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PubMed1983

Physiological and biochemical properties and population studies in the M lactate dehydrogenase polymorphism of Discoglossus pictus (Amphibia, Anura).

1. M LDH polymorphism has been previously described in Discoglossus pictus. 2. Gene frequency at the LdM locus varied significantly in two isolated areas which differed markedly in water oxygen tension. 3. Sex distribution, relative activity of the LDH isozymes, percentage of total H and M subunits and enzyme kinetics did not differ among the distinct LDH phenotypes. 4. Under decreasing oxygen conditions, mean time to death, LT50 and oxygen consumption were similar in tadpoles of the three LDH phenotypes (fast, intermediate and slow). 5. Our results are compatible with a neutral role for this LDH polymorphism.

Animals↗
PubMed1983

Hepatic arginase activity in intra- and extrauterine larvae of the ovoviviparous salamander. Salamandra salamandra (L.) (Amphibia, Urodela).

The hepatic arginase activity of Salamandra salamandra was determined at three different stages of intra- and extrauterine larval development and at fully metamorphosed juveniles. The highest enzymatic activity was found in intrauterine larvae in November, the lowest in intrauterine larvae in June of the following year. These data can be correlated with the ureotelism of intrauterine larvae previously described and are discussed with respect to the metabolism of larval and juvenile fire salamanders.

Animals↗
PubMed1984

The distribution of monosaccharides and hexosamines in the oviduct of Salamandra salamandra (L.) (Amphibia, Urodela).

Biochemical analysis of the different segments of the oviduct in the ovoviviparous salamander, Salamandra salamandra, reveals the monosaccharides glucose, galactose, fucose, mannose, ribose and the hexosamines glucosamine and mannosamine. In segment 1 (pars recta) ribose and mannose are absent, and in segments 2 (p. convoluta I) and 5 (p. convoluta IV, uterus) mannose is not detectable; fucose is absent in the uterus. Segments 3 (p. convoluta II) and 4 (p. convoluta II) contain all sugars identified. The main hexoses present in the glandular segments are galactose, fucose and glucose.

Animals↗
PubMed1985

Yolk proteins and their plasmatic precursor in the tetraploid Odontophrynus americanus (Amphibia, Anura).

The vitellogenin of Odontophrynus americanus is a large (426.5 kDa) plasmatic protein. The vitellogenin is composed of two different phosphoglycopeptides: VTG1 = 207.5 kDa and VTG2 = 202.4 kDa. The vitellins originating from the partial proteolysis of the plasmatic vitellogenin on the ovary cells are composed of lipovitellins and phosphoproteins. Lipovitellin 1 has two glycopeptides with different amino acid sequences as determined by peptide mapping (LV1 alpha, 104.6 kDa; and LV1 beta, 92.6 kDa). Lipovitellin 2 is composed of three kinds of polypeptides (LV2 alpha, 31.7 kDa; LV2 beta, 29.7 kDa; LV2 gamma, 27.8 kDa). There are three phosphopeptides in the yolk: phosvitin (PV, 37.4 kDa) and phosvettes 1 (PVT1, 27.7 kDa) and 2 (PVT2, 26.1 kDa).

Animals↗
PubMed1994

Electrophoretic and morphometric evidence for two species of the genus Rana (Amphibia, Ranidae) and some aspects of their variabilities.

Frogs of two species, genus Rana, R. mascareniensis and R. levantina from northern Egypt, were examined using electrophoresis for 15 presumptive loci and measuring of 11 morphological characters, employing discriminate and principle component analysis. Electrophoretic and morphometric data showed clear interspecific differences. Using serum and tissue electrophoresis all individuals of R. mascareniensis had no intraspecific variabilities but R. levantina showed some intraspecific variabilities.

Alleles↗
PubMed1996

Identification of immunoreactive mammalian gonadotropin-releasing hormone in the brain of metamorphic larvae of Bufo arenarum Hensel (Amphibia: Anura).

Gonadotropin-releasing hormone (GnRH) immunoreactivity in brain extracts of Bufo arenarum tadpoles were investigated by high-performance liquid chromatography, followed by radioimmunoassay analysis using two different antisera raised against different GnRH variants. Only one immunoreactive peak was identified, eluting in the same position as synthetic mammalian GnRH. This result was further confirmed by serial dilution studies using more specific mammalian GnRH antisera. Our results suggest that mammalian GnRH is most likely an endogenous peptide in the brain of the developing larvae and froglets of Bufo arenarum and quite likely it is the only GnRH variant present during those development stages. The distribution and density of cell bodies and fibers were analysed by immunocytochemical procedures. Immunoreactive cell bodies appeared in the olfactory epithelium and across the olfactory nerve at late prometamorphic larval stages. Near the metamorphic climax and in froglets, perikarya and fibers were detected in basal forebrain, preoptic and hypothalamic areas. No immunoreaction was observed at midbrain, hindbrain and spinal cord levels. This study suggests that mammalian GnRH is most likely an endogenous peptide and is probably the only GnRH variant in the brain of the developing larvae and froglets of Bufo arenarum.

