both nervous and gut tissue. However, extractable CCK/gastrin was found only in gut tissue and not in the central nervous system of a crustacean arthropod. The tissue origin of the extracted CCK/gastrin in Bugula (phylum Ectoprocta) was not determined. Final resolution of the question of the nervous versus gut endocrine cellular origin of CCK/gastrin in invertebrates awaits further investigation. CCK/gastrin-like peptides are widely distributed among the invertebrates, which thus provide a rich source of comparative material for study of these regulatory substances.

Animals↗

Purification of invertebrate metallothioneins.

Purification procedures that appear to be generally applicable to invertebrate MTs have only recently been developed and are described here. Thus far, few invertebrate MTs have been purified and characterized, although proteins that exhibit similarities to MT have been identified in numerous invertebrate species. A greater understanding of MTs of this group, which comprises most of the animals in the animal kingdom, will provide the basis for increased understanding of the evolution of this ubiquitous protein. Additionally, specific invertebrate species may provide useful models for probing MT function and contribute to a greater understanding of the biological role of MT.

Animals↗

Mechanism of acute silver toxicity in marine invertebrates.

In freshwater crustaceans and in both freshwater and marine fish, the key mechanism of acute silver toxicity involves ionoregulatory impairment. An inhibition of the Na+ ,K+-ATPase located at the basolateral membrane of the gill epithelium seems to be the key site for silver toxicity. However, studies to determine if the same mechanism of toxicity is occurring in marine invertebrates, which also are ionoregulators, had not been done. Thus, the present study was carried out to determine acute silver effects on hemolymph osmo- and ionoregulation in three marine invertebrates: the shrimp Penaeus duorarum, the sea hare Aplysia californica, and the sea urchin Diadema antillarum. Animals were exposed to silver (1 or 10 microg/L), as silver nitrate, in seawater for 48 h. Results show that acute silver exposure did not affect hemolymph osmolality or ion concentration (Na+, Cl-, K+, Ca2+ and Mg2+) in the three species studied. However, silver induced significant changes in the water content in shrimp gill and sea hare gill and hepatopancreas. Silver also caused significant changes in Na+ ,K+-ATPase activity and in both total and intracellular ion (Cl-, Na+, K+, Mg2+, and Ca2+) concentrations in different tissues of the three species studied. Overall, these results show that the key mechanism of acute silver toxicity in marine invertebrates is not associated with an osmotic or ionoregulatory impairment at the hemolymph level, as observed in freshwater fish and crustaceans and in seawater fish. However, they indicate that acute waterborne silver induces significant changes in Na+ ,K(+)-ATPase activity and probably affects other mechanisms involved in water and ion transport at the cell membrane level, inducing impairments in water and ion regulation at the cellular level in different tissues of marine invertebrates. These results indicate the need to consider other "toxic sites" than gills in any future extension of the biotic ligand model (BLM) for seawater.

Analysis of Variance↗

Nitrate toxicity to aquatic animals: a review with new data for freshwater invertebrates.

Published data on nitrate (NO3-) toxicity to freshwater and marine animals are reviewed. New data on nitrate toxicity to the freshwater invertebrates Eulimnogammarus toletanus, Echinogammarus echinosetosus and Hydropsyche exocellata are also presented. The main toxic action of nitrate is due to the conversion of oxygen-carrying pigments to forms that are incapable of carrying oxygen. Nitrate toxicity to aquatic animals increases with increasing nitrate concentrations and exposure times. In contrast, nitrate toxicity may decrease with increasing body size, water salinity, and environmental adaptation. Freshwater animals appear to be more sensitive to nitrate than marine animals. A nitrate concentration of 10 mg NO3-N/l (USA federal maximum level for drinking water) can adversely affect, at least during long-term exposures, freshwater invertebrates (E. toletanus, E. echinosetosus, Cheumatopsyche pettiti, Hydropsyche occidentalis), fishes (Oncorhynchus mykiss, Oncorhynchus tshawytscha, Salmo clarki), and amphibians (Pseudacris triseriata, Rana pipiens, Rana temporaria, Bufo bufo). Safe levels below this nitrate concentration are recommended to protect sensitive freshwater animals from nitrate pollution. Furthermore, a maximum level of 2 mg NO3-N/l would be appropriate for protecting the most sensitive freshwater species. In the case of marine animals, a maximum level of 20 mg NO3-N/l may in general be acceptable. However, early developmental stages of some marine invertebrates, that are well adapted to low nitrate concentrations, may be so susceptible to nitrate as sensitive freshwater invertebrates.

Amphibians↗

Genetics of anti-parasite resistance in invertebrates.

