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Morphological and fine structural features of the basilar papilla in ambystomatid salamanders (Amphibia; Caudata).

The morphology and fine structure of the basilar recess and basilar papilla were investigated in four species of salamanders from the family Ambystomatidae. The otic relationships of the recess and papilla to the proximal part of the lagena and saccule are described, and new terminology is suggested for the periotic relationships of the basilar recess to a diverticulum of an intracapsular periotic sac. The basilar papilla consists of supporting cells united laterally by gap junctions, capped by microvilli uniformly arranged around a short, central cilium, and hair cells that typically show several synapses with a single afferent nerve fiber, each marked by a rounded synaptic body surrounded by vesicles. In contrast to anuran basilar papillae, efferent nerve terminals were observed in synapse with hair cells and, rarely, upon afferent fibers. The distal half of the ambystomatid papilla contained hair cells capped by tall ciliary bundles, with kinocilia that show swellings near their tips with delicate attachments to adjacent tall stereocilia. A tectorial body covers only this region of the papilla. Hair cells with shorter stereocilia, situated in the proximal half and at the papillar margins, are related only to filamentous extensions of the tectorial body. The ambystomatid basilar recess and papilla are compared to auditory end-organs in other vertebrates, and it is suggested that a basic distinction can be made between aural neuroepithelia in amniotes versus that in nonamniotic vertebrate ears.

Ambystomatidae

Comparative features of the surface morphology of the basilar papilla in five families of salamanders (Amphibia; Caudata).

The surface morphology of the basilar recess and papilla was examined in 14 species of newts and salamanders selected from the five families of urodeles (Ambystomatidae, Salamandridae, Hynobiidae, Cryptobranchidae, and Amphiumidae) known to have this end-organ. In this sampling, the general organization of basilar structures is essentially similar across species investigated. The recess forms a tubular diverticulum of the proximal part of the lagena. One wall of the recess is associated with a diverticulum of the intracapsular periotic sac, and an adjacent wall is occupied by the basilar papilla. The papilla contained from as few as five hair cells in specimens of Taricha torosa to over 200 hair cells in Cryptobranchus allegheniensis. In most species, the papilla showed a morphological continuum between tall centrally or distally placed ciliary bundles and short ciliary bundles near the papillar margins. In certain species examined, tall bundles had kinocilia with swellings near their tips. Most forms showed a tendency to have groups of ciliary bundles morphologically polarized either toward or away from the saccule. In Cryptobranchus and Dicamptodon, many bundles had a random orientation. The gross and fine structural features of the basilar complex are compared in urodeles and anurans, and "generalized" features for the amphibian basilar complex are suggested. The basilar complex of Cryptobranchus is interpreted as being most generalized, representing a structural form from which most features of the basilar complex in other urodeles and anurans can be derived.

Ambystomatidae

Patterns of peripheral innervation of the tongue and hyobranchial apparatus in caecilians (Amphibia: Gymnophiona).

The innervation of the musculature of the tongue and the hyobranchial apparatus of caecilians has long been assumed to be simple and to exhibit little interspecific variation. A study of 14 genera representing all six families of caecilians demonstrates that general patterns of innervation by the trigeminal, facial, glossopharyngeal, and vagus nerves are similar across taxa but that the composition of the "hypoglossal" nerve is highly variable. Probably in all caecilians, spinal nerves 1 and 2 contribute to the hypoglossal. In addition, in certain taxa, an "occipital," the vagus, and/or spinal 3 appear to contribute fibers to the composition of the hypoglossal nerve. These patterns, the lengths of fusion of the contributing elements, and the branching patterns of the hypoglossal are assessed according to the currently accepted hypothesis of phylogenetic relationships of caecilians, and of amphibians. An hypothesis is proposed that limblessness and a simple tongue, with concomitant reduced complexity of innervation of muscles associated with limbs and the tongue, has released a constraint on pattern of innervation. As a consequence, a greater diversity and, in several taxa, greater complexity of neuroanatomical associations of nerve roots to form the hypoglossal are expressed.

Amphibians

Comparative anatomy and phylogeny of the cloacae of salamanders (Amphibia: Caudata). VI. Ambystomatidae and Dicamptodontidae.

Histology of the cloacae of Rhyacotriton olympicus and representative species from the genera Ambystoma and Dicamptodon was examined by light microscopy. Females of Ambystoma possess sperm storage glands, the spermathecae, as well as ventral glands and dorsal glands, both of uncertain function. Females of Ambystoma examined from the subgenus Linguaelapsus differ from those in the subgenus Ambystoma by possessing more extensive ventral gland clusters and a shorter cloacal tube. Females of Dicamptodon possess spermathecae and ventral glands, but differ in cloacal conformation from females of Ambystoma and lack the dorsal glands. Females of R. olympicus possess more extensive epidermal lining in the cloaca than that found in females of Ambystoma and Dicamptodon, and the only glands present are spermathecae, which cluster around a tube in the dorsal roof. Males of Ambystoma, Dicamptodon, and R. olympicus possess five types of cloacal glands (dorsal pelvic glands, lateral pelvic glands, anterior ventral glands, posterior ventral glands, and Kingsbury's glands) that function in spermatophore formation, and vent glands that may produce a courtship pheromone. In Ambystoma and Dicamptodon, vent glands secrete along the medial borders of the cloacal orifice. Males of A. opacum and A. talpoideum differ from males of other species examined from the two genera by possessing more extensive vent glands. Males of R. olympicus possess unique vent glands in which tubules secrete onto the surface of vent lobes lateral to the posterior end of the cloacal orifice, and distal ends of the glands pass anteriorly, superficial to the fascia enclosing the other cloacal glands. The results from analysis of cloacal anatomy support other data indicating that Ambystoma and Dicamptodon are sister groups, and that Rhyacotriton olympicus is not closely related to either of the other two genera and merits placement in a separate family.

