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At least 127 records · Page 7Linked to original sources
PubMed1978

Membrane structural specialization of the toad urinary bladder revealed by the freeze-fracture technique. III. Location, structure and vasopressin dependence of intramembrane particle arrays.

Examination of the toad urinary bladder by freeze-fracture electron microscopy reveals intramembrane particle arrays at a number of membrane sites. An array in which particles are aggregated into closely apposed parallel rows is found in the granular cell luminal membrane of dehydrated toads fixed in situ. These aggregates are structurally indistinguishable from those previously associated with vasopressin exposure in vitro. Aggregates are not found in granular cell luminal membrane in the case of hydrated toads fixed in situ. However, structurally similar arrays are found at low frequency in the membrane of cytoplasmic vacuoles in granular cells and in the plasma membrane of basal cells in both hydrated and dehydrated toads. Aggregates are also present at these sites in control and vasopressin-treated bladders from in vitro experiments. Particle arrays characteristic of gap junctions, desmosomes and hemidesmosomes also occur in the plasma membrane of basal cells. In addition, distinctive square arrays of particles exist in the plasma membrane of the bladder's mesothelium. Although a variety of intramembrane particle arrays exist in the toad urinary bladder, only the occurrence of organized particle aggregates in the luminal membrane of granular cells appears to be associated with vasopressin exposure.

Animals↗
PubMed1998

Effect of nifedipine on depolarization-induced force responses in skinned skeletal muscle fibres of rat and toad.

The effect of the dihydropyridine, nifedipine, on excitation-contraction coupling was compared in toad and rat skeletal muscle, using the mechanically skinned fibre technique, in order to understand better the apparently disparate results of previous studies and to examine recent proposals on the importance of certain intracellular factors in determining the efficacy of dihydropyridines. In twitch fibres from the iliofibularis muscle of the toad, 10 microM nifedipine completely inhibited depolarization-induced force responses within 30 s, without interfering with direct activation of the Ca(2+)-release channels by caffeine application or reduction of myoplasmic [Mg2+]. At low concentrations of nifedipine, inhibition was considerably augmented by repeated depolarizations, with half-maximal inhibition occurring at < 0.1 microM nifedipine. In contrast, in rat extensor digitorum longus (EDL) fibres 1 microM nifedipine had virtually no effect on depolarization-induced force responses, and 10 microM nifedipine caused only approximately 25% reduction in the responses, even upon repeated depolarizations. In rat fibres, 10 microM nifedipine shifted the steady-state force inactivation curve to more negative potentials by < 11 mV, whereas in toad fibres the potent inhibitory effect of nifedipine indicated a much larger shift. The inhibitory effect of nifedipine in rat fibres was little, if at all, increased by the absence of Ca2+ in the transverse tubular (t-) system, provided that the Ca2+ was replaced with sufficient Mg2+. The presence of the reducing agents dithiothreitol (10 mM) or glutathione (10 mM) in the solution bathing a toad skinned fibre did not reduce the inhibitory effect of nifedipine, suggesting that the potency of nifedipine in toad skinned fibres was not due to the washout of intracellular reducing agents. The results are considered in terms of a model that can account for the markedly different effects of nifedipine on the two putative functions of the dihydropyridine receptor, as both t-system calcium channel and a voltage-sensor controlling Ca2+ release.

Animals↗
PubMed1999

Pharmacological and biochemical investigation of receptors for the toad gut tachykinin peptide, bufokinin, in its species of origin.