This review summarizes and compares available data on genetic and molecular aspects of resistance in four well-described invertebrate host-parasite systems: snail-schistosome, mosquito-malaria, mosquito-filarial worm, and Drosophila-wasp associations. It underlies that the major components of the immune reaction, such as hemocyte proliferation and/or activation, and production of cytotoxic radicals are common to invertebrate hosts. Identifying genes responsible for naturally occurring resistance will then be helpful to understand the mechanisms of invertebrate immune defenses and to determine how virulence factors are used by parasites to overcome host resistance. Based on these four well-studied models, invertebrate resistance appears as generally determined by one major locus or a few loci, displaying at least partial dominance. Interestingly, specificity of resistance is highly variable and would involve processes other than simple recognition mechanisms. Finally, resistance was shown to be generally costly but is nevertheless observed at high frequencies in many natural populations, suggesting a high potential for host parasite coevolution.

Animals↗

Invertebrates as animal models for Staphylococcus aureus pathogenesis: a window into host-pathogen interaction.

Recently, the use of invertebrate models of infection has given exciting insights into host-pathogen interaction for a number of bacteria. In particular, this has revealed important factors of the host response with remarkable parallels in higher organisms. Here, we review the advances attained in the elucidation of virulence determinants of a major human pathogen, Staphylococcus aureus, in relation to the invertebrate models thus far applied, the silkworm (Bombyx mori), the fruit fly (Drosophila melanogaster) and the roundworm (Caenorhabditis elegans). Also, the major pathways of host defence are covered in light of the response to S. aureus and the similarities and divergences in innate immunity of vertebrates and invertebrates. Consequently, we comparatively consider pathogen recognition receptors, signal transduction pathways (including Toll, Imd and others), and the humoral and cellular antimicrobial effectors. The technically convenient and ethically acceptable invertebrates appear as a valuable first tool to discriminate molecules participating from both sides of the host-S. aureus interaction as well as a high throughput method for antimicrobial screening.

Animals↗

History of the Society for Invertebrate Pathology.

Scientists studying diseases of invertebrates in the USA, Europe, and Asia began to meet at international congresses in the 1950s and early 1960s, and soon recognized that they needed both a society and a journal where their common interests could be discussed and their findings presented. Edward A. Steinhaus played a major role in bringing together scientists from across the globe with common interests in these diseases. As a consequence, the Journal of Invertebrate Pathology (then Journal of Insect Pathology) was initiated in 1959 and Steinhaus became its first editor. Along with Albert Sparks he organized a meeting at Seattle, Washington in 1967 that led to the founding of the Society for Invertebrate Pathology with Steinhaus as its first President. The Society held its first meeting at Ohio State University in 1968, and has continued to meet annually. The Society has instituted a Founder's Lecture series, graduate student awards, and Divisions of Microbial Control, Microsporidia, Bacteriology, Fungi, Viruses, and Nematodes. Members enjoy several social functions at meetings as well as symposia, submitted papers, and poster sessions. The Society for Invertebrate Pathology is a truly international organization which to date has held meetings in 13 countries and 14 US states, usually attended by members from at least 20 countries.

Animals↗

Comparative approaches to facilitate the discovery of prolongevity interventions: effects of tocopherols on lifespan of three invertebrate species.

Many compounds hold promise for pharmacologic manipulation of aging. However, such claims are difficult to investigate due to time and budget constraints. Here, we took a comparative approach, using short-lived invertebrate species, to directly test the effects of two tocopherols (Vitamin E) on longevity. gamma-Tocopherol represents the most abundant tocopherol in the Western diet, while alpha-tocopherol is selectively enriched in human plasma. Both isoforms demonstrate antioxidant activity and are proposed to have anti-aging activities. We compared the effects of alpha- and gamma-tocopherol supplementation on lifespan in three invertebrate species. gamma-Tocopherol, but not alpha-tocopherol, slightly extended lifespan in nematodes, but neither significantly affected lifespan in two fly species. This study shows that a comparative approach, utilizing multiple invertebrate species, can increase the robustness of invertebrate-based pilot screens for prolongevity interventions.

Animals↗

Neuro-endocrine biomarkers of pollution-induced stress in marine invertebrates.

Nitric oxide (NO) is a molecular mediator of the non-specific immune response of vertebrates but production has also been recorded in response to bacterial infection in invertebrates. Together with biogenic amines and cytokines, production of NO in invertebrate phagocytic haemocytes is thought to represent a putative stress response. In this study the hypothesis that NO might be a sensitive biomarker of stress in invertebrates has been tested. The spectrophotometric Griess assay for nitrite and nitrate has been modified so that the final breakdown products of NO are suitable for measurement in invertebrate haemolymph. The assay was linear in the range 1-100 microM with a sensitivity of 4 microM. The baseline NOx measurement in unstressed mussels was 0.74 mM NOx/mg protein. Mytilus edulis were kept at 15 micro C in filtered seawater for at least 2 days prior to exposure to TBTO (0.001-0.1 mg/l) for 24 h. Preliminary results suggest that TBTO perturbs the NO response and that the outlined assay protocol is a sensitive means of detecting those changes. It is proposed that NO measurements potentially offer a highly sensitive, non-invasive means of monitoring stress responses associated with environmental change.

