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PubMed1991

Stimulation of membrane phospholipid metabolism by agents that increase acidification in toad urinary bladder.

Previous reports have indicated that metabolic acidosis stimulates H+ excretion, and this excretion is accompanied by an increased turnover of phospholipids (PL) in toad urinary bladder. The purpose of this experiment was to determine if other known stimulators of H+ excretion [insulin, deoxycorticosterone acetate (DOCA), epinephrine, parathyroid hormone, and CO2] might also stimulate PL turnover in the toad urinary bladder. Quarter bladders from normal toads were removed, weighed, and then incubated with [32P]orthophosphate for 2 hr at 25 degrees C. PL were extracted, separated, and detected using thin layer chromatography and autoradiography, and quantitated by liquid scintillation counting. Results were expressed in cpm (100 mg bladder)-1 (hr)-1. One quarter bladder received insulin (100 milliunits/ml), DOCA (10(-6) M), epinephrine (50 mM), parathyroid hormone (100 micrograms/ml), or 5% CO2 during the incubation, whereas the paired quarter bladder received no treatment. Phosphatidylcholine (PC) and phosphatidylinositol turnover were increased by insulin (P less than 0.025 and less than 0.05, respectively). DOCA had no effect on PL turnover, but stimulated the percentage fraction of PC (P less than 0.05) expressed as percentage fraction of total lipids. Five percent CO2 in the bath resulted in an increased rate of turnover of the PL fractions phosphatidylinositol (P less than 0.05), and the phosphatidic acid plus phosphatidyl-serine (P less than 0.01). Epinephrine and parathyroid hormone were both without effect on PL metabolism. We conclude that insulin, DOCA, and CO2 may stimulated H+ excretion in toad bladder in part by increasing turnover of membrane PL, PC, and phosphatidylinositol, and in the case of CO2, phosphatidic acid plus phosphatidylserine as well, but not PC.

Animals↗

PubMed1999

Mucormycotic dermatitis in captive adult Wyoming toads.

During late May 1995, 50 adult captive endangered Wyoming toads (Bufo baxteri) were brought out of hibernation. Approximately 3 to 10 days after hibernation emergence, all toads were hormonally induced to breed, and paired. Each pair was placed in their own breeding tank. Four toads developed clinical signs of disease which included lethargy and multiple (4 to 12) small (2 mm) raised hyperemic nodules with white fuzzy caps on the ventral skin. The condition progressively worsened until death occurred, within 3 to 6 days. Mycotic dermatitis caused by Mucor sp. was diagnosed in the four toads through histology and isolation of the organism. This is the first case report of a Mucor sp. causing a fatal dermatitis in an amphibian without significant inflammatory response and without systemic involvement.

Animals↗

PubMed2002

Effect of thyroxine on some digestive enzymes of the adult male toad, Bufo melanostictus.

Thyroxine is known to play an important role during the developmental process of amphibians. The present work is designed on the hypothesis that a functional relationship exists between the thyroid gland and digestive physiology in the adult toad. Three doses of thyroxine (25 microg/100 g; 50 microg/100 g; 100 microg/100 microg body weight) were orally administered daily to the adult male toad, Bufo melanostictus and changes in -amylase, pepsin, trypsin, and esterase activities of the different zones of the digestive tract, pancreas and liver were observed. The observations were made on 7, 15, and 30 days of thyroxine treatment. The result shows that the dose of 25 microg increases the enzyme activities in the gastrointestinal tract, liver and pancreas of 7 days treatment. However, prolonged treatment with all the doses shows little effect on these enzymes. The pattern of changes in the enzymatic activities in the digestive tract of the adult male toads show more advanced compartmentalization than that of fishes. It is also found that site-specific enzyme production is not pronounced in this amphibious vertebrate. It is proposed that a positive functional relationship between thyroxine and digestive enzymes exists in the adult male toad.

Animals↗

PubMed1992

[A voltage-dependent potassium channel of outward rectifier type in plasma membrane of oocyte from toad, Bufo bufo gargarizans].

