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Temporal selectivity of evoked vocal responses of Batrachyla antartandica (Amphibia: Leptodactylidae)

The advertisement call of the leptodactylid frog Batrachyla antartandica from southern Chile consists of a train of brief percussive tone pulses whose energy is centred at about 2 kHz. To gain an understanding of the temporal features that are essential for call recognition, playback experiments were conducted with 11 males. Subjects were presented with a synthetic imitation of this signal and variants for which different temporal call parameters were modified systematically. The number of pulses, pulse rate and latency of evoked vocal responses (EVRs) to stimuli having high pulse repetition rates (i.e. 8 and 16 pulses/s) were significantly weaker relative to responses to stimuli having an equal number of pulses but lower pulse rates. A similar, non-significant tendency was observed for a series of stimuli with different pulse rates for which the total stimulus duration was held constant. EVRs also decreased significantly for stimuli having long pulse durations (i.e. 48 and 96 ms) relative to stimuli comprising shorter pulses. No significant differences were observed between EVRs to stimuli for which pulse rise and fall times were varied from 1-20 ms. Responses to calls comprising trains of 10 pulses were weaker compared with stimuli having fewer pulses per train. The selective EVRs of B. antartandica for different temporal parameters contributes to an understanding of the mechanisms involved in call recognition and stress the relevance of temporal processing of sound by males for the emergence of specific patterns of vocal behaviour in anurans.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour

Journal Article

Aromatase activity in larval gonads of Pleurodeles waltl (Urodele Amphibia) during normal sex differentiation and during sex reversal by thermal treatment effect.

Estrogens are involved in gonadal differentiation in birds and in reptiles with temperature-dependent sex determination, suggesting a key role for aromatase, the enzyme complex converting androgens to estrogens. Pleurodeles waltl is an amphibian with ZZ/ZW genotypic sex determination but gonadal differentiation is thermosensitive. The sexual phenotype is in conformity with the sexual genotype at ambient temperature (20 +/- 2 degrees), but ZW animals are sex-reversed in functional males when larvae are reared at 32 degrees from stage 42 to stage 54 (thermosensitive period). Histological sexual differentiation of gonads begins at stage 53. Aromatase activity was measured either in the gonadmesonephric complexes (stages 47 to 52) or in the gonads (stages 53 to 56) of ZZ and ZW larvae reared at ambient temperature or at 32 degrees and of ZW larvae shifted at stages 53, 55, or 56 from ambient temperature to 32 degrees for 48 hr. At ambient temperature, aromatase activity was detected, at low levels, in gonad-mesonephric complexes of both ZZ and ZW larvae at stages 47 and 50. At stage 52, it was significantly higher in ZW individuals than in ZZ ones. Then it remained low in gonads of ZZ males, whereas it markedly increased in gonads of ZW females. At 32 degrees, the gonad-mesonephric complexes (stage 52) and the gonads (stages 53 to 56) of ZW larvae had low aromatase activity similar to that in ZZ individuals. The exposure to 32 degrees for 48 hr of ZW individuals significantly decreased gonadal aromatase activity during the thermosensitive period (shift at stage 53) but not after the thermosensitive period (shifts at stages 55 and 56).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Purification and structural characterization of insulin from the lesser siren, Siren intermedia (Amphibia: Caudata).