Adaptation, Physiological↗

Cell adhesion molecules in invertebrate immunity.

Cell adhesion is essential in immunity in invertebrates, e.g., in the cellular immune responses of encapsulation and nodule formation. Here cell adhesion molecules shown or suggested to be involved in invertebrate immunity are reviewed. Blood cells of the crayfish, Pacifastacus leniusculus, can release a cell-adhesive and opsonic peroxidase, peroxinectin. A site containing the motif, KGD, appears to be adhesive by binding to a transmembrane receptor of the integrin family on the blood cells. Peroxinectin also binds a peripheral blood cell surface CuZn-superoxide dismutase. The peroxidase-integrin interaction appears to have evolved early and seems conserved; human myeloperoxidase supports cell adhesion via the alphaMbeta2 integrin. There is evidence for peroxinectin-like proteins in other arthropods. Effects by RGD peptides indicate that integrins mediate blood cell adhesion and cellular immunity in diverse invertebrate species. Other invertebrate blood cell molecules proposed to be involved in adhesion include the insect plasmatocyte-spreading peptide, as well as soluble and transmembrane proteins which show some similarity to vertebrate adhesive or extracellular matrix molecules. Proteins such as the Ig family member hemolin, or proteins found in insects that are hosts for parasitic wasps, inhibit cell adhesion and may regulate or block cellular immunity.

Animals↗

Identification of endocrine-disrupting effects in aquatic vertebrates and invertebrates: report from the European IDEA project.

The EU-funded project IDEA aimed to evaluate (a) what parameters and endpoints allow the detection of endocrine-mediated developmental and reproductive effects of (xeno)estrogens in life cycle- and life stage-specific toxicity tests with the zebrafish Danio rerio, a small laboratory fish used in many ecotoxicity test guidelines, and (b) whether substances that act as estrogens in vertebrates may also adversely affect the development, differentiation, and reproduction of aquatic invertebrates. The invertebrate species investigated included Hydra vulgaris, Gammarus pulex, Chironomus riparius, Hyalella azteca, and Lymnaea stagnalis. The animals were exposed to the model estrogenic chemicals ethynylestradiol (EE2), bisphenol A (BPA), and octylphenol (OP), which exert their endocrine activity in vertebrates through the estrogen receptor. As endpoints, developmental and reproductive parameters at the organism level as well as molecular and cellular parameters were measured. Life cycle exposure of zebrafish to (xeno)estrogens induced a specific, partly irreversible response pattern, consisting mainly of (a) induction of vitellogenin (VTG), (b) alterations of gonad differentiation, (c) delay of first spawning, and (d) reduced fertilization success. The effects of EE2 on zebrafish were expressed at environmentally realistic concentrations, while BPA and OP became effective at concentrations higher than those usually found in the environment. The vitellogenic response was equally sensitive as the reproductive parameters in the case of EE2, but VTG was more sensitive in the case of BPA. Partial life cycle exposure of zebrafish had lasting effects on fish development and reproduction only when the fish were exposed during the stage of juvenile bisexual gonad differentiation. In (partial) life cycle and multigeneration studies with invertebrates, (xeno)estrogenic impact was assessed by a range of developmental and reproductive parameters including hatching, growth, moulting, mating behavior, and egg number. Several parameters were found to be responsive to (xeno)estrogens; however, most effects were induced only at higher, probably nonphysiological concentrations. Low-dose effects were observed in full life cycle experiments, particularly in the second generation. It remains to be established whether the estrogen-induced alterations in the invertebrate species indeed do result from disturbances of the endocrine system. The findings of the present research project support the development of appropriate testing methodologies for substances with estrogenic activity.

Animals↗

Polychlorinated biphenyl accumulation differs among pumpkinseed sunfish during experimental field exposure: the role of invertebrate prey.

The relative importance of aqueous vs. trophic exposure of fish to polychlorinated biphenyls (PCBs) was investigated. The potential role of different invertebrate prey upon PCB accumulation by fish was also investigated. Pumpkinseed sunfish (Lepomis gibbosus) were exposed in the upper Hudson River under conditions that either permitted feeding upon different local prey assemblages, or excluded local prey from fish diets. Total [PCB] was 5x greater in fish exposed to local prey versus those not permitted to feed on local prey. Further, fish with trophic exposure to PCBs exhibited a chlorine shift, a significantly higher proportion of more chlorinated isomers than fish with only aqueous exposure. Total [PCB] differed among benthic invertebrate assemblages. Phytophilous invertebrates had lower [PCB] than benthic invertebrates, and also had lower concentrations of the more chlorinated isomers. Short-term (7-day) exposure of fish did not permit differentiation of PCB accumulation in fish according to differences in diet of native prey assemblages.

Animals↗