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PubMed1990

Toad urinary bladder as a model for studying transepithelial sodium transport.

Sodium ion transport across tight epithelia has been investigated particularly extensively by studying two model systems: the urinary bladder of the toad and the frog skin. The greatest advantage presented by these models is the capability of monitoring net transepithelial Na+ flux simply, precisely, and instantaneously by measurement of the short circuit current (ISC). Many of the caveats involved in the measurement are discussed in detail. In order to fully characterize the forces driving Na+ movement across the series apical and basolateral membranes, it is necessary to measure intracellular potential and ionic composition. Such measurements are far more easily conducted with frog skin than with toad bladder, using the major biophysical techniques currently available. Regulation of transepithelial Na+ movement across tight epithelia is largely conducted at the apical membranes. This regulation can be clarified by study of the isolated Na+ channels in membrane vesicles. Such vesicles are far more easily prepared from toad urinary bladder than from frog skin. The strengths and potential misappropriations of this technique are considered in detail.

Animals↗
PubMedDate not supplied

[Oocyte maturation and activation in the common frog and the clawed toad under the action of divalent cations].

Maturation of Rana temporaria and Xenopus laevis oocytes was induced by solutions containing Mn2+ and Co2+ ions. Completion of oocyte maturation was estimated by the following criteria: (1) appearance of the maturation promoting factor (MPF) in the oocyte cytoplasm and (2) oocyte capacity to activation and formation of male pronuclei from the injected sperm nuclei. X. laevis oocytes matured under the effect of Co2+ ions were shown to contain MPF. Oocytes of both species matured under the effect of either ions could not be activated by pricking with a needle and injected sperm nuclei didn't transform into pronuclei. R. temporaria oocytes matured under the effect of ions in late spring, when natural spawning takes place, showed spontaneous activation.

Animals↗
PubMedDate not supplied

[The role of cAMP in suppressing the maturation of follicle-enclosed oocytes in the common frog and the clawed toad after their treatment with actinomycin D].

During maturation of the follicle-enclosed Rana temporaria and Xenopus laevis oocytes stimulated by pituitary suspension or human chorionic gonadotropin, respectively, the level of cAMP (follicle+medium) increased. When maturation was stimulated by low concentrations of gonadotropic hormones (inducing; however, maturation in 100% or close to 100% cases), the cAMP level increased 1.5-3 fold. Both the increased concentration of gonadotropic hormones and pretreatment of the follicles with actinomycin D stimulated further increase in cAMP level. These treatments in combination led to a significant increase of cAMP and inhibition of oocyte maturation.

Animals↗
PubMedDate not supplied

[The effect of a chloride channel inhibitor and of media with a decreased concentration of chlorine ions on spontaneous oocyte maturation in the common frog and the clawed toad].

Chloride-deficient media (irrespective of anions substituted for chloride) increased the rate of spontaneous maturation of follicle-enclosed (FEOs) and denuded oocytes in Rana temporaria and Xenopus laevis. The chloride channel blocker SITS (4-acetamido-4'-isothiocyanostilbene-2,2'-disulphonic acid) (10 microM) significantly decreased the rate of the spontaneous maturation of the FEOs and oocytes. The addition of 10 mM (instead of 2.7 mM) calcium ions to the gluconate solution significantly increased the rate of the spontaneous maturation of FEOs of R. temporaria if it was low in the gluconate solution, and decreased it if it was close to 100%.

4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfo↗
PubMed1998

Spatial distribution of yolk platelets and fat droplets in oocytes and cleaving embryos of the common frog (Rana temporaria) and toad (Bufo bufo).

