Morphology of the oral disc of Bufo bufo (Salientia: Bufonidae) tadpoles

The fully developed oral disc of the tadpole of Bufo bufo consists of dorsal and ventral labia bearing, respectively, two and three ridges bearing numerous horny denticles, a horny beak provided with jaw sheath serrations, and large lateral papillae that are borne by two cutaneous plicae. As development progresses toward metamorphosis, these structures gradually regress until they disappear. Each cusped clavate labial denticle adheres, by means of a thin peduncle, to a similar labial denticle fixed in the lip and formed by a group of three or four cells that keratinize gradually and thus present remarkable differences in their morphology. Once all the cells of a group have been converted into horny tissue, the denticle sheds and is replaced by the underlying one. The beak serrations also are horny structures; each consists of a columnar band of cells which undergoes a gradual keratinization. The horny cells that detach themselves at intervals, being replaced by those of the underlying anlagen. The labial denticles and the beak serrations keratinize in two distinct ways. In the former, the desmosomal filaments appear to play an important role whereas, in the latter, the keratin seems to be synthesized “ex novo” by the ribosomes.     

Toxin depletion has no effect on antipredator responses in common toad (Bufo bufo) tadpoles

Antipredator responses often involve changes in several phenotypic traits and these changes interactively influence fitness. However, gaining insight into how the overall fitness effect of the overall response comes about is notoriously difficult. One promising avenue is to manipulate a single defensive trait and observe how that modifies fitness as well as the expression of other inducible responses. In chemically‐defended animals, toxins are likely to be costly to produce but it is still unknown how their depletion influences other characteristics. In the present study, we artificially depleted bufadienolide toxin stores in common toad (Bufo bufo) tadpoles, and assessed the effect of this with respect to the interaction with predator presence and limited food availability. We found that toxin depletion in tadpoles did not significantly affect any of the measured life‐history traits. Tadpoles in the predator treatment exhibited an elevated development rate, although this was only apparent when food availability was limited. Also, body mass at metamorphosis was lower in tadpoles exposed to chemical cues indicating a predation threat and when food availability was limited. These results provide evidence that, in larval common toads, the expression of inducible defences may incur fitness costs, whereas chemical defences are either expressed constitutively or, if inducible, elevated toxin production has negligible costs.     

Influence of thyroid hormone on the development of preoptic-hypothalamic monoaminergic neurons in tadpoles of Bufo bufo japonicus

The effect of thyroid hormone on the development of diencephalic monoaminergic neurons was studied in tadpoles of Bufo bufo japonicus. Monoamine-containing neurons in the preoptic recess organ (PRO) appeared later than those in the paraventricular organ (PVO) and nucleus infundibularis dorsalis (NID). After deprivation of thyroidal primordium no fluorescent neurons developed in the PRO. Development of monoaminergic neurons in the PVO and NID was not affected by thyroidectomy. Thyroxine treatment brought about the fluorescent neurons in the PRO of the thyroidectomized tadpoles. Fluorescent terminals in the median eminence became conspicuous around the capillaries which penetrated to the median emience, when the tadpoles reached late prometamorphic stage. In the median eminence of thyroidectomized tadpoles, the monoaminergic axon terminals did not develop. Thyroxine induced both the fluorescent terminals and the capillary penetration in the median eminence of the thyroidectomized tadpoles. In the tadpoles hypophysectomized at tail-bud stage, thyroxine induced neither the fluorescent terminals nor the capillaries in the median eminence.     

Behavioral reactions of Bufo bufo L. tadpoles to chemical excretion of adult toads and changes during larval development

The reactions of toad (Bufo bufo L.) tadpoles to chemical excretes of adults were studied. In double-choice test aquarium tadpoles demonstrated avoidance reactions (swimming away from the place of stimulus delivering, reduction time of spent in stimulus part of aquarium and reduction of movement activity). Efficiency of stimulus reduced when its concentration decreased. The thresholds of tadpoles sensitivity estimated according to the reduction of time spent in stimulus part of aquarium decreased in onthogeny: they were near 500 x 10(-5) g hour/l at 36-39 developmental stages and 500 x 10(-4) g.hour/l at 40-41 stages.     

