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PubMed1984

Ectodermal fragments from normal frog gastrulae condition substrata to support normal and hybrid mesodermal cell migration in vitro.

Using time-lapse cinemicrography and scanning electron microscopy, we have shown that normal Rana embryos and gastrulating hybrid embryos have extracellular fibrils on the inner surface of the ectodermal layer. These fibrils are absent prior to gastrulation and appear in increasing numbers during gastrulation. They can also be deposited in vitro where they condition substrata in such a way that normal presumptive mesodermal cells placed on them show extensive attachment and unoriented cell movement. These fibrils are also present in some arrested hybrid embryos, but in reduced numbers, or are lacking in other arrested hybrid embryos. Explanted ectodermal fragments from arrested hybrid embryos fail both to condition culture substrata by the deposition of fibrils and to promote cell attachment and translocation. In contrast, ectodermal fragments from normal embryos can condition culture substrata so as to promote moderate cell attachment and, for one particular gamete combination, even cell translocation of presumptive mesodermal cells taken from arrested hybrid embryos. These results provide new evidence to support the hypothesis that extracellular fibrils represent a system that promotes mesodermal cell migration in amphibian embryos. Differences in the fibrillar system in urodele and anuran embryos are discussed in relation to fundamental differences in the mode of mesodermal cell migration in these two classes of Amphibia.

Animals↗

PubMed2004

Dogmas and controversies in the handling of nitrogenous wastes: expression of arginase Type I and II genes in rainbow trout: influence of fasting on liver enzyme activity and mRNA levels in juveniles.

Through analysis of a cDNA library and third-party annotation of available database sequences, we characterized the full-length coding regions of rainbow trout (Oncorhynchus mykiss) Type I, Onmy-ARG01, and Type II, Onmy-ARG02, arginase genes. Two partial related arginase sequences, Onmy-ARG01b and Onmy-ARG02b, and a full-length zebrafish arginase coding region (Danio rerio), Dare-ARG02, are also reported. Comparison of vertebrate arginase sequences shows that both Type I and Type II genes in bony fishes contain a mitochondrial targeting N-terminal domain. This suggests that the cytosolic Type I arginase found in ureotelic vertebrates arose in the common ancestor of amphibia and mammals. Onmy-ARG01 and Onmy-ARG02 mRNA was detected in liver, kidney, gill, intestine, red muscle and heart tissues. Onmy-ARG01 was expressed at a significantly higher level relative to Onmy-ARG02 in liver and red muscle tissue. To investigate whether there was differential regulation of Onmy-ARG01 and Onmy-ARG02, juvenile trout were fasted for 6 weeks and hepatic enzyme activities and mRNA levels were compared with those of fed control fish. There was a 3-fold increase in liver arginase activity and a 2-fold increase in Onmy-ARG02 mRNA levels but no change in Onmy-ARG01 mRNA levels in fasted fish relative to fed fish. These findings indicate that both types of arginase genes are present and expressed in rainbow trout and that the pattern of expression varies between tissues. The increase in liver arginase activity after a 6-week fast is due, in part, to an increase in the expression of Onmy-ARG02 mRNA levels.

Amino Acid Sequence↗

PubMed1984

Functional conflicts between feeding and gas exchange in suspension-feeding tadpoles, Xenopus laevis.

Air-breathing tadpoles of Xenopus laevis (Amphibia: Anura) use buccopharyngeal surfaces for both gas exchange and capture of food particles in the water. In dense food suspensions, tadpoles decrease ventilation of the buccopharynx and increase air breathing. The lung ventilatory frequency is elevated even though the rate of oxygen consumption is at or below resting levels, suggesting that the lung hyperventilation reflects compensation for decreased buccopharyngeal respiration rather than an increased metabolic requirement. If tadpoles in hypoxic water are prevented from breathing air, they increase buccopharyngeal respiration at the expense of feeding. Aerial respiration evidently permits the buccopharyngeal surfaces to be used primarily for food entrapment.

Animals↗

PubMed2001

Characteristic features and ligand specificity of the two olfactory receptor classes from Xenopus laevis.

Amphibia have two classes of olfactory receptors (ORs), class I (fish-like receptors) and class II (mammalian-like receptors). These two receptor classes correspond to the two classes identified in other vertebrates, and amphibians thus provide a unique opportunity to compare olfactory receptors of both classes in one animal species, without the constraints of evolutionary distance between different vertebrate orders, such as fish and mammals. We therefore identified the complete open reading frames of class I and class II ORs in Xenopus laevis. In addition to allowing a representative comparison of the deduced amino acid sequences between both receptor classes, we were also able to perform differential functional analysis. These studies revealed distinct class-specific motifs, particularly in the extracellular loops 2 and 3, which might be of importance for the interaction with odorants, as well as in the intracellular loops 2 and 3, which might be responsible for interactions with specific G-proteins. The results of functional expression studies in Xenopus oocytes, comparing distinct receptor types, support the idea that class I receptors are activated by water-soluble odorants, whereas class II receptors are activated by volatile compounds.