This is the first report of the development of a new radioligand [125I]Bolton-Hunter bufokinin ([125I]BH-bufokinin) and its use in the characterisation of tachykinin receptors in the small intestine of the cane toad, Bufo matrinus. The binding of [125I]BH-bufokinin to toad intestinal membranes was rapid, saturable, of high affinity and to a single population of binding sites with KD 0.57 nM and Bmax 3.1 fmol mg wet weight tissue(-1). The rank order of affinity of tachykinins to compete for [125I]-BH bufokinin binding revealed similarities with that of the mammalian NK1 receptor, being bufokinin (IC50, 1.7 nM)>physalaemin (6.7 nM)>substance P (SP, 10.7 nM)> or =neuropeptide gamma (NPgamma, 12.4 nM)> or =kassinin (17.8 nM)>scyliorhinin I (35.3 nM)> or =eledoisin (40.6 nM)> or =carassin (43.2 nM)> or =neurokinin A (NKA, 57.8 nM)> or =neurokinin B (NKB, 77.5 nM)>scyliorhinin II (338 nM). The mammalian NK3-selective agonist senktide was a very weak competitor. The radioligand [125I]neurokinin A showed no specific binding to toad intestinal membranes. In the toad isolated small intestine, the maximum contractile response to bufokinin was over 150% greater than that to acetylcholine in longitudinal muscle, whereas responses to bufokinin and acetylcholine were similar in circular muscle. Bufokinin was the most potent agonist (EC501 0.34 nM) and produced a long-lasting contraction. Other tachykinins such as physalaemin, SP and kassinin were also potent contractile agents. The potency values of mammalian and amphibian tachykinins derived from functional studies (pD2) correlated significantly with those from binding assays (pKi). The data for fish and molluscan tachykinins, however, showed poor correlation. Contractions to bufokinin and SP were unaffected by atropine, indomethacin and tetrodotoxin. The highly selective NK1 receptor antagonists CP 99994, GR 82334 and RP 67580 were ineffective in both binding and functional studies. Bufokinin increased inositol monophosphate formation in a concentration-dependent manner with an EC50 value of 10.7 nM, suggesting that the tachykinin receptor may be coupled to phosphoinositol hydrolysis. In summary, this study provides evidence for a high-affinity, bufokinin-preferring, NK1-like tachykinin receptor in the toad small intestine. This is probably not the receptor which mediates contraction to carassin, scyliorhinin II and eledoisin. The study also provides evidence that bufokinin and its receptor play an important physiological role in regulating intestinal motility.

Animals↗
PubMed2001

Lymph osmolality and rehydration from NaCl solutions by toads, Bufo marinus.

Toads, Bufo marinus, allowed to maintain an ad libitum state of hydration were dehydrated by 10 15% of their standard weight and allowed to rehydrate from either deionized water or from 10 or 50 mmol l(-1) NaCl solutions. Toads rehydrating from the dilute salt solutions recovered a larger fraction of their standard weight than did toads rehydrating from deionized water despite there being a reduced osmotic gradient. Amiloride did not reduce water gain from these solutions. Water uptake from 100 mmol l(-1) sucrose and 50 mmol l(-1) Na gluconate was reduced relative to deionized water by a fraction predicted from the osmotic gradient. Thus, the presence of both Na+ and Cl- are required for the augmentation of water gain from dilute salt solutions. Toads allowed to rehydrate from 120 mmol l(-1) NaCl for 180 min recovered nearly as much water as toads rehydrating from deionized water for 120 min and the lymph osmolality was not reduced relative to the dehydrated condition. The recovery of water from the salt solution was greater than that predicted from the reduced osmotic gradient and amiloride partially inhibited the rehydration from 120 mmol l(-1) NaCl. Solute coupled water transport can therefore be demonstrated in living animals but only from a NaCl solution that is nearly isoosmotic with the lymph. The mechanism for enhanced water gain from dilute salt solutions remains unresolved.

Amiloride↗
PubMed1980

The effects of bicarbonate and hydroxyl ions on chloride transport by toad bladders.

The association between Cl-, HCO3- and H+ transported by toad bladders was investigated. Net mucosal to serosal Cl- transport by Colombian toad bladders was stimulated by incubation in HCO3- free solutions. In addition, when Colombian or Dominican toad bladders were exposed to low HCO3- concentrations on the mucosal side and 25 mM HCO3- on the serosal side, net mucosal leads to serosal Cl- transport was induced. Neither acetazolamide nor cyanide significantly inhibited Cl- transport under these conditions. The presence of a pH gradient, more acid on the mucosal side, also induced net mucosal leads to serosal Cl- transport. The results suggest that Cl- transport by toad bladders may occur by exchange with HCO3- or OH-; this process may not require carbonic anhydrase or oxidative metabolism. The Cl- transport by toad bladders is qualitatively different from the electrogenic Cl- transport of the thick limb of Henle's loop, but may be similar to a process which occurs in other portions of the nephron.

