Lamprey 48-kDa lens protein represents a novel class of crystallins

SDS-PAGE revealed a major Mr 48 000 polypeptide of pI around 8 in the water-soluble fraction of lamprey lenses. It occurs as a monomeric protein, and its amino acid composition and tryptic peptides show no resemblances to alpha-, beta-, gamma- or delta-crystallin. Immunoblotting with antiserum against the 48-kDa protein revealed an immunologically related polypeptide of similar Mr in reptiles, several birds and a fish, but showed no cross-reactivity with any other water-soluble lens component. The 48-kDa protein is not detected in many birds and fishes, and in the investigated mammals and amphibians.     

Myelin tetraspan family proteins but no non-tetraspan family proteins are present in the ascidian (Ciona intestinalis) genome

Several of the proteins used to form and maintain myelin sheaths in the central nervous system (CNS) and the peripheral nervous system (PNS) are shared among different vertebrate classes. These proteins include one-to-several alternatively spliced myelin basic protein (MBP) isoforms in all sheaths, proteolipid protein (PLP) and DM20 (except in amphibians) in tetrapod CNS sheaths, and one or two protein zero (P0) isoforms in fish CNS and in all vertebrate PNS sheaths. Several other proteins, including 2', 3'-cyclic nucleotide 3'-phosphodiesterase (CNP), myelin and lymphocyte protein (MAL), plasmolipin, and peripheral myelin protein 22 (PMP22; prominent in PNS myelin), are localized to myelin and myelin-associated membranes, though class distributions are less well studied. Databases with known and identified sequences of these proteins from cartilaginous and teleost fishes, amphibians, reptiles, birds, and mammals were prepared and used to search for potential homologs in the basal vertebrate, Ciona intestinalis. Homologs of lipophilin proteins, MAL/plasmolipin, and PMP22 were identified in the Ciona genome. In contrast, no MBP, P0, or CNP homologs were found. These studies provide a framework for understanding how myelin proteins were recruited during evolution and how structural adaptations enabled them to play key roles in myelination.     

Some introductory notes on the organization of the forebrain in tetrapods.

As an introduction to the main theme of this conference an overview of the organization of the tetrapod forebrain is presented with emphasis on the telencephalic representation of sensory and motor functions. In all classes of tetrapods, olfactory, visual, octavolateral, somatosensory and gustatory information reaches the telencephalon. Major differences exist in the telencephalic targets of sensory information between amphibians and amniotes. In amphibians, three targets are found: the lateral pallium for olfactory input, the medial pallium for visual and multisensory input, and the lateral subpallium for visual, octavolateral and somatosensory information. The forebrains of reptiles and mammals are similar in that the dorsal surface of their cerebral hemisphere is formed by a pallium with three major segments: (a) an olfactory, lateral cortex; (b) a 'limbic' cortex that forms the dorsomedial wall of the hemisphere, and (c) an intermediate cortex that is composed entirely of isocortex in mammals, but in reptiles (and birds) consists of at least part of the dorsal cortex (in birds the Wulst) and a large intraventricular protrusion, i.e. the dorsal ventricular ridge. In birds, the entire lateral wall of the hemisphere is involved in this expansion. The intermediate pallial segment receives sensory projections from the thalamus and contains modality-specific sensory areas in reptiles, birds and mammals. The most important differences between the intermediate pallial segment of amniotes concern motor systems.     

Phylogenetic Survey of Proteins Related to Synapsin I and Biochemical Analysis of Four Such Proteins from Fish Brain

A phylogenetic survey of proteins immunologically related to Synapsin I, a major synaptic vesicle-associated phosphoprotein in mammals was carried out. Proteins antigenically related to Synapsin I were found by use of radioimmunoassay and other radioimmunochemical techniques in the nervous systems of several vertebrate and invertebrate species, which included birds, reptiles, amphibians, fish, echinoderms, arthropods, and mollusks. Four proteins present in fish brain, antigenically related to Synapsin I, were further studied and found to resemble mammalian Synapsin I in several respects. Like Synapsin I, the fish proteins were present in high amounts in nervous tissue, were enriched in synaptosomal fractions of brain where they were substrates for endogenous protein kinases, were acid extractable, and were sensitive to digestion by collagenase. In addition, two-dimensional peptide-mapping analysis revealed some homology between major phosphopeptide fragments of Synapsin I and the fish proteins. The results indicate that proteins related to Synapsin I are wide-spread in the animal kingdom.     