Amino Acid Sequence↗

PubMed1994

Eutherian phylogeny as inferred from mitochondrial DNA sequence data.

The phylogenetic relationships among Primates, Artiodactyla, Cetacea, Carnivora, and Rodentia were estimated from the amino acid sequences of proteins encoded by the mitochondrial genomes, for which entire nucleotide sequence data are available, using Marsupialia, Aves, and Amphibia as outgroups. The overall evidence of the maximum likelihood, as well as maximum parsimony, analyses strongly suggests that Rodentia is an outgroup to the other four eutherian orders, and that Cetacea and Artiodactyla form a clade with Carnivora as a sister taxon, consistently with the molecular phylogenetic studies of previous authors. However, analyses of individual proteins do not necessarily conform to this conclusion, and some of the proteins reject the putatively correct tree with nearly 5% significance. Furthermore, the 12S mitochondrial ribosomal RNA sequences do not give the putatively correct tree irrespective of the alignments and of the phylogenetic methods, although the tree is not rejected with a statistical significance. The 16S ribosomal RNA sequences give the putatively correct tree with our sequence alignment when the ML method is used, but the result depends on the alignment and on the choice of outgroup species. These results illustrate the limitation of the ribosomal RNA data alone in phylogenetic inference, and suggest that we must analyze as many genes as possible and synthesize the results to draw a reliable conclusion.

Amino Acid Sequence↗

PubMed2000

Ecological risk assessment of endocrine disruptors.

The European Centre for Ecotoxicology and Toxicology of Chemicals proposes a tiered approach for the ecological risk assessment of endocrine disruptors, integrating exposure and hazard (effects) characterization. Exposure assessment for endocrine disruptors should direct specific tests for wildlife species, placing hazard data into a risk assessment context. Supplementing the suite of mammalian screens now under Organization for Economic Cooperation and Development (OECD) validation, high priority should be given to developing a fish screening assay for detecting endocrine activity in oviparous species. Taking into account both exposure characterization and alerts from endocrine screening, higher tier tests are also a priority for defining adverse effects. We propose that in vivo mammalian and fish assays provide a comprehensive screening battery for diverse hormonal functions (including androgen, estrogen, and thyroid hormone), whereas Amphibia should be considered at higher tiers if there are exposure concerns. Higher tier endocrine-disruptor testing should include fish development and fish reproduction tests, whereas a full life-cycle test could be subsequently used to refine aquatic risk assessments when necessary. For avian risk assessment, the new OECD Japanese quail reproduction test guideline provides a valuable basis for developing a test to detecting endocrine-mediated reproductive effects; this species could be used, where necessary, for an avian life-cycle test. For aquatic and terrestrial invertebrates, data from existing developmental and reproductive tests remain of high value for ecological risk assessment. High priority should be given to research into comparative endocrine physiology of invertebrates to support data extrapolation to this diverse fauna.

Amphibians↗

PubMed1987

Teratological research using in vitro systems. V. Nonmammalian model systems.

In this review of alternative tests to whole-animal rodent studies, the use of sub-mammalian and sub-vertebrate systems is investigated. The history, methodology, known limitations, end points, dose response, and requirements of virus, hydra, planarian, cricket, fish, amphibia, Drosophila, and chicken embryo systems are discussed.

Amphibians↗

PubMed2006

Nuclear transplantation in Xenopus.

Nuclear transplantation in amphibia started in 1952. By this is meant, sexually mature cloned frogs can be obtained from the nuclei of embryo cells, differentiating cells, and larval-differentiated cells. Transplanted nuclei are reprogrammed to entirely new patterns of gene expression. In this chapter, the methods used to transplant living nuclei into enucleated eggs of Xenopus are described. A method also is described for transplanting multiple somatic cell nuclei into nonenucleated oocytes, a procedure that achieves reprogramming of gene expression in the absence of cell division.

Active Transport, Cell Nucleus↗

PubMed2000

Evolution of urate-degrading enzymes in animal peroxisomes.