Acetazolamide↗
PubMed1983

Seasonal and daily plasma corticosterone rhythms in American toads, Bufo americanus.

Concentrations of corticosterone were measured in the plasma of American toads, Bufo americanus, on a seasonal basis using a radioimmunoassay technique. Two populations of toads, maintained under different light conditions, were monitored to observe the effects of photoperiod on the seasonal rhythm of plasma corticosterone. Under a natural photoperiod toads demonstrated a rhythm consisting of a spring peak and a fall peak in corticosterone concentration. Toads maintained under a 12L:12D photoperiod all year round demonstrated a similar rhythm with peaks in the spring and fall. This suggests that an endogenous (circannual) rhythm of corticosterone may be playing an important role in the seasonal change of overt behavior and physiology of Bufo americanus. A daily rhythm of corticosterone was also detected in toads when blood samples were taken every 4 hr. When compared to a previously published circadian rhythm study of locomotor activity, the surge in corticosterone concentration for the day occurred at 1730 just prior to the peak in locomotor activity.

Animals↗
PubMed1990

Changes in plasma levels of gonadotropins and sex steroids in the toad, Bufo japonicus, in association with behavior during the breeding season.

Plasma levels of gonadotropins and sex steroids in Bufo japonicus were surveyed at different behavioral stages during the breeding period in March. All hormone levels were lower in torpid toads, which were found underground 1 week before the start of the breeding migration, than in active toads in the breeding season, although the levels were higher than those in the other months. LH and FSH levels in toads migrating toward a breeding pond were still low or only slightly elevated. Gonadotropin levels increased markedly when toads arrived at the pond and were highest for individuals actually in the pond. These LH and FSH surges occurred in both sexes, although peak levels were higher in females. Gonadotropin levels decreased in individuals of both sexes when they left the pond after breeding ("landed" individuals). LH and FSH levels in solitary males were significantly lower than those in amplexing males. In females, there was no significant difference in the level of any hormone between amplexing and solitary individuals. Circulating testosterone increased to the highest level in migrating males; this elevated level was maintained after arrival in the pond. DHT in males showed a pattern similar to that of testosterone. Both androgens decreased to the lowest level in landed males. In females, estradiol was dominant during migration, while progesterone became dominant shortly after arriving near or in the pond. This progesterone surge coincided with the LH surge. Androgen may stimulate amplexus behavior in male toads, and then amplexus seems to induce the LH surge that is necessary for spermiation. In females, however, amplexus seems to have no direct correlation with the gonadotropin surge, and the LH surge may induce the progesterone surge which causes ovulation.

Animals↗
PubMed1992

Digitalis-like compounds in the toad Bufo viridis: tissue and plasma levels and significance in osmotic stress.

Digitalis-like compounds (DLC), constituents of animal tissues, are possible regulators of the Na+, K(+)-ATPase implicated in water and salt homeostasis. The distribution of DLC in the toad (Bufo viridis) was determined following methanol extraction and partial purification. DLC highest levels were found in the skin but it was also detected in the plasma and many internal organs. Short term (hours) exposure of the toad to hypertonic shock (1.5% NaCl) induced an increase in plasma osmolarity due to an increase in Na+ and Cl- levels. This treatment induced a transient, three fold, increase of DLC levels in the brain and transient reduction of its levels in the ventral skin. Acclimation of the toads to burrowing conditions for six weeks resulted in an increase in plasma osmolarity due to a large increase in plasma urea with a small increase in ion concentrations. Under these conditions DLC levels in the dorsal skin increased by 100% without alteration of its levels in the plasma, brain and ventral skin. DLC levels in the toad brain of control animals, showed a significant dependence on season, being highest in the summer and lowest in the winter. DLC levels in the skin peaked in May while the levels in the plasma were season independent. The changes in DLC levels induced by the short- as well as long-term perturbations in the animal environmental salinity together with the seasonal differences suggest that DLC in the toad is involved in water and salt homeostasis of these animals, but may also participate in other unknown functions.