Structure,activity, and distribution of fish osteocalcin

Osteocalcin (bone Gla protein) is an extracellular matrix protein synthesized by osteoblasts that is a marker of bone. Osteocalcin probably originated in the ancestors of Teleostei or bony fish and of the Tetrapoda or amphibians, reptiles, birds, and mammals. We have characterized the Cyprinus carpio (carp) osteocalcin for mineral binding to hydroxyapatite, amino acid sequence, and extent of secondary structure. Hydroxyapatite binding is enhanced in the presence of calcium. The alpha-helical content of teleost osteocalcin increases and beta-sheet structure decreases upon calcium binding, similar to findings in calf osteocalcin. The gene structure and primary sequence of prepro-osteocalcin from 2 pufferfish compared with carp shows that there are many conserved features in teleost osteocalcin genes. Using an immunoassay for carp osteocalcin, we determined that the relative content of osteocalcin is highest in dorsal fin spines and other bones and lowest in scales. The carp osteocalcin antibodies, cross-reactive to other species of fish, were used to study the role of osteocalcin in teleost model systems.     

Fish models in prion biology: Underwater issues

Transmissible spongiform encephalopathies (TSEs), otherwise known as prion disorders, are fatal diseases causing neurodegeneration in a wide range of mammalian hosts, including humans. The causative agents - prions - are thought to be composed of a rogue isoform of the endogenous prion protein (PrP). Beyond these and other basic concepts, fundamental questions in prion biology remain unanswered, such as the physiological function of PrP, the molecular mechanisms underlying prion pathogenesis, and the origin of prions. To date, the occurrence of TSEs in lower vertebrates like fish and birds has received only limited attention, despite the fact that these animals possess bona fide PrPs. Recent findings, however, have brought fish before the footlights of prion research. Fish models are beginning to provide useful insights into the roles of PrP in health and disease, as well as the potential risk of prion transmission between fish and mammals. Although still in its infancy, the use of fish models in TSE research could significantly improve our basic understanding of prion diseases, and also help anticipate risks to public health. This article is part of a Special Issue entitled Zebrafish Models of Neurological Diseases.     

Analysis of 27 mammalian and 9 avian PrPs reveals high conservation of flexible regions of the prion protein.

Prion diseases are fatal neurodegenerative disorders in man and animal associated with conformational conversion of a cellular prion protein (PrPc) into the pathologic isoform (PrPSc). The function of PrPcand the tertiary structure of PrPScare unclear. Various data indicate which parts of PrP might control the species barrier in prion diseases and the binding of putative factors to PrP. To elucidate these features, we analyzed the evolutionary conservation of the prion protein. Here, we add the primary PrP structures of 20 ungulates, three rodents, three carnivores, one maritime mammal, and nine birds. Within mammals and birds we found a high level of amino acid sequence identity, whereas between birds and mammals the overall homology was low. Various structural elements were conserved between mammals and birds. Using the CONRAD space-scale alignment, which predicts conserved and variable blocks, we observed similar patterns in avian and mammalian PrPs, although 130 million years of separate evolution lie in between. Our data support the suggestion that the repeat elements might have expanded differently within the various classes of vertebrates. Of note is the N-terminal part of PrP (amino acid residues 23-90), which harbors insertions and deletions, whereas in the C-terminal portion (91-231) mainly point mutations are found. Strikingly, we found a high level of conservation of sequences that are not part of the structured segment 121-231 of PrPcand of the structural elements therein, e.g. the N-terminal region from amino acid residue 23-90 and the regions located upstream of alpha-helices 1 and 3.     

两栖和爬行类生长及生长激素分泌活动的调节

根据生长内分泌学,综述了近年来两栖和爬行类生长激素（ＧＨ）分泌活动的调节及生长激素在两栖爬行类生长中作用这一研究领域所取得的主要成就和研究进展,研究结果表明：在脊椎动物的进化过程中,ＧＨ的化学结构和功能是相当保守的;ＧＨ对两栖爬行类的生长起促进作用;类胰岛素生长因子（ＩＧＦｓ）也能冰分传递ＧＨ促进两栖类的生长;在两栖爬行类,下丘脑对ＧＨ分泌的调控较哺乳类缺乏特异性,利用外源ＧＨ促进两栖爬行类的生长     