The end product of purine metabolism varies from species to species. The degradation of purines to urate is common to all animal species, but the degradation of urate is much less complete in higher animals. The comparison of subcellular distribution, intraperoxisomal localization forms, molecular structures, and some other properties of urate-degrading enzymes (urate oxidase, allantoinase, and allantoicase) among animals is described. Liver urate oxidase (uricase) is located in the peroxisomes in all animals with urate oxidase. On the basis of the comparison of intraperoxisomal localization forms, mol wt, and solubility of liver urate oxidase among animals, it is suggested that amphibian urate oxidase is a transition form in the evolution of aquatic animals to land animals. Allantoinase and allantoicase are different proteins in fish liver, but the two enzymes form a complex in amphibian liver. The subcellular localization of allantoinase and allantoicase varies among fishes. Hepatic allantoinase is located both in the peroxisomes and in the cytosol in saltwater fishes, and only in the cytosol in freshwater fishes. Hepatic allantoicase is located on the outer surface of the peroxisomal membrane in the mackerel group and in the peroxisomal matrix in the sardine group. Amphibian hepatic allantoinase-allantoicase complex is probably located in the mitochondria. On the basis of previous data, changes of allantoinase and allantoicase in molecular structure and intracellular localization during animal evolution may be as follows: Fish liver allantoinase is a single peptide with a mol wt of 54,000, and is located both in the peroxisomes and in the cytosol, or only in the cytosol. Fish liver allantoicase consists of two identical subunits with a mol wt of 48,000, and is located in the peroxisomal matrix or on the outer surface of the peroxisomal membrane. The evolution of fishes to amphibia resulted in the dissociation of allantoicase into subunits, and in the association of allantoinase with the subunit of allantoicase. This amphibian enzyme was lost by further evolution.

Amidohydrolases↗

PubMed1978

[Effect of caerulein on pancreatic endocrine and exocrine secretion from the perfused rat pancreas (author's transl)].

Several investigations in vivo and in vitro have shown that gastro-intestinal hormones stimulate insulin secretion. However, the reports on the insulinotropic activity of pancreozymin are contradictory. The conflicting results are probably due to the fact that pure native preparation of this hormone has not been obtained in "physiologic" doses. In the present study this problem has been investigated by exposing rat pancreas to caerulein in vitro. Caerulein, an active decapeptide isolated from the skin of the Australian amphibia Hyla caerulea, resembles pancreozymin in chemical structure, including C-terminus. This active polypeptide of nonmammalian origin has been shown to possess all the biological activities of pancreozymin. The present investigation was undertaken to evaluate the significance of the interactions of exocrine and endocrine pancreas using perfused rat pancreas in vitro. Biphasic insulin release was demonstrated with caerulein at concentrations higher than 1 ng/ml. Insulin response of the first phase was proportional to the dose up to 1 microgram/ml. The second phase of insulin release was, however, almost constant, regardless of the concentrations of caerulein. Release of glucagon was stimulated by the same concentrations of caerulein which stimulated insulin release. Maximal response of the pancreatic amylase and pancretic juice output were observed with 1 ng/ml of caerulein. With higher doses, significantly less secretory responses were observed. The dissociation of the response to caerulein between endocrine and exocrine pancreas was found.

Amylases↗

PubMed1996

Neoteny and the thyroid in ratites.

The ratites (for example ostriches, emus) are neotenous descendants of flying birds. The best studied cases of neoteny among vertebrates are in the Amphibia. In this class, whether individuals metamorphose and breed as adults, or whether they become sexually mature as neotenous aquatic larvae, is controlled by the thyroid. In this review it is argued that the thyroid may have been important in the evolution of ratite neoteny. The evidence is based on characteristics of ratites that could indicate thyroid abnormality, similarities between ratites and thyroidectomized non-ratite birds, and preliminary results from a study of thyroid function in ostriches.

Animals↗

PubMed2005

Priority areas for anuran conservation using biogeographical data: a comparison of greedy, rarity, and simulated annealing algorithms to define reserve networks in cerrado.

Spatial patterns in biodiversity variation at a regional scale are rarely taken into account when a natural reserve is to be established, despite many available methods for determining them. In this paper, we used dimensions of occurrence of 105 species of Anura (Amphibia) in the cerrado region of central Brazil to create a regional system of potential areas that preserves all regional diversity, using three different algorithms to establish reserve networks: "greedy", rarity, and simulated annealing algorithms. These generated networks based on complementarity with 10, 12, and 8 regions, respectively, widely distributed in the biome, and encompassing various Brazilian states. Although the purpose of these algorithms is to find a small number of regions for which all species are represented at least once, the results showed that 67.6%, 76.2%, and 69.5% of the species were represented in two or more regions in the three networks. Simulated annealing produced the smallest network, but it left out three species (one endemic). On the other hand, while the greedy algorithm produce a smaller solution, the rarity-based algorithm ensured that more species were represented more than once, which can be advantageous because it takes into consideration the high levels of habitat loss in the cerrado. Although usually coarse, these macro-scale approaches can provide overall guidelines for conservation and are useful in determining the focus for more local and effective conservation efforts, which is especially important when dealing with a taxonomic group such as anurans, for which quick and drastic population declines have been reported throughout the world.