Insulin has been isolated from an extract of the pancreas of a salamander, the lesser siren Siren intermedia, and its primary structure was established as: A-chain, Gly-Ile-Val- Glu-Gln-Cys-Cys-His-Asn-Thr10-Cys-Ser-Leu-Tyr-Gln-Leu-Glu-Asn-Tyr- Cys20-Asn, and B-chain, Val-Pro-Asn-Lys-Pro- Leu-Cys-Gly-Ala-His10-Leu-Val-Glu-Val-Met-Tyr-Phe-Val- Cys-Gly20-Asp-Arg-Gly-Phe-Phe-Tyr-Pro-Ser-Ser-Thr 30. Although those amino acid residues considered to constitute the receptor-binding region of insulin have been retained, siren insulin contains several substitutions (Gln-->Lys at B4, Ser-->Ala at B9, Ala-->Val at B14, Leu-->Met at B15, Leu-->Phe at B17, Pro-->Ser at B28, and Lys-->Ser at B29) of amino acid residues that are conserved in insulins from species of other amphibian orders. The biological activity of siren insulin was not investigated in this study but the substitutions at B28 (involved in dimer formation) and at B14 and B17 (involved in hexamer formation) may be expected to influence conformation and therefore biological potency. The data are consistent with the view that the Sirenoidea represent an early divergence from the ancestral stock of salamanders.

Amino Acid Sequence

A tandemly repeated DNA family originated from SINE-related elements in the European plethodontid salamanders (Amphibia, Urodela).

We have characterized a highly repetitive family, named Hy/Pol III, in the genome of the European salamanders Hydromantes (Plethodontidae). This family consists of short, tandemly repeated sequences organized in clusters, scattered through the genome as shown both by in situ hybridization to chromosomes and by Southern blot hybridization. The repeat unit is about 200 bp in length and it is a composite element since it contains a SINE-like retroposon with a tRNA structure, flanked by two short direct repeats. The whole element itself is bordered by two other direct repeats. The sequence data suggest that two elements, presumably derived from polymerase III transcripts, have been inserted one into the other, giving rise to the observed composite structure. During evolution the Hy/Pol III family was then amplified by tandem duplication at the DNA level. The inferred relationships between Hy/Pol III members from three representative species of the European Hydromantes suggests that a subfamily structure characterizes the evolutionary history of this family.

Animals

Optic nerves in plethodontid salamanders (amphibia, urodela): neuroglia, fiber spectrum and myelination.

In five species of lungless salamanders, family Plethodontidae, which all show highly developed visual abilities, the ultrastructure of the optic nerve was investigated and the total number of retinal ganglion cell axons, the percentage of myelinated axons, and the volume densities of glia and axons were determined. More than 80% of all axons were smaller than 0.4 micron and only 2-3% were larger than 0.8 micron. In individual nerves the degree of myelination varied between 1 and 9% which is in the range reported for other amphibian species. The miniaturized and highly paedomorphic species Batrachoseps attenuatus was an exception because only very few or even no myelinated axons were present in the nerve, which is unique among gnathostome vertebrates. The five investigated species had total numbers of axons ranging from 26,000 in Batrachoseps attenuatus to about 50,000 in Plethodon jordani. These numbers are the lowest found among vertebrates with an elaborated visual system. The amount of glial material in the optic nerve varied between 25 and 50%, with larger nerves possessing more glia than smaller ones. Ultrastructural analysis revealed that the optic nerve of each species contained both astrocytes and oligodendrocytes, although often in immature form. In Batrachoseps attenuatus the glia showed features of both astrocytes and oligodendrocytes which reflect an undifferentiated state.

Animals

The innervation of the uterus in Salamandra salamandra (L.) (Amphibia, Urodela). A morphological and biochemical study.

The innervation of the uterus in the ovoviviparous urodele Salamandra salamandra was studied. In whole mount preparations of the thin-walled uterus of pregnant females, a dense adrenergic network was demonstrated using a modified glyoxylic acid fluorescence technique. Based on vesicle type and cytochemical reactivity after chromate/dichromate fixation for electron microscopy at least two types of neural process were distinguished and classified as adrenergic and cholinergic. Both types are preferentially situated above or between the smooth muscles of the uterine tissue. Adjacent to the muscles in the walls of arterioles mainly adrenergic fibers are seen. Using high performance liquid chromatography with electrochemical detection in tissue homogenates of uterus a considerable amount of noradrenaline could be identified. The significance of the dense innervation is discussed with respect to the function of the uterus during pregnancy and birth.