Structural characteristics of genome organization in amphibians: Differential staining of chromosomes and DNA structure

Summary Differential staining patterns on amphibian chromosomes are in some respects distinct from those on mammalian chromosomes; C-bands are best obtained, whereas G- and Q-bands are either unobtainable (on anuran chromosomes) or coincide with C-bands (chromosomes of urodeles). In amphibians, rRNA genes are located at secondary constrictions, but in urodeles they are also found at other chromosome sites, the positions of these sites being strictly heritable. DNA content in amphibian cells is tens and hundreds times higher than in mammals. DNA contents in anurans and urodeles differ within certain limits: from 2 to 25 pg/N and from 30 to over 160 pg/N respectively. Species characterized by slow morphogenesis have larger genomes. Genome growth is normally due to an increase in the amount of repetitive DNA (mostly intermediate repetitive sequences), the amount of unique sequences being almost constant (11 pg/genome in urodeles, and 1.5 pg/genome in anurans). In anurans in general no satellite DNA was found, whereas such fractions were found in manyUrodela species. Nucleosome chromatin structure in amphibians is identical to that of other eukariotes. It is postulated that differences in chromosome banding between amphibians and mammals are due to differences in chromatin packing which in turn is related to the distinct organization of DNA repetitive sequences. It is likely that fish chromosomes have a similiar structure. A comparison of such properties as the chromosome banding patterns, variations in nuclear DNA content and some genome characteristics enable us to group fishes and amphibians together as regards chromosome structure, as distinct from amniotes - reptiles, birds and mammals. It is probable that in the ancient amphibians - ancestors of reptiles - chromatin packing underwent a radical transformation, following changes in the organization of DNA repetitive sequences.     

Distribution of immunoreactive Tamm-Horsfall protein in various species in the vertebrate classes

A sheep antibody to human Tamm-Horsfall protein, the major protein in normal urine, was used in an immunohistological study of organs of 48 species of vertebrate animals, representing the classes Mammalia, Aves, Reptilia, Amphibia, Osteichthyes and Chondrichthyes. Immunoreacvity was shown in the thick limb of the loop of Henle in the kidney of mammals, but there was no reactivity with tissues of birds or reptiles. Superficial layers of the skin of several amphibians and fish, superficial layers of the oral mucosa and gills of fish, and the distal tubules of the kidney of some amphibians, reacted with the antibody. Immunoreactivity with mammalian kidney was removed by passage of the antibody down an immunoadsorption column coated with human Tamm-Horsfall protein, and amphibian immunoreactivity was removed by incubation of the antibody with material prepared from frogs in the same way as Tamm-Horsfall protein. These findings suggest that immunoreactive Tamm-Horsfall protein appeared early in vertebrate phylogeny, initially in skin and gills and later in kidney, and that although conserved in evolution, it shows antigenic differences between amphibians and mammals. Its distribution is consistent with the hypothesis that it acts as a waterproofing agent.     

Prion protein-related proteins from zebrafish are complex glycosylated and contain a glycosylphosphatidylinositol anchor

A hallmark of prion diseases in mammals is a conformational transition of the cellular prion protein (PrP(C)) into a pathogenic isoform termed PrP(Sc). PrP(C) is highly conserved in mammals, moreover, genes of PrP-related proteins have been recently identified in fish. While there is only little sequence homology to mammalian PrP, PrP-related fish proteins were predicted to be modified with N-linked glycans and a C-terminal glycosylphosphatidylinositol (GPI) anchor. We biochemically characterized two PrP-related proteins from zebrafish in cultured cells and show that both zePrP1 and zeSho2 are imported into the endoplasmic reticulum and are post-translationally modified with complex glycans and a C-terminal GPI anchor.     

Relationships among nutrition and reproduction and relevance for wild animals

This review discusses aspects of feeding ecology, nutrition, and dietary husbandry that are particularly relevant to reproductive success in wild animals. Emphasis has been placed on recently published literature. Special attention has been given to requirements for energy and protein and the unique roles of essential amino acids, essential fatty acids, calcium, phosphorus, zinc, copper, selenium, and vitamins A, C, D, and E. Information has been drawn from research with mammals, birds, reptiles, amphibians, and fish, including the domestic and laboratory species that play such a large role in elucidating mechanisms connecting nutrition and reproduction. Zoo Biol 23:475–487, 2004. © 2004 Wiley-Liss, Inc.     