Algorithms↗

PubMed1995

The phylogeny of alpha-fetoprotein in vertebrates: survey of biochemical and physiological data.

The phylogeny of vertebrate alpha-fetoprotein (AFP) was surveyed in the phylum Chordata, including subphyla, agnatha, and Gnathostoma. A molecular taxonomic approach was undertaken based on biochemical, endocrinological, immunological, and physiological criteria previously documented for AFP. These published data were then discussed in light of their position and relationship in the albuminoid gene superfamily derived from GenBank. The phylogeny of the AFP molecule should prove useful for investigators seeking markers for animal models of human diseases, serological cross-reactivity between AFP molecular species, identification of larval or fetal protein homologs of AFP, and provide strategies for biochemical purification and physiological studies. The phylogeny of AFP in vertebrates was surveyed from the cyclostomata (lamphrey) to the mammals, including sharks, bony fishes, amphibians, reptiles, and birds. A trend was denoted, from lower to higher animal forms, in a size reduction and separation of AFP-like albumin molecular moieties from forms that resembled true albumin molecules. While the primitive lamphrey possesses a serum protein twice the size of mammalian albumin, the bony fishes, reptiles, and amphibians display two ALB-like molecules sharing amino acid sequence similarity to mammalian AFP. However, only one of the ALB-like molecules in the fish and amphibia is glycosylated. Although little has been published on reptilian AFP-like molecules, avian AFP has been investigated extensively following its detection and isolation for developmental studies involving immunology and neuroendocrinology. Finally, a plethora of knowledge exists in mammals following several decades of studies involving the isolation, purification, and characterization of AFP for use in physiological, immunological, and endocrinological research endeavors. In overview, a tendency or trend in down-sizing of an AFP-like albumin molecule and the separation of true albumin forms from a distinct fetal glycosylated form was observed. This seemed to occur in animal classes lacking a free-swimming larval form and possessing either highly differentiated extra-embryonic membranes or displaying a placenta intimately interfaced with the maternal tissues of the uterus.

Animals↗

PubMed1984

Effects of synthetic mammalian thyrotrophin releasing hormone, somatostatin and dopamine on the secretion of prolactin and growth hormone from amphibian and reptilian pituitary glands incubated in vitro.

Pituitary glands of grassfrog (Rana pipiens), bullfrog (Rana catesbeiana), clawed toad (Xenopus laevis) and two species of terrapin (Chrysemys picta and Pseudemys scripta) were incubated in medium containing hypothalamic extract (HE), thyrotrophin releasing hormone (TRH), somatostatin, dopamine, or combinations of these treatments. Prolactin and GH concentrations in the medium were determined by densitometry after polyacrylamide-gel electrophoretic separation. Hypothalamic extract stimulated secretion of both hormones in all species tested. Thyrotrophin releasing hormone stimulated secretion of prolactin and GH, showing a biphasic pattern of response. Dopamine had little effect alone, but inhibited HE- and TRH-stimulated release of prolactin, but not GH, in both amphibia and reptiles. Somatostatin by itself had no apparent effect on release of hormones, but it inhibited HE- and TRH-stimulated release of GH from both amphibian and reptilian pituitary glands. These results indicate that factors affecting mammals and birds also interact in the regulation of secretion of prolactin and GH in lower vertebrate species.

Animals↗

PubMed1989

Chemoreceptive and mechanoreceptive paraneurons in the tongue.

Chemoreceptive and mechanoreceptive paraneurons in the tongue include gustatory cells and basal cells of the taste buds, Merkel cells in the lingual epithelium and Merkel or Grandry corpuscles in avian subepithelial connective tissue. Neuron-specific enolase immunoreactivity is recognized in the basal cells, a type of Merkel cells, in the taste buds of teleost fishes and amphibia, certain taste bud cells of mammals and the Merkel cells in birds. The present study also concerns itself with the ultrastructural features of the gustatory cells of the rabbit taste buds, and the Merkel cells of the finch tongue. The functional significance of the dense-cored granules in the gustatory cell is still enigmatic. On the other hand, the dense-cored granules in the Merkel cells may have dual functions storing both neurotransmitters and local hormones. Shortly after giving a mechanical stimulus to the finch tongue, we could demonstrate massive exocytotic changes of the Merkel cell granules.