Membrane properties of the fully-grown oocytes from toad, Bufo bufo gargarizans, were studied by using voltage-clamp technique. It was found that a sustained outward current was elicited by membrane depolarization to -30 mV or more positive value. The increase of the current was nearly proportional to the degree of depolarization. The peak value of the current ranged 2-5 microA at a membrane potential of 20 mV in oocytes from different toads. The current was inhibited by antagonists of potassium channel, TEA and 4-AP. The concentration of TEA capable of inhibiting half of the current was 2.6 mmol/L. Chloride channel antagonist 9-AC (2.5 mmol/L) had no effect on the current. Triple the extracellular calcium concentration did not show any effect either. The reversal potential of the current varied with an increase of 47.3 mV per decade change of the extracellular potassium concentration. Changing extracellular concentration of sodium or chloride did not shift the reversal potential. It was concluded that the outward current was a voltage-activated potassium current. The voltage-dependent potassium current decreased after treatment of the oocytes with progesterone to a state of maturation. A large decrease of the current (to about 1/20 of the control) occurred to the oocytes obtained from hibernating toads while a less striking decrease of the current (to about 1/3 of the control) was observed in the oocytes from toads all year round reared at 25-30 degrees C.

Animals↗

PubMedDate not supplied

Influence of nitric oxide on histamine and carbachol-induced gastric Acid secretion in the common african toad - bufo regularis.

The study aimed to determine the influence of nitric oxide (NO) on the action of histamine and carbachol on acid secretion in the common African toad - Bufo regularis. Gastric acidity was determined by titration method. The acid secretion was determined when nitric oxide was absent following administration of NO synthase inhibitor; N-nitro-L-arginine methyl ester (L-NAME) and when nitric oxide was in excess by administration of exogenous NO donor, sodium nitroprusside (SNP). Histamine or carbachol increased acid secretion in the toad. Acid output increased from 0.32 +/- 0.04 mEq/15min to 0.56 +/- 0.08 and 0.61 +/- 0.05 mEq/15min for histamine and carbachol respectively [P < 0.05]. Pretreatment of the toad with L-NAME produced further increases in histamine (0.62 +/- 0.06 mEq/15min) or carbachol (0.74 +/- 0.06 mEq/15min) induced acid secretion respectively. SNP however, completely abolished the acid secretion stimulated by either histamine or carbachol. It was therefore concluded that nitric oxide has a negative influence on the histamine or carbachol-stimulated acid secretion in the toad - Bufo regularis.

Journal Article↗

PubMed1990

The action of progesterone on sodium transport of isolated toad skin. Further evidences of a metabolic action.

To understand the mechanism of progesterone inhibition of the sodium influx in the toad skin, the effects of the hormone on the active sodium transport and oxidative metabolism of the transporting cells were examined. A direct relationship was observed between the initial value of the short circuit current and the sensitivity of a given skin toad preparation to progesterone. Progesterone had a higher effect on the sodium potential than on any other parameter of an equivalent electrical circuit of toad skin. Direct measurement of oxygen consumption indicated that progesterone can act as a blocking agent of the respiratory chain of the sodium transporting cells of toad skin.

Animals↗

PubMed1979

Studies on the erythropoietic effect of plasma from anemic toads both with and without testis.

Injection of plasma from experimentally induced anemic toad with intact testis increases erythropoiesis in starved toads evidenced by the increase of red blood cell, hemoglobin and hematocrit, whereas the plasma of castrated and phenylhydrazine-HCl treated anemic toad failed to do so. It can be suggested that the erythropoiesis stimulating factor (ESF) is produced from the testis of toad and the production of this factor (ESF) was found to increase during anemia in an attempt to correct the anemic condition.

Anemia↗

PubMed1981

Hypophysis effect on the topographical distribution of toad oviduct mucoprotein components.

In the present work the effects of the hypophysis hormones on oviduct mucoprotein components distribution patterns were studied. Remarkable changes after treating the toad with hypophysis injections were apparent. The distribution pattern for hexose, sialic acid, hexosamine and phosphate from 18 hours hypophysis treated toads were found to be identical with those obtained from preovulatory period animals. On the other hand, the levels for mucoprotein components from hypophysis treated animals were found to be approximately one-half or more higher than those obtained from postovulatory period toads. Otherwise, hypophysis treatment of the toads in preovulatory period had not effect on the levels and distribution patterns of mucoprotein components. These results suggest that hypophysis hormones are involved in the increase of the oviduct secretory activity.