Animal-vegetal gradients of fat droplets and yolk platelets have been quantitatively determined in mature oocytes and in cleaving embryos in two anuran species Rana temporaria and Bufo bufo using stereological methods. Volume densities Vv which describe properly the amount of nutrients in embryos have been used during the measurements. The embryos till the stage of late blastula are spherical: the spherical form is not distorted even by blastocoel forming eccentrically, in the animal hemisphere. Increased diameters during cleavage, which have been observed in the two species, can be ascribed to growth of the blastocoel and increased number of blastomeres while the quantity of the cytoplasm does not increase. The volume densities of yolk platelets and fat droplets per unit of cytoplasm volume remains constant throughout the cleavage. Volume densities change along the animal-vegetal axis in accordance with the course of the 3rd polynomial depending on the distance of the area under study from the vegetative pole f: y-->Vv, f = ay3 + by2 + cy + d. Parameters of fitting functions a, b, c and d change considerably during development of the embryo, which proves reorganisation of the cytoplasm during cleavage: the most significant changes occur from fertilisation to the four-cell stage and during growth of the blastocoel from morula to late blastula stage. Distortion of the axial symmetry of yolk platelet distribution in embryos occurring after fertilisation is not extensive in the two studied species. The mathematical model of spatial distribution of yolk platelets and fat droplets in oocytes and cleaving embryos in the studied anuran species was proposed using the above data.

Animals↗
PubMed1979

Light and scanning microscopy of the tongue and its gustatory organs in the common toad, Bufo bufo (L.).

Unlike frogs and European tree frogs, the common toad Bufo bufo possesses a tongue lacking filiform papillae on its dorsal surface. Instead, the mucosal epithelium forms irregular, high folds, dividing the surface of the tongue into numerous separate compartments. At the bottom of those compartments occur openings of tubular glands and singular ciliated cells. In a strongly distended tongue the folds of mucosa disappear, and the regions of glandular orifices assume a shape of pocket-like recesses. The taste discs with an average diameter of 120 micrograms are localized on the top of markedly shortened fungiform papillae. The superficial layer of taste discs contains a single type of glandular supporting cells, which in part of discs show features suggesting their gradual dezintegration, probably related to the process of cellular turnover in gustatory organs.

Animals↗
PubMed2001

Pesticides are involved with population declines of amphibians in the California Sierra Nevadas.

Several species of frogs and toads are in serious decline in the Sierra Nevada Mountains of California. These species include the threatened red-legged frog ( Rana aurora ), foothill yellow-legged frog ( R. boylii ), mountain yellow-legged frog ( R. muscosa ), Cascades frog ( Rana cascadae ), western toad ( Bufo boreas ) and Yosemite toad ( B. canorus ). For many of these species current distributions are down to 10% of historical ranges. Several factors including introduced predators, habitat loss, and ultraviolet radiation have been suggested as causes of these declines. Another probable cause is air-borne pesticides from the Central Valley of California. The Central Valley, especially the San Joaquin Valley, is a major agricultural region where millions of pounds of active ingredient pesticides are applied each year (http://www.cdpr.ca.gov/dprdatabase.htm). Prevailing westerly winds from the Pacific Coast transport these pesticides into the into the Sierras.

Animals↗
PubMed2002

Phylogenetic position of the giant anuran trypanosomes Trypanosoma chattoni, Trypanosoma fallisi, Trypanosoma mega, Trypanosoma neveulemairei, and Trypanosoma ranarum inferred from 18S rRNA gene sequences.

Phylogenetic relationships within the kinetoplastid flagellates were inferred from comparisons of small-subunit ribosomal RNA gene sequences. These included 5 new gene sequences, Trypanosoma fallisi (2,239 bp), Trypanosoma chattoni (2,180 bp), Trypanosoma mega (2,211 bp), Trypanosoma neveulemairei (2,197 bp), and Trypanosoma ranarum (2,203 bp). Trees produced using maximum-parsimony and distance-matrix methods (least-squares, neighbor-joining, and maximum-likelihood), supported by strong bootstrap and quartet-puzzle analyses, indicated that the trypanosomes are a monophyletic group that divides into 2 major lineages, the salivarian trypanosomes and the nonsalivarian trypanosomes. The nonsalivarian trypanosomes further divide into 2 lineages, 1 containing trypanosomes of birds, mammals, and reptiles and the other containing trypanosomes of fish, reptiles, and anurans. Among the giant trypanosomes, T. chattoni is clearly shown to be distantly related to all the other anuran trypanosome species. Trypanosoma mega is closely associated with T. fallisi and T. ranarum, whereas T. neveulemairei and Trypanosoma rotatorium are sister taxa. The branching order of the anuran trypanosomes suggests that some toad trypanosomes may have evolved by host switching from frogs to toads.