The diet of feral cats on islands: a review and a call for more studies

Cats are among the most successful and damaging invaders on islands and a significant driver of extinction and endangerment. Better understanding of their ecology can improve effective management actions such as eradication. We reviewed 72 studies of insular feral cat diet from 40 islands worldwide. Cats fed on a wide range of species from large birds and medium sized mammals to small insects with at least 248 species consumed (27 mammals, 113 birds, 34 reptiles, 3 amphibians, 2 fish and 69 invertebrates). Three mammals, 29 birds and 3 reptiles recorded in the diet of cats are listed as threatened by the IUCN. However, a few species of introduced mammals were the most frequent prey, and on almost all islands mammals and birds contributed most of the daily food intake. Latitude was positively correlated with the predation of rabbits and negatively with the predation of reptiles and invertebrates. Distance from landmass was positively correlated with predation on birds and negatively correlated with the predation of reptiles. The broad range of taxa consumed by feral cats on islands suggests that they have the potential to impact almost any native species, even the smallest ones under several grams, that lack behavioral, morphological or life history adaptations to mammalian predators. Insular feral cat??s reliance on introduced mammals, which evolved with cat predation, suggests that on many islands, populations of native species have already been reduced.     

Transthyretin (prealbumin) gene expression in choroid plexus is strongly conserved during evolution of vertebrates

1. The major protein synthesized and secreted by the choroid plexus from mammals, birds, reptiles and probably amphibians is similar in subunit structure to transthyretin. 2. In mammals and birds the proportion of transthyretin mRNA is much higher in choroid plexus RNA than in liver RNA. No transthyretin mRNA is found in brain outside the choroid plexus. 3. Transthyretin-like protein, such as that secreted by the choroid plexus, was not detected in amphibian serum and was present in very low levels in reptile serum. 4. It is proposed that transthyretin synthesis and secretion arose earlier in evolution in the choroid plexus than in the liver.     

Body cavities in fishes

Four types of body cavity arrangement were found in 20 species of fish (see Materials and Methods). The body cavities of fishes were also compared with those of mammals and birds. It was concluded that higher vertebrates utilized only the Tinca type for their evolution, while the other types were dead-end evolutionary forms which terminated at fish level, or possibly with amphibians and reptiles. The necessary research in these groups still remains to be done.     

Molecular dynamics simulations of human prion protein: importance of correct treatment of electrostatic interactions.

Molecular dynamics simulations have been used to investigate the dynamical and structural behavior of a homology model of human prion protein HuPrP(90-230) generated from the NMR structure of the Syrian hamster prion protein ShPrP(90-231) and of ShPrP(<90-231) itself. These PrPs have a large number of charged residues on the protein surface. At the simulation pH 7, HuPrP(90-230) has a net charge of -1 eu from 15 positively and 14 negatively charged residues. Simulations for both PrPs, using the AMBER94 force field in a periodic box model with explicit water molecules, showed high sensitivity to the correct treatment of the electrostatic interactions. Highly unstable behavior of the structured region of the PrPs (127-230) was found using the truncation method, and stable trajectories could be achieved only by including all the long-range electrostatic interactions using the particle mesh Ewald (PME) method. The instability using the truncation method could not be reduced by adding sodium and chloride ions nor by replacing some of the sodium ions with calcium ions. The PME simulations showed, in accordance with NMR experiments with ShPrP and mouse PrP, a flexibly disordered N-terminal part, PrP(90-126), and a structured C-terminal part, PrP(127-230), which includes three alpha-helices and a short antiparallel beta-strand. The simulations showed some tendency for the highly conserved hydrophobic segment PrP(112-131) to adopt an alpha-helical conformation and for helix C to split at residues 212-213, a known disease-associated mutation site (Q212P). Three highly occupied salt bridges could be identified (E146/D144<-->R208, R164<-->D178, and R156<-->E196) which appear to be important for the stability of PrP by linking the stable main structured core (helices B and C) with the more flexible structured part (helix A and strands A and B). Two of these salt bridges involve disease-associated mutations (R208H and D178N). Decreased PrP stability shown by protein unfolding experiments on mutants of these residues and guanidinium chloride or temperature-induced unfolding studies indicating reduced stability at low pH are consistent with stabilization by salt bridges. The fact that electrostatic interactions, in general, and salt bridges, in particular, appear to play an important role in PrP stability has implications for PrP structure and stability at different pHs it may encounter physiologically during normal or abnormal recycling from the pH neutral membrane surface into endosomes or lysomes (acidic pHs) or in NMR experiments (5.2 for ShPrP and 4.5 for mouse PrP).     