Adrenergic Fibers

Gap junctions and impulse propagation in embryonic epithelium of Amphibia. A freeze-etching study.

Epithelium of amphibian embryos (Cynops orientalis, Xenopus laevis) was found in preceding experiments to generate and conduct impulses during a limited stage (26-37) of development . In order to elucidate the structural basis of impulse propagation, epithelial cells of four stages were examined by the freeze-etching method: (I) before and (II) during acquisition of conductivity; (III) when propagation was fully established, and (IV) when it was no longer present. Only few gap junctions (GJ) of small size were found in groups I and IV. GJ in epithelia of group III were increased in number and size, and appeared morphologically "coupled", i.e., with more loosely arranged connexons. the size of gap-junctional particles did not differ significantly between coupled and uncoupled stages. Zonulae occludentes seemed "leaky" in stage *, and "tight" in stages II-IV. Thus, the morphological characteristics of specialized junctions between "non excitable cells" correlated with the opening and closing of low resistance intercellular current pathways during embryonic development. Gap junctions in particular seem to form an essential link in the non-neural stimulus-response system, which may facilitate the mobility of the embryo during early phases of aquatic life before the reflex pathways have been established. Coupling and uncoupling of gap junctions may also play an important role in the regulation of cell differentiation and morphogenetic movement. The experimental model used in this study provides a useful tool for further investigations of structural correlates of gap junctional permeability under physiological conditions.

Animals

Some evidence for the ampullary organs in the European cave salamander Proteus anguinus (Urodela, Amphibia).

The multicellular epithelial organs in Proteus anguinus, which Bugnion (1873) assumed to be developing neuromasts, have been analyzed by light- and electron-microscopy. Their fundamental structure consists of single ampullae with sensory and accessory cells with apical parts that extend into the pit of the ampulla, and of a short jelly-filled canal connecting the ampulla pit with the surface of the skin. The organs are located intra-epithelially and are supported by a tiny dermal papilla. The cell elements of sensory epithelium are apically linked together by tight junctions. The free apical surface of the sensory cell bears several hundred densely packed stereocilia-like microvilli whereas the basal surface displays afferent neurosensory junctions with a pronounced round synaptic body. The compact uniform organization of the apical microvillous part shows a hexagonal pattern. A basal body was found in some sensory cells whereas a kinocilium was observed only in a single cell. The accessory cells have their free surface differentiated in a sparsely distributed and frequently-forked microvilli. The canal wall is built of two or three layers of tightly coalescent flat cells bordering on the lumen with branching microvilli. The ultrastructure of the content of the ampulla pit is presented. In the discussion stress is laid on the peculiarities of the natural history of Proteus anguinus that support the view that the morphologically-identified ampullary organs are electroreceptive. The structural characteristics of ampullary receptor cells are dealt with from the viewpoint of functional morphology and in the light of evolutionary hypotheses of ampullary organs.

Animals

Further freeze-fracture studies on the uterine epithelium of Salamandra salamandra (L.) (Amphibia, Urodela) using the antibiotic filipin.

The apical portion of the uterine lining of the ovoviviparous fire salamander, Salamandra salamandra, was studied by the freeze-fracture technique in conjunction with the polyene antibiotic filipin. Filipin-sterol complexes were found in the luminal plasmalemma and in the membranes limiting the mucous secretory granules typical of this epithelium. In all females, but particularly in non-pregnant females, more or less discrete clusters of filipin-sterol complexes were occasionally found overlying heavily affected secretory granules. The findings are discussed with regard to comparable results (Orci et al. 1980) based on the examination of collapsed and stretched urinary bladders of toads.

Animals

A Golgi study on the hypothalamus of amphibia. The neuronal typology.