Animals↗
PubMed1994

Distribution of tyrosine hydroxylase immunoreactivity in the brain of Typhlonectes compressicauda (Amphibia, Gymnophiona): further assessment of primitive and derived traits of amphibian catecholamine systems.

Until now, catecholamine systems are well studied in the brains of anurans and urodeles, but such data are almost completely lacking for the third order of amphibians, i.e. the limbless Gymnophiona or Apoda. To further assess general and derived features of the catecholamine systems in this class of vertebrates, the distribution of tyrosine hydroxylase immunoreactive (THi) cell bodies and fibers was studied in the brain of the gymnophionan Typhlonectes compressicauda. The distribution of THi cell groups in the brain of gymnophionans largely resembles that found in anurans and urodeles. However, in gymnophionans additional THi cells were found in the reticular formation and in the prevagal part of the solitary tract nucleus. Other differences with anurans and urodeles concern the relatively larger number of THi cells in the midbrain tegmentum and in the hypothalamus, where the cells are mainly of the liquor-contacting type. The distribution of THi fibers in some brain regions of gymnophionans, e.g. pallial and basal forebrain areas, shows a greater resemblance with that of urodeles than with that of anurans. A peculiar feature of Typhlonectes are the pericellular baskets of THi varicosities in the lateral septal region. Such baskets were never observed in other amphibians, but do occur in the septal region of amniotes. Finally, the data obtained in this study support the suggestion that catecholamines play a role in the processing of sensory modalities such as olfactory, visual, auditory, vestibular, and mechanoreceptive lateral line information, but not in electroreception.

Amphibians↗
PubMed2003

Proton pump-driven cutaneous chloride uptake in anuran amphibia.

Krogh introduced the concept of active ion uptake across surface epithelia of freshwater animals, and proved independent transports of Na(+) and Cl(-) in anuran skin and fish gill. He suggested that the fluxes of Na(+) and Cl(-) involve exchanges with ions of similar charge. In the so-called Krogh model, Cl(-)/HCO(3)(-) and Na(+)/H(+) antiporters are located in the apical membrane of the osmoregulatory epithelium. More recent studies have shown that H(+) excretion in anuran skin is due to a V-ATPase in mitochondria-rich (MR) cells. The pump has been localized by immunostaining and H(+) fluxes estimated by pH-stat titration and mathematical modelling of pH-profiles in the unstirred layer on the external side of the epithelium. H(+) secretion is voltage-dependent, sensitive to carbonic-anhydrase inhibitors, and rheogenic with a charge/ion-flux ratio of unity. Cl(-) uptake from freshwater is saturating, voltage independent, and sensitive to DIDS and carbonic-anhydrase inhibitors. Depending on anuran species and probably on acid/base balance of the animal, apical exit of protons is coupled to an exchange of Cl(-) with base (HCO(3)(-)) either in the apical membrane (gamma-type of MR cell) or in the basolateral membrane (alpha-type MR cell). The gamma-cell model accounts for the rheogenic active uptake of Cl(-) observed in several anuran species. There is indirect evidence also for non-rheogenic active uptake accomplished by a beta-type MR cell with apical base secretion and basolateral proton pumping. Several studies have indicated that the transport modes of MR cells are regulated via ion- and acid/base balance of the animal, but the signalling mechanisms have not been investigated. Estimates of energy consumption by the H(+)-ATPase and the Na(+)/K(+)-ATPase indicate that the gamma-cell accomplishes uptake of NaCl in normal and diluted freshwater. Under common freshwater conditions with serosa-positive or zero V(t), the K(+) conductance of the basolateral membrane would have to maintain the inward driving force for Na(+) uptake across the apical membrane. With the K(+) equilibrium potential across the basolateral membrane estimated to -105 mV, this would apply to external Na(+) concentrations down to 40-120 micromol/l. NaCl uptake from concentrations down to 10 micromol/l, as observed by Krogh, presupposes that the H(+) pump hyperpolarizes the apical membrane, which would then have to be associated with serosa-negative V(t). In diluted freshwater, exchange of cellular HCO(3)(-) with external Cl(-) seems to be possible only if the proton pump has the additional function of keeping the external concentration of HCO(3)(-) low. Quantitative considerations also lead to the conclusion that with the above extreme demand, at physiological intracellular pH of 7.2, the influx of Cl(-) via the apical antiporter and the passive exit of Cl(-) via basolateral channels would be possible within a common range of intracellular Cl(-) concentrations.