Growth response of testis tissue to partial castration in toads, Bufo bufo bufo

The growth response of the remaining intact testis or testis fragments to partial castration was studied as a function of the duration of the postoperative period, the amount of testis mass excised, as well as the functional state of the testes at the time of operation. Excision of about 90% of the testis mass caused a growth response that increased from slight after eight weeks to pronounced after 14 weeks. After 14 weeks the growth response was slight tounilateral excision of 75% of a testie and pronounced to bilateral excision of 75% of each testis. Subtotal castration caused formation of new seminiferous tubules within the remaining testis tissue when the operation was performed early in the annual testis cycle, whereas the growth response late in the cycle was primarily caused by increased spermatogenetic activity within existing tubules. Partial castration stimulated oocyte formation within fragmented testes, but not in a remaining intact testis. Oocyte formation within a testis fragment was independent of the presence of the Bidder's organ.     

Ultrastructural analysis and cytochemical localization of atrial natriuretic peptide-stimulated guanylate cyclase in internal gills of Bufo bufo tadpoles.

In this study on the internal gills of the larvae of Bufo bufo we examined the ultrastructural features and, using cytochemical methods, showed the localization of guanylate cyclase in the presence of atrial natriuretic peptide. The gill apparatus consists of a series of arches each with a dorsal part or gill rakers with filtering and glandular functions. In the epithelium, cells were found that contain granular secretions similar to those atrial natriuretic factor-immunoreactive granules of larval Bufo arenarum gill rakers. The ventral portion of the gill arches is made up of gill tufts with a respiratory function. The cytochemical localization of the guanylate cyclase in the presence of exogenous atrial natriuretic peptide demonstrates that the internal gills of the larvae are an important target organ for the peptide and therefore, it is proposed that, at this level, the atrial natriuretic peptide carries out an important osmoregulatory role.     

Gonadal toxicity of short term chronic endosulfan exposure to male rats

Endosulfan was studied for its effect on rat testicular toxicity in relation to the enzymes of androgen biosynthesis, viz. 3 beta-hydroxysteroid dehydrogenase (EC 1.1.1.145, 3 beta-HSD) and 17 beta-hydroxysteroid dehydrogenase (EC 1.1.1.64, 17 beta-HSD); cytosolic conjugation enzyme, glutathione-S-transferase (EC 2.5.1.18); and testicular as well as serum testosterone levels at the dose levels of 2.5, 5.0, 7.5 and 10 mg/kg body weight fed orally for 7 and 15 days. Organ and body weights of the treated animals did not change significantly, however, the testicular protein contents were found to be increased appreciably after 7 days treatments. The activity profile of cytosolic conjugation enzyme showed much remained low during 7 days treatment, however, the two steroidogenic enzymes showed much individual variations in response to endosulfan treatments. An overall varied response with respect to testosterone biosynthesis and its secretion to serum was observed suggesting nevertheless, a profound hormonal imbalance caused by this insecticide to male gonads on short term chronic exposures.     

Ultrastructure of the paraphysis in Bufo bufo larvae

A study of the ultrastructure and function of the paraphysis in Bufo bufo larvae was carried out. The structure is a tubular-ramified gland made up of numerous tubules with monolayered epithelial walls surrounded by connective tissue and sinusoids. The epithelial cells secrete glycoprotein to contribute to production of the cephalorachidian fluid. The role of the paraphysis in the transport of fluids and electrolytes from the blood to the cephalorachidian fluid in regulation of ionic and osmotic homeostasis is discussed. © 1994 Wiley-Liss, Inc.     