Ambystomatidae

Glomerular development and growth of the renal blood vascular system in Xenopus laevis (Amphibia: Anura: Pipidae) during metamorphic climax.

Microcorrosion casts of the renal vascular system of tadpoles of the Clawed Frog, Xenopus laevis, were observed by scanning electron microscopy. Glomerular differentiation was studied qualitatively and quantitatively during developmental stages 56-66 (metamorphic climax). The general structure of the renal vascular system corresponds to the pattern commonly found in anurans; however, the arterial supply has conspicuous connecting vessels that supply groups of glomeruli. In the dorsal part of the kidney, qualitative differentiation of glomerular structures precedes quantitative growth. The ventral part of the kidney has larger, well-developed renal corpuscles of nearly adult appearance. Four developmental stages of glomerulogenesis are distinguished morphologically and their glomerular and vascular growth is analyzed.

Animals

Spermathecal cytology of Ambystoma opacum (Amphibia: Ambystomatidae) and the phylogeny of sperm storage organs in female salamanders.

Sperm storage glands, spermathecae, were examined from mated female Ambystoma opacum during the breeding season. No differences occur in the spermathecal ultrastructure of individuals sacrificed prior to oviposition and those sacrificed within 3 days of removal from tended clutches of recently oviposited eggs. The simple tubuloalveolar glands produce two types of secretory vacuoles. Apical secretory vacuoles contain glycosaminoglycans for export into the lumen to bathe stored sperm, perhaps providing the chemical/osmotic environment necessary for sperm quiescence. The other type of secretory vacuole contains an unsaturated lipid that is produced for export into the connective tissue surrounding the spermathecae. The role of this secretion may involve the contraction of myoepithelial cells, resulting in sperm expulsion. Some sperm undergo degradation in the spermathecal epithelium, and an interepithelial leukocyte was observed in one specimen. Apical secretory vacuoles and sperm are absent from the spermathecae of a specimen sacrificed 62 days after removal from a tended egg clutch. This is the first report on the spermathecal cytology of a salamander from the Ambystomatidase, and comparisons with salamanders from other families provide a morphological basis for considering spermathecae polyphyletic within the Caudata.

Ambystoma

Mechanical properties of the skin of Xenopus laevis (Anura, Amphibia).

The skin of the aquatic pipid frog, Xenopus laevis, was examined for specific biomechanical features: 1) thickness, 2) maximal strain at break (epsilon f), 3) tensile strength (sigma m), 4) modulus of elasticity (E, stiffness), and 5) the area under the stress-strain curve (W) (breaking energy, toughness). Skin freshly removed from dorsal, ventral, and lateral areas of the body was subjected to uniaxial tension. In both sexes, the dorsal skin is thicker than the ventral. The skin of male frogs was consistently thinner in all body regions than that of females. Most biomechanical parameters showed a considerable range of values in both males (epsilon f = 59-63%, sigma m = 15-16.5 MPa, E = 33.5-38.4 MPa, W = 3.8-4.5 MJ/m3) and females (epsilon f = 102-126%, sigma m = 11.5 MPa, E = 10.4-12 MPa, W = 5.2-6.7 MJ/m3). The disparate epsilon f values in males (low) and females (high) might reflect sexual dimorphism. Static stress-strain curves were typically J-shaped; with the exception of a "toe," the curves rose approximately linearly with increasing strain. The skin of X.laevis, although heterogeneous in structure, possesses features similar to those found in tissues with aligned collagen fibers such as tendons or fish skin. However, in anurans, the skin seems to play a more passive mechanical role during locomotion than in fish.

Animals

Cranial features of dendrobatid larvae (Amphibia: Anura: Dendrobatidae).

The larval neurocranium and visceral arches of seven dendrobatid species representing four genera are described, based on cleared-and-stained and serially sectioned specimens. A variety of characters is shared by all seven species. Larval features do not substantiate the assumption of close ranoid affinities of the Dendrobatidae. Instead dendrobatid larvae share features such as the special quadripartite cartilago suprarostralis, the lack of the larval processus oticus, the presence of three foramina acustica, and the lack of a foramen perilymphaticum accessorius with many bufonoid larvae. The first of these characters is unique to bufonids, hylids, dendrobatids, and some New World leptodactylids; the other characters also occur in pelobatids and are presumably plesiomorphic for the Neobatrachia. The free proximal ends of Ceratobranchialia II and III are an autapomorphy of the Dendrobatidae supporting the monophyly of the family. Some features of the cranium are paedomorphic: low cartilago orbitalis, lack of connection between cartilage orbitalis and otic capsule (most species), and vestigal taeniae tecti. New anatomical terms are introduced.

Animals

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

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