Diversity of cortical bone morphology in anuran amphibians

The cortical bones of mammals, birds, and reptiles are composed of a complex of woven bone and lamellar bone (fibrolamellar bone) organized into a variety of different patterns; however, it remains unclear whether amphibians possess similar structures. Importantly, to understand the evolutionary process of limb bones in tetrapods, it is necessary to compare the bone structure of amphibians (aquatic to terrestrial) with that of amniotes (mostly terrestrial). Therefore, this study compared the cortical bones in the long bones of several frog species before and after metamorphosis. Using micro-computed tomography (CT), we found that the cortical bones in the fibrolamellar bone of Xenopus tropicalis (Pipoidea superfamily) and Lithobates catesbeianus (Ranoidea superfamily) froglets are dense, whereas those of Ceratophrys cranwelli (Hyloidea superfamily) are porous. To clarify whether these features are common to their superfamily or sister group, four other frog species were examined. Histochemical analyses revealed porous cortical bones in C. ornata and Lepidobatrachus laevis (belonging to the same family, Ceratophryidae, as C. cranwelli). However, the cortical bones of Dryophytes japonicus (Hylidae, a sister group of Ceratophryidae in the Hyloidea superfamily), Microhyla okinavensis (Microhylidae, independent of the Hyloidea superfamily), and Pleurodeles waltl, a newt as an outgroup of anurans, are dense with no observed cavities. Our findings demonstrate that at least three members of the Ceratophryidae family have porous cortical bones similar to those of reptiles, birds, and mammals, suggesting that the process of fibrolamellar bone formation arose evolutionarily in amphibians and is conserved in the common ancestor of amniotes.     

Neuromast topography in anuran amphibians

Generalized anuran tadpoles across families exhibit a similar neuromast morphology on their heads, as follows: (1) all neuromast lines known for anurans are present; (2) within these lines total neuromast number ranges from about 250 to 320; (3) neuromasts form linear stitches composed of two to three, but sometimes up to five, neuromasts; (4) neuromast linear dimensions are ? 10 μm; and (5) neuromasts contain ? 15 hair cells. Compared with generalized forms, stream, arboreal, carnivorous, and desert-pond forms have fewer neuromasts but they contain more hair cells. They do not, however, form stitches. Obligate midwater suspension-feeding forms, including Xenopus (Pipidae), Rhinophrynus (Rhinophyrnidae), and Phrynomerus (Microhylidae), form stitches that contain > six, but potentially up to 18 or more, loosely aggregated neuromasts. Xenopus and Rhinophrynus have large neuromasts (up to 40 μm across). Chiasmocleis (Microhylidae) tadpoles form stiches that are linearly arranged with up to ten neuromasts. Whereas urodeles can have more than one neuromast row per line and may form both linear and transverse stitches, anurans have only one row of neuromasts per line and form only transverse stitches. Neuromasts in anurans tend to be smaller and more circular than in urodeles and positioned flush with the epidermal surface. A greater percentage of anurans form stitches, and anurans have greater intrafamilial variation in stitch formation than do urodeles.     

Brain regeneration in anuran amphibians

Urodele amphibians are highly regenerative animals. After partial removal of the brain in urodeles, ependymal cells around the wound surface proliferate, differentiate into neurons and glias and finally regenerate the lost tissue. In contrast to urodeles, this type of brain regeneration is restricted only to the larval stages in anuran amphibians (frogs). In adult frogs, whereas ependymal cells proliferate in response to brain injury, they cannot migrate and close the wound surface, resulting in the failure of regeneration. Therefore frogs, in particular Xenopus, provide us with at least two modes to study brain regeneration. One is to study normal regeneration by using regenerative larvae. In this type of study, the requirement of reconnection between a regenerating brain and sensory neurons was demonstrated. Functional restoration of a regenerated telencephalon was also easily evaluated because Xenopus shows simple responses to the stimulus of a food odor. The other mode is to compare regenerative larvae and non-regenerative adults. By using this mode, it is suggested that there are regeneration-competent cells even in the non-regenerative adult brain, and that immobility of those cells might cause the failure of regeneration. Here we review studies that have led to these conclusions.     

Control of breathing in anuran amphibians

The primary role of the respiratory system is to ensure adequate tissue oxygenation, eliminate carbon dioxide and help to regulate acid-base status. To maintain this homeostasis, amphibians possess an array of receptors located at peripheral and central chemoreceptive sites that sense respiration-related variables in both internal and external environments. As in mammals, input from these receptors is integrated at central rhythmogenic and pattern-forming elements in the medulla in a manner that meets the demands determined by the environment within the constraints of the behavior and breathing pattern of the animal. Also as in mammals, while outputs from areas in the midbrain may modulate respiration directly, they do not play a significant role in the production of the normal respiratory rhythm. However, despite these similarities, the breathing patterns of the two classes are different: mammals maintain homeostasis of arterial blood gases through rhythmic and continuous breathing, whereas amphibians display an intermittent pattern of aerial respiration. While the latter is also often rhythmic, it allows a degree of fluctuation in key respiratory variables that has led some to suggest that control is not as tight in these animals. In this review we will focus specifically on recent advances in studies of the control of ventilation in anuran amphibians. This is the group of amphibians that has attracted the most recent attention from respiratory physiologists.     