Amphibians↗

PubMed1980

Ultrastructural and histochemical studies on the taste buds in some reptiles.

The taste buds in tortoises (Clemmys japonica and Geoclemys reevesii), lizards (Takydromus tachydromoides) and snakes (Elaphe quadrivirgata) were examined by both ultrastructural and histochemical methods. The taste buds consisted of at least three types of cells: the type I, II and III cells. The type I cells were characterized by the presence of secretory dense granules containing polysaccharides which were demonstrated by periodic acid-chromic acid-silver methenamine technique. The type II cells contained numerous tubular, vesicular and lamellated structures. The type III cells were characterized by dense cored vesicles and afferent synaptic contacts. Besides these cells, basally located cells which resembled the basal cells of other lower vertebrates were sometimes found in the tortoises. After administration of L-DOPA following nialamide, some taste bud cells of the tortoises, Clemmys japonica, showed weak yellowish green fluorescence by monoamine fluorescence histochemistry and the dense-cored vesicles in the type III cells increased in number. Acetylcholine esterase activity was not observed in tortoise taste buds. It is suggested that the three types of cells which compose the taste buds of the reptiles may correspond to the three types of cells in mammalian, and the type III cells represent the gustatory cells which are able to potentially produce biogenic monoamines. From these results, the taste buds of the reptiles may hold an intermediate position between those of mammals and amphibia or fishes.

5-Hydroxytryptophan↗

PubMed1983

The transmural potential of the newt ureter: evidence for amiloride-sensitive active sodium transport.

Transmural electrical potential difference (PD) of the newt ureter was measured with glass microelectrodes and its nature was studied pharmacologically or by ion replacement experiments. The PD in the presence of standard Ringer solution on both sides averaged -76.0 +/- 1.3 mV, lumen negative. The magnitude of the PD depended on the Na+ concentration of the luminal fluid; the PD increased in a saturable way with increase in the Na+ concentration. Amiloride added to the luminal fluid at 10(-4) M greatly reduced the PD and increased the transmural resistance. The half maximum inhibition was seen at 2.6 x 10(-6) M and the maximum inhibition at 10(-4) M where the PD reduced to 3.8% of control. Amiloride added to the external medium at 10(-4) M had no effect on both the PD and the resistance. Ouabain added to the external medium at 10(-3) M caused a gradual decrease in the PD, which fell to 36% of its initial value after 60 min. The specific transmural resistance (Rt) was estimated by recording spatially decaying electrotonic potentials along the ureter and applying cable analysis. The short-circuit current (Isc) was calculated from the PD and the Rt. The Rt and the Isc averaged 31.7 +/- 7.7 k omega X cm2 and 3.9 +/- 1.5 microA X cm-2, respectively. The results indicate that the epithelial cells of the newt ureter have amiloride-sensitive Na+ channels at the luminal membrane and are transporting Na+ actively, like epithelia of the urinary bladder and the skin of amphibia.

Amiloride↗

PubMed2002

PACAP and its receptors exert pleiotropic effects in the nervous system by activating multiple signaling pathways.

Pituitary adenylate cyclase-activating polypeptide (PACAP) was originally isolated from the ovine brain in 1989 as a novel hypothalamic hormone that potently activates adenylate cyclase to produce cyclic AMP in pituitary cells. This neuropeptide belongs to the secretin/glucagon/vasoactive intestinal peptide (VIP) superfamily, and exists in two amidated forms as PACAP38 (38-amino acid residues) and PACAP27 derived from the same precursor. The primary structure of PACAP has been remarkably conserved throughout evolution among tunicata, ichthyopsida, amphibia and mammalia, and a PACAP-like neuropeptide has also been determined in Drosophila. Both PACAP and its receptors are mainly distributed in the nervous and endocrine systems showing pleiotropic functions with high potency. There are three types of receptors with high PACAP-binding affinity and with different tissue-distribution patterns. All of them belong to G-protein-coupled receptor superfamily with seven transmembrane domains. PAC(1) is the PACAP-specific receptor and exists in at least eight splice variants which couple to different intracellular signal transduction pathways. VPAC(1) and VPAC(2) are the common receptors for both PACAP and VIP, which are coupled to adenylate cyclase. This review article presents and discusses an update on PACAP research and its pleiotropic physiological functions based on multiple receptor-mediated signaling mechanisms in both the central and peripheral nervous system, including the regulation of hypothalamic neurosecretion, homeostatic control of circadian clock and behavioral actions, involvement in learning and memory processes, neuroprotective effects such as anti-apoptosis and response to injury and inflammation, and neural ontogenetic functions on proliferation/differentiation processes from early stages.

Animals↗