Animals↗

PubMed1994

Opioid binding in giant toad and goldfish brain.

Opiate receptor expression in phylogenetically different species has played an important role in the study of opioid receptor pharmacology. Total opioid binding measured with the nonselective ligand 3H-diprenorphine reveals a Bmax of 21.7 +/- 1.37 fmol/mg tissue wet wt and a KD of 0.17 +/- 0.03 nM in Bufo marinus (giant toad), as well as a Bmax of 18.17 + 0.41 fmol/mg tissue wet wt and a KD of 0.47 +/- 0.18 nM in Carassius auratus (goldfish). Despite the similar levels of 3H-diprenorphine binding, the composition of binding subtypes in the two species differs. Approximately 30% of total binding corresponds to mu receptors in both species, whereas neither kappa 1 nor delta binding can be detected. However, the remaining 70% of binding differs between the toad and goldfish. In the toad, the non-mu binding corresponds to kappa 2 sites, whereas in the goldfish, the non-mu binding corresponds to kappa 3 sites. The sites can be distinguished biochemically, as well as pharmacologically. After affinity labeling the sites with 3H-NalBzoH, the retention times on an ion-exchange column differ for the peaks of kappa binding in the two species. Although Bufo marinus (giant toad) and Carassius auratus (goldfish) brains express kappa and mu opioid binding, the kappa subtypes in these two species differ.

Animals↗

PubMed1996

Initiation and post-initiation chemopreventive effects of beta-carotene in toad liver carcinogenesis.

Hepatocellular carcinoma were recognized in the toad, Bufo viridis, in 14 cases out of 50 by injection of 1 mg 7,12 dimethylbenz(a)anthracene (DMBA)/toad twice/week for 12 weeks. In contrast, toads treated with DMB at the same dose level and beta-carotene (betaC), 0.05 mg (3 hr prior to the carcinogen)/toad, twice/week for 12 weeks showed no tumor incidence. However, betaC at the same dose level was less effective when administered 3 hr. after the carcinogen (DMBA). In 8 cases out of 50 cases neither tumor growth nor neoplastic changes were observed in toads treated with betaC alone or olive oil. It is concluded that betaC completely blocked hepatocarcinogenesis in toads when given 3 hr. before initiation.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed1977

Toads as natural reservoir of salmonella.

The investigation conducted on 329 toads for the presence of Salmonella, Arizona and Edwardsiella revealed that 36.7% toads were intestinal carriers of Salmonella. None of the toads proved positive for Arizona and Edwardsiella. The Salmonella population (S. brijbhumi) in the intestines of 5 of the 27 toads ranged from 1.5 X 10(9) to 2.5 X 10(10) organisms per gram of faecal matter. Some of the serotypes viz: S. tel-hashomer, S. brijbhumi and S. goverdhan were recovered more frequently than others. An unnamed serotype (4, 12, 27:k:-) and S. goverdhan var 1 + (1,9,12:k:1,6), hitherto undescribed in the literature, have also been reported.

Animals↗

PubMed1965

EFFECT OF ALKALI METAL CATIONS ON THE POTENTIAL ACROSS TOAD AND BULLFROG URINARY BLADDER.

Isolated urinary bladders of the bullfrog (R. catesbeiana) and the toad (B. marinus) were mounted in an Ussing chamber. Potential differences up to 114 mv were observed in bullfrog bladder when the mucosal surface was bathed in dilute Na(2)SO(4) and the serosal surface in sulfate Ringer's. In experiments with bullfrogs, K was used to replace Na in the mucosal solution and Na was used for K in the serosal solutions. The selectivity was judged in terms of the relative effectiveness of the replacement cation in maintaining the bladder potential. In experiments with toads, K and Rb were equally poor replacements for Na at the mucosal border, while Rb was a good replacement for K at the serosal border. Li in the mucosal solution appeared to depress the potential in part irreversibly. At the serosal border, Li was a partially effective substitute for K, more so than was Na. However, both were poor replacements compared to Rb. The mucosal surface of the urinary bladder of both frog and toad appears to be Na-selective and the serosal surface appears to be K-selective, consistent with the Koefoed-Johnsen-Ussing model for frog skin.