The neuronal typology in the hypothalamus of the frog and the crested newt was studied by the Golgi technique. In the newt, piriform, multipolar or cerebrospinal fluid (CSF)-contacting neurons of relatively primitive type, according to the classification of Ramón-Moliner, are encountered in the preoptic area. Moreover, magnocellular neurons are impregnated. In the frog the preoptic area shows a more varied typology. The posterior hypothalami of the frog and the newt exhibit mainly bipolar CSF-contacting and piriform neurons. These latter are generally "tufted", but some bipolar of multipolar cells are encountered, especially in the frog. The simple anatomical organization of the amphibian hypothalamus corresponds well with the pattern of "generalized" integrative area where multimodal sensory inputs converge--including visceral information from cerebrospinal fluid by means of hypothalamic CSF-contacting sensors--to regulate the neuroendocrine outflow.

Animals

Retinal projections in the caecilian Ichthyophis kohtaoensis (Amphibia, Gymnophiona).

The retinal projections of the caecilian Ichthyophis kohtaoensis were investigated by anterograde transport of HRP. The optic tract forms two bundles in the diencephalon, a narrow medial bundle in the optic tectum, and a basal optic tract consisting of few fibres. Terminal fields are in the thalamus, pretectum, tectum, and as a circumscribed basal optic neuropile in the tegmentum. Thalamic, pretectal and tectal projections are contralateral as well as ipsilateral. The reduced but existing visual projection corresponds to a reduced but existing visually guided behaviour.

Amphibians

Comparative ultrastructural investigations of the uterine epithelium in the viviparous Salamandra atra Laur. and the ovoviviparous Salamandra salamandra (l.) (amphibia, urodela).

The uterine epithelium of the viviparous Salamandra atra and the ovoviviparous Salamandra salamandra was studied in non pregnant and ovulating females and in females during different stages of pregnancy. The epithelium of both species is organized in a monolayer. The epithelial cells are characterized by a moderate secretory activity, a variable amount of apical granules which include PAS-positive material and by some apical and basal exo- or endocytotic vesicles. Adjacent cells are joined by junctional complexes. The lateral surfaces form a tortuous boundary with adjoining cells which suggest that the epithelium is involved in transport. Sporadic light cells possess highly variable cytoplasmic inclusions and are not joined with neighbouring cells. Possibly they represent migratory cells. The entire epithelium, except for a small cranial portion of the uterus in S. atra, undergoes no remarkable morphological changes during the different physiological stages examined except that flattened cells seem to be more numerous in pregnant females. The results are discussed with regard to the possible supply of the developing young by the mother.

Animals

The vascularization of the neural stalk and the pars nervosa of the hypophysis in the toad, Bufo bufo (L.) (amphibia, anura). A comparative light microscopical and scanning electron microscopical study.

The angioarchitecture of the neural stalk and the encephaloposthypophysial portal system of the hypophysis of the toad, Bufo bufo (L.), was studied using three different methods. The neural stalk is mainly supplied by branches of the arteria infundibularis superficialis which form a widemeshed vascular network. Dorsally this network continues into the plexus of the pars nervosa. The vascularization of the pars nervosa is made up of the encephalo-posthypophysial portal system. This portal system consists of a hypothalamic branch (=portion), a mesencephalic and a mesencephalicbulbar branch (=portion). The hypothalamic branch was found to drain the pars ventralis of the tuber cinereum as well as more dorsal regions of the diencephalon. The mesencephalic-bulbar trunk enters the hypothalamic branch. The resulting common stem of the encephalo-posthypophysial portal vein the curves around the retroinfundibular communicating artery, crosses its ventral side and runs caudally. The secondary capillary plexus of the pars nervosa is characterized by well defined capillary plexus of the pars nervosa is characterized by well defined capillary networks which are located at the periphery of the parenchyma of the pars nervosa, thus forming a rostral, dorsal and ventro-caudal net. The central region of the parenchyma of the pars nervosa is supplied only by main branches of the encephalo-postpophysial portal vein. The venous drainage of the pars nervosa is via the vena hypophysea transversa.

Animals