Animals↗
PubMed1992

Comparative effects of mast cell degranulators on perfused systemic blood vessels of Bufo melanostictus and Rana tigrina.

Three agents known to induce release of mast cell constituents, viz. polymyxin, compound 48/80 and polysorbate-80, were evaluated for effect on perfused blood vessels of R. tigrina and B. melanostictus. The mast cell degranulators caused vasoconstriction in frog and toad, except that for P-80 whose responses in toad were equivocal. Toads showed a general low responsiveness in comparison to frogs. Pharmacologic intervention with pheniramine, metergoline, hydergine, atropine and mecamylamine, respectively ruled out role of histamine, 5-HT, catecholamine or acetylcholine or even autonomic mechanisms in the above phenomena. The observations are suggestive of phylogenetic differences in biochemical profile of mast cells in amphibian species.

Animals↗
PubMed1985

Cell volume regulation in frog urinary bladder.

We have studied the problem of cell volume homeostasis in toad and frog urinary bladder by using electrophysiological measurements and an optical measure of cell volume. After osmotically induced swelling, urinary bladder cells spontaneously regulate their volume through a net loss of potassium, chloride, and water. During inhibition of sodium transport by amiloride the cells swell to the same extent as controls, but the volume-regulatory process is blocked. Electrophysiological results under isosmotic conditions indicate that basolateral membrane resistance increases simultaneously with the amiloride-induced rise in apical membrane resistance during transport inhibition. These independent observations indicate that inhibition of apical membrane sodium entry results in a secondary decrease in basolateral membrane potassium permeability. When cells are exposed to calcium-free, hyposmotic Ringer's solution, cell volume regulation is blocked; subsequent addition of the calcium ionophore A23187 is ineffective in restoring the regulatory process. The ionophore does induce volume regulation, however, in amiloride-inhibited, osmotically swollen cells in the presence of external calcium. Calcium thus seems to control basolateral membrane potassium permeability and may be the intracellular mediator of apical and basolateral membrane interactions.

Amiloride↗
PubMedDate not supplied

[Influenza in heterothermic animals].

The objective was to study Orthomyxovirus in heterothermic animals. Blood samples from snakes (genus Bothrops and Crotalus) and from toads and frogs (genus Bufo and Rana) were collected to evaluate the red cell receptors and antibodies specific to influenza virus by the hemagglutination and hemagglutination inhibition tests, respectively. Both snakes and toads kept in captivity presented receptors in their red cells and antibodies specific to either influenza virus type A (human and equine origin) or influenza type B. The same was observed with recently captured snakes. Concerning the influenza hemagglutination inhibition antibodies protective levels were observed in the reptiles' serum, against influenza type A and type B. Unlike the toads, 83.3% of the frogs presented mean levels of Ab 40HIU for some influenza strains. It was concluded that heterothermic animals could offer host conditions to the influenza virus and also susceptibility to the infection.

Animals↗
PubMed1997

Corneal curvatures and refractions of central American frogs.

We employed neutralizing infrared videophotorefraction and photokeratometry to examine the manifest refractions and corneal curvatures of 21 species of anurans (frogs and toads) in five families (Dendrobatidae, Bufonidae, Centrolenidae, Leptodactylidae, and Hylidae) resident in Central America. We found that all of the anurans exhibited hyperopic refractions in air, but that the observed hyperopia was not totally explained by the small eye artefact (Glickstein & Millodot, 1970). An allometric comparison of the corneal radii of these small anurans with those of a large number of other vertebrates, inferred from ocular axial lengths, showed that their corneal radii increased significantly more rapidly with increasing body size than that of other vertebrates generally (allometric slope constants: anurans: 0.270 +/- 0.032; other vertebrates: 0.151 +/- 0.004). Among the anurans examined, nocturnal Hylids had significantly larger eyes than diurnal Dendrobatid frogs and Bufonid toads.