Animals↗

PubMed1964

INNERVATION OF THE LARGE INTESTINE OF THE TOAD (BUFO MARINUS).

The morphology, physiology and pharmacology of the innervation of the toad (Bufo marinus) large intestine have been studied. The large intestine can be divided into the regions colon, rectum and cloaca, on morphological grounds, but acts as a unit in response to nerve stimulation. Of the right and left nerves, each appears to supply the entire large intestine. Autonomic innervation of the large intestine of Bufo marinus is as follows: (1) The 9th and 10th spinal nerves (pelvic) contain predominantly excitatory preganglionic cholinergic fibres, but some inhibitory adrenergic fibres are also present in most preparations. (2) The splanchnic nerves contain inhibitory postganglionic adrenergic fibres from the 3rd to 5th sympathetic ganglia, and a small number of excitatory cholinergic fibres. The pathway of adrenergic inhibitory fibres to the large intestine alongside the posterior mesenteric artery as seen in mammals is rarely present in the toad. Several nonspecific actions of autonomic drugs on the large intestine are discussed. The functional organization of the autonomic innervation of the toad large intestine is similar to that in mammals, that is the large intestine is controlled by antagonistic cholinergic and adrenergic nerves. However, the separation of these two types of nerve fibres into anatomically distinct nerves does not appear to be as complete as in mammals. It is suggested that inhibitory autonomic control of the alimentary canal in vertebrates first appears in the hind-gut region.

Acetylcholine↗

PubMed1999

Elevated plasma osmotic concentration stimulates water absorption response in a toad.

The water-seeking behavior (WR) of toads (Bufo viridis) was investigated. Fully hydrated toads that are allowed free choice of wet or dry filter paper voluntarily and spontaneously select to sit on water-soaked paper at a regular frequency during trials. Dehydration of bladder-emptied toads by 14% elicits WR in all animals. Injection of aldosterone or angiotensin-I reduced the dehydration threshold to 7% weight loss. WR frequency increased when plasma osmolality was elevated by injection of NaCl or other solutes (both ionic and non-ionic). Only urea, to which cell membranes are highly permeable, was the exception that did not produce this response. The increase in WR frequency induced by elevated plasma osmolality was augmented by injection of aldosterone or angiotensin-I. In vivo water uptake, measured in a water bath, was increased by an NaCl or oxytocin injection, but not by aldosterone. It is concluded that elevated plasma osmolality induces an increase in WR frequency that is separate and prior to the water uptake process. Different hormones are involved in each step.

Aldosterone↗

PubMed1997

Morphological features of the retinal pigment epithelium in the toad, Bufo marinus.

BACKGROUND: The toad's (Bufo marinus) retinal pigment epithelium (RPE) has been used in many studies as a model for understanding its role and interaction with the neural retina (NR). In this study, the fine structure of the RPE is described. METHODS: In vitro RPE-chorion preparations from the toad were studied after separation from the NR. Tissues were processed for light and electron microscopy. RESULTS: The RPE consists of a single layer of cells joined by a series of tight junctions forming part of the blood-retina barrier. These cells had minimal infolding of the basal membrane and numerous microvillous processes extending from the apical surface and surrounding photoreceptor outer segments. Internally, RPE cells display a large nucleus, numerous mitochondria located at the basal membrane, extensive smooth endoplasmic reticulum, myeloid bodies, phagosomes, scattered polysomes, large lipid droplets and melanin granules. Rough endoplasmic reticulum is relatively scarce within these cells. The choriocapillaris consists of large blood vessels facing Bruch's membrane. Bruch's membrane is typical of other vertebrates, being pentalaminate over the whole retina. The endothelium of the choriocapillaris facing Bruch's membrane is typically very thin but shows few fenestrations. CONCLUSIONS: Due to the ultrastructural similarities of toad RPE with that of mammals, it can serve as an excellent model for the study of retinal detachment/reattachment and their cellular and molecular mechanisms.