Animals↗
PubMed1996

A key by which the toad's visual system gets access to the domain of prey.

Searching for principles that allow toads to distinguish between prey and nonprey, we wondered how the toad's prey-catching activity measured as R differs in response to changes in significant configurational stimulus features. Elongated shapes moving worm-like in the direction of their longer axes are preferred prey dummies; but a toad is not a worm detector, and a worm is not the unique prey-catching releaser. Considering the frequency distributions of R values, we show that the release of prey catching is in a specific manner sensitive to the relation between the extensions of an object parallel (xl1) and perpendicular (xl2) to its direction of movement. It is the xl1 and xl2 features-relating algorithm that provides the key (instruction) by which the toad's visual system gets access to the domain of potential prey in terms of configurational cues. This, within behaviorally relevant limits, largely invariant algorithm also holds for segmented stimuli. Further investigations show that this principle of object discrimination is not due to experimental procedures but emerges as a species-common property, of which different toad species take advantage in a species-specific manner. Neurobiological correlates are discussed.

Animals↗
PubMed1982

Stimulation of adenosine 3',5'-monophosphate formation in mononuclear leukocytes by toad venoms.

The content of adenosine 3',5'-monophosphate in human mononuclear leukocytes was enhanced 3-5-times by venoms obtained from African toad (Bufo africanus), American toad (Bufo americanus), Colorado river toad (Bufo arenarum) and Marine toad (Bufo marinus) at 25 micrograms/ml for 5 min of incubation at 37 degrees C. The maximum stimulation was observed after 1-5 min of incubation. The half-maximal stimulation was observed at 0.1 micrograms/ml venom obtained from Colorado river toad (Bufo arenarum). The increased content of adenosine 3',5'-monophosphate in the mononuclear leukocytes persisted without significant change for at least 30 min of incubation at 37 degrees C.

1-Methyl-3-isobutylxanthine↗
PubMedDate not supplied

Characterization of cholecystokinin receptors in toad retina.

The binding characteristics, structure, and pharmacologic properties of a cholecystokinin binding protein in toad retinal membranes have been studied. In competition binding studies using 125I-CCK-8, toad retinal membranes exhibited a high affinity binding site having a Ki50 of 1.5 nM using CCK-8 as competitive ligand. The relative potencies of CCK-related peptides in inhibiting radioligand binding were caerulein greater than gastrin II approximately equal to CCK-8 approximately equal to CCK-33 greater than CCK-8-DS approximately equal to gastrin I. L-364,718, a potent inhibitor of peripheral CCK receptors, was ineffective at competition binding at concentrations up to 1 microM; dibutyryl cyclic GMP was modestly effective at competing (KD approximately 10 mM). Covalent binding of 125I-CCK-33 to toad retinal membranes using chemical cross-linkers or UV irradiation resulted in the labeling of a major Mr 62,000 protein and the intermittent labeling of minor components of Mr 105,000 and Mr 40,000 as determined by SDS-PAGE and autoradiography. The binding of 125I-CCK-33 to retinal membranes and the concomitant labeling of the Mr 62,000 component was specifically inhibited by CCK-8 (KD approximately 1.5 nM). Reduction of membranes with DTT abolished specific binding of 125I-CCK. SDS-PAGE analysis of affinity cross-linked membranes under non-reducing conditions revealed that the Mr 62,000 protein migrated with an apparently lower molecular weight. These results suggest that the Mr 62,000 CCK binding protein in the toad retina contains an intramolecular disulfide bond(s). The Mr 62,000 protein was retained on a wheat germ agglutinin-agarose column and eluted with N-acetyl D-glucosamine, suggesting the glycoprotein nature of this protein. Digestion of the Mr 62,000 protein with neuraminidase together with O-glycanase resulted in a discrete product of Mr approximately 60,000. These results indicate that the Mr 62,000 protein is a glycoprotein with O-linked oligosaccharide chains. Taken together, these data indicate that the CCK receptor in toad retina has a distinct structure compared to that described in rat pancreas or brain. It will be important to establish whether this difference is reflected in differences in signal transduction mechanisms.