Animals↗
PubMed1988

Role of vesicular transport in ADH-stimulated aggregate delivery.

It has been assumed from studies in toad bladder that antidiuretic hormone (ADH)-stimulated particle delivery to the luminal membrane is mediated by particle-carrying tubular structures (aggrephores). We report studies in frog and toad urinary bladder showing that vesicles, rather than aggrephores, appear to play the major role in particle delivery in the frog and that vesicle and aggrephore delivery proceed in parallel in the toad. Our principal evidence for this view is that in the frog, transmission electron microscopy shows virtually no fused aggrephores. Supporting evidence includes the following. 1) Freeze-fracture studies show that the diameters of fusion events delivering particles can be quite small, indicating that they are formed by fused vesicles rather than fused aggrephores. 2) A significant population of small fusion events is also seen in the toad, along with larger fusion events related to both aggrephores and large vesicles. 3) Surface aggregate areas in both species are small, consistent with vesicular delivery. 4) Freeze-fracture replicas indicate delivery from shallow pits. We propose a system of transport of particles in which aggrephores act largely as intermediate storage organelles in the frog and as storage and fusion organelles in the toad.

Animals↗
PubMed2002

Presence of thyrotropin-releasing-hormone-immunoreactive (TRHir) amacrine cells in the retina of anuran and urodele amphibians.

The presence of thyrotropin-releasing-hormone-immunoreactive (TRH-ir) amacrine cells in the retina of amphibians is reported for the first time. The anuran and urodele retinas studied exhibit major differences in the distribution of TRH-ir cells. In the two urodele species investigated, most TRH-ir amacrine cells were located in the ganglion cell layer (GCL). These pear-shaped cells originate a dense TRH-ir dendritic plexus in strata 4-5 of the inner plexiform layer (IPL). A small number of TRH-ir amacrine cells were observed in the inner nuclear layer (INL). Most of these INL TRH-ir cells were multipolar neurons with radiating dendrites that originate a loose plexus in the IPL stratum 1. In the three anuran species investigated, most TRH-ir amacrine cells were located in the INL. Distribution of TRH-ir processes in the IPL of anurans was not so clearly layered as in urodeles, dendrites being observed throughout strata 1-5. In the toad retina THR-ir material was also observed in the outer plexiform layer, which suggests that toads may have some TRH-ir interplexiform neurons. In the frog and toad, TRH-ir fibers were also observed in the optic nerve, although their origin could not be ascertained. The number of TRH-ir amacrine cells per whole retina was higher in anurans than in urodeles, though urodeles have higher cell densities. The marked differences in distribution of TRH-ir amacrine cells observed between anurans and urodeles, and among the three anuran species, suggest different functions of TRH in retinal processing, perhaps related to the different specializations of the visual systems of these species.

Amacrine Cells↗
PubMed1998

alpha1-adrenoceptors antagonize activated chloride conductance of amphibian skin epithelium.

The effects of adrenoceptor agonists on the transepithelial Cl- conductance (GCl) in the skin of several amphibian species, both toads and frogs, were studied. Epinephrine (Epi) from the serosal side selectively and reversibly inhibited the voltage-activated GCl in toad skin and the short-circuit GCl in frog skin. The main effects of activation of the adrenoceptors must reside in the skin epithelium and not in the glands, since measurements were made both from intact skins and split epithelia with essentially the same results. Effective concentrations of Epi were variable among individual tissues. GCl was reduced to 34+/-17% (n=46) with 1 micromol/l Epi, but in some tissues 0.1 micromol/l inhibited more than 80% of GCl, whereas some preparations were little influenced at >3 micromol/l Epi. The affected receptor type was identified by the use of the alpha1-agonist phenylephrine, which mimicked the response of Epi at concentrations above 30 micromol/l, whereas the alpha2-agonists xylazine and iodoclonidine had no effect at supramaximal concentrations. Prazosin, a specific alpha1-antagonist, reduced or eliminated the inhibition by Epi, but the response pattern suggests a low affinity. The alpha2-antagonist yohimbine, at concentrations </=0.3 micromol/l, had a minimal effect, but reduced the inhibition by Epi at concentrations of 1-10 micromol/l. This might indicate affinity to alpha1-adrenoceptors in amphibian skin. Activation of beta-adrenoceptors by isoproterenol (0.1-5 micromol/l) led to a transient increase of the baseline inactivated component of GCl with a slight reduction of the voltage-activated GCl at the higher concentrations, but the inhibitory effect of Epi was not altered. Epi, on the other hand, neither prevented nor reversed the induction of a voltage-insensitive GCl in toad skin caused by application of cAMP at supramaximal concentrations (>100 micromol/l CPT-cAMP). Preincubation of the serosal medium with Ca2+-free solution (in the presence of 2 mmol/l EGTA) for extended periods of time (>30 min) eliminated the response to Epi. It is concluded that alpha1-adrenoceptors participate in the physiological control of voltage-activated Cl- conductance in amphibian skin epithelium via modulation of intracellular Ca2+, presumably by efflux from intracellular stores.