Animals↗
PubMed2006

Novel dermaseptins from Phyllomedusa hypochondrialis (Amphibia).

Six new antimicrobial peptides structurally related to the dermaseptin family have been isolated from the skin secretion of the amphibian Phyllomedusa hypochondrialis. The primary structures of these molecules named as DShypo 01, 02, 03, 04, 06, and 07 were determined by de novo MS/MS experiments, Edman degradation, and cDNA sequencing. The fifth peptide was found to be precisely the same DS 01 from Phyllomedusa oreades previously described by our group. The majority of the peptides purified from the crude skin secretion could be directly localized and mapped onto a freshly dissected dorsal skin fragment using mass spectrometry-imaging techniques. Comparisons between peptides and commercial drugs on their antibacterial and anti-Leishmania amazonensis efficiencies, associated with peptide lytic effects on mammalian blood cells and surface plasmon resonance interaction studies on immobilized DMPC vesicles, were also performed.

Amino Acid Sequence↗
PubMed2004

A subterranean generalist predator: diet of the soil-dwelling caecilian Gegeneophis ramaswamii (Amphibia; Gymnophiona; Caeciliidae) in southern India.

Biologists have paid relatively little attention to subterranean predators, especially their ecology. Although diets of some subterranean lower vertebrates suggest specialisation, there remains a lack of quantitative data. The diet of the caecilian amphibian Gegeneophis ramaswamii was investigated through analyses of gut contents of 67 specimens collected in randomised surveys at three localities in Kerala, southern India, in early and mid-monsoon. Although termites were the most frequently ingested items in the mid-monsoon, the specialist predator hypothesis was rejected because of differences in diet found in early monsoon samples, when earthworms contributed the greatest mass. That guts of some G. ramaswamii contained many individuals of only a single dietary taxon was interpreted as feeding on patchily distributed prey rather than specialisation. No ontogenetic differences in diet were apparent, but more sampling is required to investigate this further. Subadults largely feed on fewer items of the same prey as adults, though there is an indication that subadult diet is less diverse. The data do not support differences between male and female diet. High densities of G. ramaswamii, and perhaps of other terrestrial caecilians and subterranean lower vertebrates feeding on soil-ecosystem engineers (termites, earthworms and ants), might substantially impact soil ecology.

Amphibians↗
PubMed2005

Well known does not mean well studied: morphological and molecular support for existence of sibling species in the Javanese gliding frog Rhacophorus reinwardtii (Amphibia, Anura).

Morphological, morphometrical and molecular data support the existence of two sibling species in the taxon previously known as Rhacophorus reinwardtii. The two species can be distinguished by the coloration pattern of webbing, the size of adult specimens and the relative size of various morphometric characters. This long and commonly known taxon should be separated into two species. As a consequence, the conservation status of the new species Rhacophorus kio n. sp. as well as of the redefined species Rhacophorus reinwardtii must be re-evaluated and, considering the new distribution data and the particular ecological demands, both species should be considered as 'endangered'.

Animals↗
PubMed2005

The complete mitochondrial genome of a tree frog, Polypedates megacephalus (Amphibia: Anura: Rhacophoridae), and a novel gene organization in living amphibians.

In this study, we have determined the complete sequence of the mitochondrial genome of an Old World tree frog Polypedates megacephalus (Anura: Rhacophoridae) by using a long polymerase chain reaction (PCR) technique and shotgun strategy of sequencing. The entire mtDNA sequence is 16,473 nt long with a novel mitogenomic gene organization in amphibians. Unlike other neobatrachian frogs, the transfer ribonucleic acid (tRNA)-Leu(CUN) and tRNA-Thr genes exchange their positions in P. megacephalus and form a Thr-Leu(CUN)-Pro-Phe tRNA gene tetrad. Moreover, we found that the ATP8 gene was replaced by a noncoding sequence of 853 nt long and that the ND5 gene was absent in the new mitogenome. These peculiar features of P. megacephalus mtDNA were further studied among related anuran species by PCR amplification. The new sequence data was used to assess the phylogenetic relationships of the three living amphibian orders using neighbor-joining, maximum likelihood, and Bayesian methods. In agreement with most morphological studies, phylogenetic analyses of a whole mitochondrial genome data set suggest a close relationship between salamanders and frogs. Moreover, using a molecular clock-independent Bayesian approach for inferring dating information from molecular phylogenies, we have provided a rough timescale for living amphibian evolution. This timescale provides a working framework for future paleontological researches on amphibian evolution and improves our understanding of the evolutionary history of modern amphibians.