Animals↗
PubMed1985

Osmoregulation of the cane toad, Bufo marinus, in salt water.

Adult cane toads, B. marinus, survived in salinities up to 40% sea-water (SW). Pre-exposure to 30, then 40% SW, increased the survival time of toads in 50% SW. Plasma from toads acclimated to salt water is hyperosmotic to the environment--a result of increased plasma sodium, chloride and urea concentrations. When toads were placed in tap-water and 20% SW, all significant changes to plasma sodium, chloride, urea and osmotic pressure occurred within the first 2 days of exposure. When toads were placed in 30 and 40% SW environments, the increases in plasma sodium and chloride concentrations occurred within the first 2 days of exposure while urea and total osmotic pressure continued to rise until some time between 2 and 7 days exposure.

Animals↗
PubMed1986

Plasma osmolality, urine composition and tissue water content of the toad Bufo viridis Laur. in nature and under controlled laboratory conditions.

The compositions of plasma and urine were studied in toads (Bufo viridis) which were collected from three locations in Israel, and compared with toads which were kept under constant laboratory conditions for nearly 2 years. Plasma osmolality was rather constant (over 310 mOsm kg-1 H2O) during the whole year in the active toads. Urea was the most variable osmolyte in the plasma, and accounted for the higher osmolality in southern population. Urine osmolality fluctuated in a circannual fashion both in freshly captured and in the toads under constant laboratory conditions. Water content of the tissues was constant throughout the year, independent of the plasma osmolality. It is concluded that high plasma urea concentration and the excretory system (kidneys and the urinary bladder) are important in sustaining constant plasma osmolality in active toads. Both mechanisms change annually and form the basis for the high terrestriality of this species.

Animals↗
PubMed1992

Effects of angiotensin II and bladder condition on hydration behavior and water uptake in the toad, Bufo woodhousei.

1. Water absorption response (WR) behavior and water weight gain were examined in hydrated toads, Bufo woodhousei, treated with angiotensin II (AII) or with a control Ringer's solution. The effects of urinary bladder condition (ad lib. bladder urine or empty bladder) were examined concurrently. 2. Toads treated with AII (100 micrograms/100 g body weight), spent more time in WR posture and absorbed more water than Ringer's-injected toads. 3. Toads with empty bladders maintained WR posture for longer periods of time and gained more weight than toads whose bladders were not emptied. 4. The effects of AII and bladder urine on water absorption by B. woodhousei appear to be separate and additive.

Angiotensin II↗
PubMed2004

Temperature may influence and regulate NF-YB expression in toad oocyte.

Fully grown oocytes derived from Bufo gargarizans maintained at relatively low temperatures (4 degrees C, LTE-oocytes) acquire the competence to resume normal meiosis. In contrast, fully grown oocytes derived from toads maintained at relatively high temperatures (28 degrees C, HTE-oocytes) never acquire maturation competence. By suppression subtractive hybridization, we obtained 18 ESTs preliminarily thought to be preferentially expressed in LTE-oocytes; of these, TS1-4 shared homology with the human and mouse NF-YB genes. We cloned the full-length toad NF-YB gene by RACE and identified three alternatively spliced transcripts: tNF-YB1, tNF-YB2, and tNF-YB3 (GenBank Accession Nos. AY442015, AY442016, and AY442017, respectively). Toad NF-YB was differentially transcribed and translated in LTE-oocytes versus HTE-oocytes, likely resulting in differential CCAAT-binding and/or transcriptional activity of NF-Y. Furthermore, toad cyclin B2 was differentially transcribed at high and low temperatures. Taken together, this report of the differential expression of toad NF-YB at different temperatures is the first evidence that temperature may influence and regulate NF-YB expression.

Alternative Splicing↗
PubMed2006

Metabolic measures of male southern toads (Bufo terrestris) exposed to coal combustion waste.