1-Methyl-3-isobutylxanthine↗
PubMed2003

Helminth community structure of sympatric eastern American toad, Bufo americanus americanus, northern leopard frog, Rana pipiens, and blue-spotted salamander, Ambystoma laterale, from southeastern Wisconsin.

One-hundred twelve amphibians, including 51 blue-spotted salamanders, Ambystoma laterale, 30 eastern American toads, Bufo americanus americanus, and 31 northern leopard frogs, Rana pipiens, were collected during April-October 1996 from Waukesha County, Wisconsin and examined for helminth parasites. The helminth compound community of this amphibian assemblage consisted of at least 10 species: 9 in American toads, 8 in leopard frogs, and 3 in blue-spotted salamanders. American toads shared 7 species with leopard frogs, and 2 species occurred in all 3 host species. Although there was a high degree of helminth species overlap among these sympatric amphibians, statistically significant differences were found among host species and percent of indirect or direct-life cycle parasites of amphibian species individual component communities (chi2 = 1,015, P < 0.001). American toads had a higher relative abundance of nematodes, 59%, than larval cestodes, 31%, and larval and adult trematodes, 10%, whereas leopard frogs had a higher relative abundance of larval cestodes, 71.3%, and larval and adult trematodes, 25.3%, than nematodes 3.4%. This is related to ecological differences in habitat and dietary preferences between these 2 anuran species. Helminth communities of blue-spotted salamanders were depauperate and were dominated by larval trematodes, 94%, and few nematodes, 6%. Low helminth species richness in this host species is related to this salamander's relatively small host body size, smaller gape size, lower vagility, and more fossorial habitat preference than the other 2 anuran species. Adult leopard frogs and toads had significantly higher mean helminth species richness than metamorphs, but there was no significant difference in mean helminth species richness among adult and metamorph blue-spotted salamanders. Considering adult helminths, the low species richness and low vagility of caudatans as compared with anurans suggest that local factors may be more important in structuring caudatan helminth communities of salamanders than of anuran hosts. Helminth species infecting salamanders may be more clumped in their geographic distribution as compared with anurans, and the role of other hosts and their parasites at the compound community level may be important in structuring helminth communities of salamanders.

Ambystoma↗
PubMed1979

[Injection of mitochondria into oocytes and fertilized eggs].

The suspension of mitochondria isolated from the loach embryos or the frog heart were injected in the oocytes or fertilized eggs of the loach, newt, toad and frog in the amount roughly equivalent to the content of mitochondria in the egg. After the injection the oocytes did not differ during several days from the normal ones and the fertilized eggs of the loach, newt and South Afican clawed toad developed normally. The activity of cytochrome oxidase in the injected oocytes was kept at a somewhat higher level (1.4 to 1.9 vs 1.0 in the control) during several days. In the developing eggs the activity of cytochrome oxidase began to decrease from the blastula stage and attained rapidly the control level. The decrease of the enzyme activity is due to non-specific degradation of excessive mitochondria or to compensatory inactivation of the enzyme ensuring the maintenance of its normal activity during the development.

Animals↗