Animals↗
PubMed2005

Mitochondrial phylogeny of Anura (Amphibia): a case study of congruent phylogenetic reconstruction using amino acid and nucleotide characters.

We explore whether phylogenetic analyses of the same sequence data set at the amino acid and nucleotide level are able to recover congruent topologies, as well as the advantages and limitations of both alternative approaches. As a case study, mitochondrial protein-coding genes were used to discern among competing hypotheses on the phylogenetic relationships of major anuran amphibian lineages. To properly address this phylogenetic question, the complete nucleotide sequences of the mitochondrial genomes of two archaeobatrachian species, Ascaphus truei and Pelobates cultripes, were determined anew. Bayesian and maximum likelihood phylogenetic inferences of the same sequence data set were performed based on both amino acid and nucleotide characters, with the latter analysed either as codons or as a reduced data set of first+second (P12) codon positions. In addition, likelihood-based ratio tests were performed to evaluate the support of alternative topologies. The different data sets arrived at congruent and highly supported topologies, suggesting a similar phylogenetic resolving power of the two character types provided that correctly selected sites and appropriate evolutionary models are used. The reconstructed anuran mitochondrial phylogeny supports the paraphyly of Archaeobatrachia, with Ascaphus as sister group to all the remaining anurans, and Pelobates as sister group of Neobatrachia. However, the employed tree reconstruction methods and likelihood-based ratio tests seemed to be negatively affected by the fast evolving sequences of neobatrachians, suggesting that the phylogeny of Anura here presented is not definitive, and needs further investigation using an extended taxon sampling.

Amphibian Proteins↗
PubMed2005

Structural aspects of an unusual choroidal polarized hyaline cartilage in the eyeball of Bufo ictericus and Rana catesbeiana (Amphibia, Anura): morphological and physiological significance.

Amphibian eyes play an important role in vision and in several physiological processes, such as food capture and breathing. To maintain the integrity of the eyeball there is a unique cuplike hyaline cartilage as a supporting tissue. In Bufo ictericus and Rana catesbeiana the cartilage layer is located between the retina and the choroids, being designated as choroidal cartilage, important to visual performance. On the retinal surface, there is no perichondrium, and the pigmented epithelium exhibits an intimate relationship to the cartilage layer.

Animals↗
PubMed2005

Morphology of Bufo ictericus integument (Amphibia, Bufonidae).

Bufo ictericus integument was investigated by stereoscopic, low vacuum scanning electron microscopy, transmission electron microscopy and light microscopy. The studies revealed, that the dorsal integument surface is rougher than ventral. Three types of projections are visualized: larger rounded verrucae, smaller conical cornified tubercles, and conical short spines. Prominent verrucae are observed on the dorsal surface, being flatter on the ventral surface. The tubercles are visualized only on the dorsal surface. The verrucae are separated by grooves that may contribute spreading and retention of the glandular secretion upon the integument. The pattern of the epidermal grooves is also important for water distribution, protecting the animal against desiccation. The epidermis is composed of a stratified epithelium with intraepithelial blood vessels, where keratinocytes predominate, but flask cells, and Merkel cells also occur. In the spongious dermis, cutaneous glands are visualized. The compact dermis is a series of alternating layers of bundles of collagenous fibers, and between spongious and compact dermis there are basophilic areas that correspond to Eberth-Katschenko layer. The dorsal and the ventral surfaces of B. ictericus are morphologically distinct. The integument structure is related to the physiology of each surface and represents an adaptation to habitat, reflecting a lifestyle of the animal.

Animals↗
PubMed2006

Morphological characterization of the anuran integument of the Proceratophrys and Odontophrynus genera (Amphibia, Anuran, Leptodactylidae).

The morphological characteristics of the leptodactylid integument of Proceratophrys and Odontophrynus genera were investigated by means of stereoscopic, low vacuum scanning electron and light microscopy. The integument surface of Proceratophrys boiei, Proceratophrys laticeps and Proceratophrys appendiculata exhibited several projections, while the integument of Odontophrynus americanus had rounded elevations with smooth profile. Light microscopic observations showed the basic integument morphology for all anurans, i.e., an epidermis and a dermis, which is subdivided into a spongious layer and a compact layer. The epidermis is formed by basal, intermediary and cornified layers. However, in Proceratophrys genus the cornified layer had an irregular outline, while in O. americanus the external surface was smooth. In the spongious dermis, mucous and venom exocrine glands were observed, but in O. americanus an exclusive glandular type with apocrine secretory pattern was identified. The integument morphology showed peculiar characteristics that may be helpful for genus distinction. Thus, morphological methods may be considered as an efficient means to characterize and to differentiate anuran genera.

Animals↗