Southern toads (Bufo terrestris) are found in coal fly ash collection basins associated with coal-burning electrical power plants. These basins contain large amounts of trace metals and organisms found in these basins are known to accumulate large quantities of metals. Studies on a variety of organisms exposed to trace metals found that they experience a significant increase in standard metabolic rate. We experimentally exposed southern toads to metal-contaminated sediment and food and measured changes in standard and exercise metabolic rates as well as changes in body, liver and muscle mass, blood glucose, and corticosterone. We found that toads exposed to trace metal contamination gained significantly less mass (18.3 %) than control toads (31.3%) when food was limited and experienced significantly decreased RQ after exercise. However, contaminated toads did not experience changes in standard (x control = 0.114 +/- 0.016 mL O2 g(-1) h(-1); x (ash)=0.109 +/- 0.013 mL O2 g(-1) h(-1), p = 0.08) or exercise metabolic rates (x control = 0.53 +/- 0.06 mL O2 g(-1) h(-1); x ash =0.44 +/- 0.04 mL O2 g(-1) h(-1), p = 0.47) plasma glucose levels (p = 0.6), and hepatic or muscle percentage indices (p > 0.2 in all cases) whether food was limited or not.

Animals↗
PubMed2005

Studies on the poisonous skin secretion of individual red bellied toads, Melanophryniscus montevidensis (Anura, Bufonidae), from Uruguay.

Toads belonging to the genus Melanophryniscus contain toxic alkaloids in their skin. From six locations in south-eastern Uruguay 81 specimens of Melanophryniscusmontevidensis were collected. In whole animal methanolic extracts of individual specimens, alkaloids of the pumiliotoxin (PTX) group and indolizidines were identified by gas chromatography/mass spectrometry; the predominant component PTX 251D was assayed quantitatively. The PTX-content of the various toad populations was found to be highly variable among individual specimens as well as among the populations. Very high levels of PTX 251D were detected in toads of the western part of the collection area, whereas very low levels of this alkaloid were assayed in toads near the Brazilian border. Remarkably high concentrations of the non-alkaloid hydroquinone were found to be present in all toads. The analysis of extracts from 125 arthropod samples (Arachnida and Insecta, including termites, ants and beetles), which may represent a potential food source, revealed no alkaloids of the PTX group.

Alkaloids↗
PubMed2003

Cloning of cDNAs encoding the three pituitary glycoprotein hormone beta subunit precursor molecules in the Japanese toad, Bufo japonicus.

Complementary DNAs encoding precursor molecules of the beta subunits of three pituitary glycoprotein hormones (LH, FSH, and TSH) of the Japanese toad (Bufo japonicus) were isolated and sequenced. Unexpectedly large numbers of single nucleotide substitutions were found in all three beta subunit cDNAs. The eight isolated LH beta precursor cDNA clones were classified into six forms of nucleotide sequence, with four nucleotide substitutions each in the apoprotein coding region and in the 3' untranslated region (UTR). In the deduced amino acid sequence, the LH beta subunit showed two forms with a single amino acid substitution. The seven isolated FSH beta subunit cDNAs were classified into two forms, which differed from each other at 11 positions in the 3' UTR. The six isolated TSH beta subunit clones were classified into four forms with 2 and 5 nucleotide substitutions in the signal peptide and apoprotein coding regions, respectively. However, all the substitutions in the apoprotein coding region were silent. The substitution in the signal peptide coding region could produce three forms of signal peptide. Amino acid sequence comparison revealed that the toad LH beta subunit is more similar to the fish GTH II beta subunit than to mammalian and avian LH beta subunits. We found that the toad LH beta subunit molecule is a partial chimera of LH and FSH; amino acid residues located in 36th to 42nd and 96th to 99th are identical or similar to those of not LH- but FSH-beta subunit in mammalian, whereas it is more similar to LH- than FSH-beta subunit in total. We also found that the toad FSH beta subunit is more similar to the fish GTH II beta subunit than to the fish GTH I beta subunit and that the toad TSH beta subunit is more similar to tetrapod TSH beta subunits than to fish TSH beta subunits.

Amino Acid Sequence↗