Morphofunctional mitochondrial response to methylglyoxal toxicity in Bufo bufo embryos

Methylglyoxal (2-oxopropanal) is a reactive alpha-oxoaldehyde that can be formed endogenously mainly as a by-product of glycolytic pathway. It is a cytotoxic compound with significant antiproliferative properties as it can bind, under physiological conditions, to nucleic acids and proteins, forming stable adducts. We have recently shown that exogenous methylglyoxal (150-600 microM) is highly toxic for amphibian embryos where it produces, when added to the culture water, inhibition of cell proliferation in the early developmental stages, followed by severe malformations and strongly reduced embryonic viability. In this work we investigate the morphofunctional effect of methylglyoxal on the common toad B. bufo embryo mitochondria in order to verify if its dysmorphogenetic action might be also ascribed to impairment of mitochondrial functions. The mitochondria were isolated from embryos at the developmental stages of morula, neural plate and operculum complete and developing in the presence of 600 microM methylglyoxal. The results show that exogenous methylglyoxal is highly toxic at mitochondrial level, where it produces proliferation, swelling and membrane derangement. As a consequence, mitochondria from treated embryos show decreased oxidative phosphorylation efficiency, as indicated by the significant reduction both of the respiratory control index values and of the embryonic ATP content. On the basis of these data, it is possible that the methylglyoxal-induced embryonic malformations as well as the strongly reduced viability might be also ascribed to energy depletion.     

Developmental aspects of hexose metabolism in Bufo bufo

Due to the close correlation between glucose mobilization and utilization within animal tissues, in this paper, the stages of appearance of phosphorylase, glucose-6-phosphatase and hexokinase as well as the levels of some intermediates of glucose metabolism have been investigated during Bufo bufo development. Phosphorylase first appears at stage 13 and is dominant in the neural part of the embryo, but, after this stage, increases relatively more in the nonneural one. Hexokinase appears at stage 17 and glucose-6-phosphatase soon after. Phosphorylase appearance at stage 13 is correlated with an increase of lactate content in the embryo; this may indicate a metabolization of hexoses. On this basis, the subsequent appearance of hexokinase and glucose-6-phosphatase activities also seems coherent with hexose mobilization and utilization within embryo. No direct causative factor for the changes observed was evident.     

Permeability properties of the Bufo bufo bladder as affected by isoprenaline and vasopressin

Isoprenaline, a beta adrenergic agonist, strongly increases both transepithelial fluxes across the urinary bladder of Bufo bufo; this effect is dose dependent, 10(-6)M being necessary for the maximal action. This effect is less selective than that of vasopressin: the ratio J urea/J thiourea is 3.8 under isoprenaline and 30.4 under vasopressin treatment. Both hormones differently affect the permeability of a mainly liposoluble molecule, i.e. antipyrine: vasopressin increases antipyrine permeability, while isoprenaline decreases it. Moreover diethylpyrocarbonate treatment of the luminal membrane strongly inhibits vasopressin effect on urea permeability leaving unmodified that of isoprenaline. However, the actions of both hormones are not additive. These results allows to assume that the tissue has a feedback mechanism which inhibits other hormonal action while the bladder is stimulated by a particular hormone.     

The reaction of tadpoles of the common toad (Bufo bufo L.) to chemical signals from individuals of their own and other species

In behavioral experiments, toad tadpoles-recipients chose between two parts of test aquarium with chemical signals of donor tadpoles of the same or different (Rana esculenta) species. The youngest studied tadpoles (43-45 stages) preferred the part of the aquarium with chemical cues from sibs as compared to non-sibs. No reliable difference in "non-sibs: water" system was observed. In "tadpoles of the same species: tadpoles of the other species" system, recipient tadpoles chose "their" part of the aquarium. Hence, toad tadpoles revealed a trend to aggregate with their sibs and avoid R. esculenta toads in the presence of certain chemical cues and in the absence of visual signals.