Adaptation, Ocular↗

PubMed1982

Degranulation of endothelial specific granules of the toad aorta after treatment with compound 48/80.

Incubation of isolated toad aortas in Ringer solution containing compound 48/80, a histamine liberator, resulted in marked degranulation of endothelial specific granules. Since incubation of these vessels in Ringer solution only did not show significant morphologic changes in these granules, these findings suggest that the degranulation was induced by histamine release from the granules, as in the case for mast cell degranulation, and that endothelial specific granules are a storage site of histamine in the toad aorta. The present morphologic data were supported by preliminary chromatography, which showed appreciable concentrations of histamine in the granule-containing pellets of subcellular fractions of homogenized toad aortas.

Animals↗

PubMed1983

The blood-testis barrier in the toad (Bufo arenarum Hensel): a freeze-fracture and lanthanum tracer study.

Intercellular junctions between Sertoli cells in the toad testis were studied by freeze-fracture and electron-opaque intercellular markers. These junctional specializations are characterized in thin sections by a series of focal fusions on the outer leaflets of both adjacent cell plasmalemmas, associated with bundles of fine filaments in the subjacent Sertoli cell cytoplasms. However, the wide subsurface cisterna of the endoplasmic reticulum, a component constantly associated with Sertoli cell junctions in mammals, is absent in the toad. The intravascularly injected lanthanum hydroxide, used as a tracer compound, gains access to the seminiferous tubules and surrounds spermatogonia and leptotene spermatocytes, but is persistently excluded from germ cells in later stages of development. This indicates that, as is the case in the mammalian testis, a permeability barrier to lanthanum is established which isolates all germ cells beyond leptotene spermatocytes. Freeze-fracture reveals the characteristic occluding junctions between Sertoli cells, but a variation in their geometric patterns was clearly observed in different regions of the toad seminiferous epithelium. The membrane-fractured faces of Sertoli cells embracing differentiating spermatids exhibit a deep junctional complex: up to 50 rows of particles between adjacent Sertoli cells separate these late germ cells from the periphery of the seminiferous tubules. Sertoli cells surrounding early germ cells generally exhibit, instead, a discontinuous, poorly developed network of interconnected rows of particles with few widely spaced strands. This seems to permit the percolation of the intercellular marker in areas of the seminiferous epithelium containing spermatogonia and leptotene spermatocytes.

Animals↗

PubMed1995

Histamine release from Weibel-Palade bodies of toad aortas induced by endothelin-1 and sarafotoxin-S6b.

BACKGROUND: Endothelin-1 (ET-1) and sarafotoxin-S6b (STX) induce a remarkable degranulation of Weibel-Palade (WP) bodies prior to the vasocontraction of toad aortas. As WP bodies play the role of a reservoir site of the histamine in the endothelial cells, there is the possibility that ET-1 and STX evoke the release of histamine from WP bodies of this vessel. METHODS: Histamine concentrations were assayed by high-performance liquid chromatography (HPLC) from the perfusate after being perfused with a solution containing ET-1 and STX. Each vessel was fixed and embedded for conventional electron microscopy and immunoelectron microscopy using antihistamine sera. RESULTS: The appreciable concentrations of histamine were assayed by HPLC from the perfusate after the toad aortas were perfused with a solution containing ET-1 and STX. The immunoelectron microscopy revealed that histamine immunoreactive gold particles in the WP bodies remarkably decreased in number in the treated samples when compared to the control ones. Our immunoelectron micrographs indicated that the release of histamine from the endothelial cells occurred in association with the degranulation and the exocytosis of the WP bodies after treatment with ET-1 and STX. CONCLUSIONS: The present study clearly shows that ET-1 and STX induce the histamine release from WP bodies of the toad aortas by means of HPLC and immunoelectron microscopy. Histamine discharged from the WP bodies may be involved in the vasocontraction evoked by ET-1 and STX.

Animals↗