Ferritins in Chordata: Potential evolutionary trajectory marked by discrete selective pressures

Ferritins (FTs) are iron storage proteins that are involved in managing iron‐oxygen balance. In our work, we present a hypothesis on the putative effect of geological changes that have affected the evolution and radiation of ferritin proteins. Based on sequence analysis and phylogeny reconstruction, we hypothesize that two significant factors have been involved in the evolution of ferritin proteins: fluctuations of atmospheric oxygen concentrations, altering redox potential, and changing availability of water rich in bioavailable ferric ions. Fish, ancient amphibians, reptiles, and placental mammals developed the broadest repertoire of singular FTs, attributable to embryonic growth in aquatic environments containing low oxygen levels and abundant forms of soluble iron. In contrast, oviparous land vertebrates, like reptiles and birds, that have developed in high oxygen levels and limited levels of environmental Fe²⁺ exhibit a lower diversity of singular FTs, but display a broad repertoire of subfamilies, particularly notable in early reptiles.     

Polypeptide YY- and neuropeptide Y-immunoreactive cells and nerves in the endocrine and exocrine pancreas of some vertebrates: An onto- and phylogenetic study

Summary The occurrence of polypeptide YY- and neuropeptide Y-immunoreactive cells and nerves in the pancreas of some species from all the eight main vertebrate groups (cyclostomes, cartilaginous fish, bony fish, amphibia, reptiles, birds, and mammals) was investigated. In addition, an ontogenetic study of these neurohormonal peptides was performed, using the rat pancreas. The distribution of these two peptides was compared with that of the structurally closely related pancreatic polypeptide.Polypeptide YY-immunoreactive cells were found to occur in the endocrine pancreas and neuropeptide Y-immunoreactivity was observed both in neurons and nerve fibres. The polypeptide YY-immunoreactive cells were limited to mammals and reptiles only. Neuropeptide Y-immunoreactive neurons and nerves were observed in reptiles, birds, and mammals only. One reptilian species (out of three) and one mammalian (out of six) failed to show any kind of immunoreactivity for the polypeptide or neuropeptide. Pancreatic polypeptide-immunoreactive cells were found in all the species examined except in the hagfish islet.In rat foetuses, polypeptide YY-immunoreactive cells and neuropeptide Y-immunoreactive nerve elements were first demonstrated at the seventeenth day of gestation, whereas pancreactic peptide-immunoreactive cells did not appear until postnatally, namely in two day-old rats. The polypeptide-containing cells, a new cell type in the endocrine pancreas, are rare. In contrast to the pancreatic peptide cells, they do not seem to have any kind of regional distribution.     

Characterization of the novel human transmembrane protein 9 (TMEM9) that localizes to lysosomes and late endosomes

We report the isolation and characterization of a cDNA coding for Fugu rubripes prion protein (PrP)-like of 180 amino acids which includes the PrP-conserved hydrophobic region homologous to that of Xenopus PrP. In addition to the hydrophobic region, Fugu PrP-like has several features common to PrPs, such as a signal sequence, a basic nature (pI 9.7) and a single intron in the 5' untranslated region. A possible glycosyl phosphatidylinositol (GPI) anchor site also exists in PrP-like. In expression analysis, PrP-like mRNA was detected in retina, skin, and brain, all of which express PrP mRNA in mammals. In a genome fragment clone (T002589, 31945 bp) sequenced by the Fugu Genomics Project, PrP-like located between KIAA0168 and SLC231A homologues. In human chromosome 20p13, PrP, Doppel, KIAA0168, and SLC231A align in this order. The close gene arrangement between the Fugu and human genomes suggests that Fugu PrP-like is a real orthologue of human PrP. However, Fugu PrP-like does not possess tandem repeats or a region with two glycosylation sites and a disulphide bridge. We do not declare that the cloned Fugu PrP-like represents fish PrP due to structural inconsistency, but believe that it will offer new insights into the evolution of PrPs from fish to tetrapods.