Sound source perception in anuran amphibians

Sound source perception refers to the auditory system's ability to parse incoming sensory information into coherent representations of distinct sound sources in the environment. Such abilities are no doubt key to successful communication in many taxa, but we know little about their function in animal communication systems. For anuran amphibians (frogs and toads), social and reproductive behaviors depend on a listener's ability to hear and identify sound signals amid high levels of background noise in acoustically cluttered environments. Recent neuroethological studies are revealing how frogs parse these complex acoustic scenes to identify individual calls in noisy breeding choruses. Current evidence highlights some interesting similarities and differences in how the auditory systems of frogs and other vertebrates (most notably birds and mammals) perform auditory scene analysis.     

Evolutionary transformations of ontogenesis in anuran amphibians

A review of the recent published data on ontogenesis of direct developing and marsupial frogs. The development of these representatives of anuran amphibians seems to be evolutionary advanced and considerably differs from the development of species traditionally used in amphibian embryology.     

Hormones and acoustic communication in anuran amphibians

Circulating hormone levels can mediate changes in the quality of courtship signals by males and/or mate choice by females and may thus play an important role in the evolution of courtship signals. Costs associated with shifts in hormone levels of males, for example, could effectively stabilize directional selection by females on male signals. Alternatively, if hormone levels affect the selection of mates by females, then variation in hormone levels among females could contribute to the maintenance of variability in the quality of males' signals. Here, I review what is known regarding the effects of hormone levels on the quality of acoustic signals produced by males and on the choice of mates by females in anuran amphibians. Surprisingly, despite the long history of anuran amphibians as model organisms for studying acoustic communication and physiology, we know very little about how variation in circulating hormone levels contributes to variation in the vocal quality of males. Proposed relationships between androgen levels and vocal quality depicted in recent models, for example, are subject to the same criticisms raised for similar models proposed in relation to birds, namely that the evidence for graded effects of androgens on vocal performance is often weak or not rigorously tested and responses seen in one species are often not observed in other species. Although several studies offer intriguing support for graded effects of hormones on calling behavior, additional comparative studies will be required to understand these relationships. Recent studies indicate that hormones may also mediate changes in anuran females' choice of mates, suggesting that the hormone levels of females can influence the evolution of males' mating signals. No studies to date have concurrently addressed the potential complexity of hormone-behavior relationships from the perspective of sender as well as receiver, nor have any studies addressed the costs that are potentially associated with changes in circulating hormone levels in anurans (i.e., life-history tradeoffs associated with elevations in circulating androgens in males). The mechanisms involved in hormonally induced changes in signal production and selectivity also require further investigation. Anuran amphibians are, in many ways, conducive to investigating such questions.     

Retina and lens regeneration in anuran amphibians

Anuran amphibians can regenerate the retina through differentiation of stem cells in the ciliary marginal zone and through transdifferentiation of the retinal pigmented epithelium. By contrast, the regeneration of the lens has been demonstrated only in larvae of species belonging to the Xenopus genus, where the lens regenerates through transdifferentiation of the outer cornea. Retinal pigmented epithelium to neural retina and outer cornea to lens transdifferentiation processes are triggered and sustained by signaling molecules belonging to the family of the fibroblast growth factor. Both during retina and lens regeneration there is a re-activation of many of the genes which are activated during development of the eye, even though the spatial and temporal pattern of gene expression is not a simple repetition of that found in development.     

Morphometric relationships of take-off speed in anuran amphibians

Locomotory speed correlates with muscle mass (determining force and stride rate), limb length (stride rate and distance), and laterally compressed body trunk (force and stride distance). To delineate generalization of the locomotory-morphometric relationships specifically in anuran amphibians, we investigated take-off speed and the three morphological variables from seven species, Rana nigromaculata, R. rugosa, and Bombina orientalis, Eleuthrodectilus fitzingeri, E. diastema, Bufo typhonius, Colostethus flotator and Physalaemus pustulosus. The fastest jumper E. fitzingeri (3.41 m s(-1)) showed 2.49-fold greater speed than the slowest B. typhonius. Take-off speed correlated well with both thigh muscle mass relative to body mass and hindlimb length relative to snout-vent length (HL/SVL), but poorly correlated with the inter-ilial width relative to SVL. The best morphological predictor was HL/SVL (speed=-3.28+3.916 HL/SVL, r=0.968, P<0.0001), suggesting that anuran take-off speed is portrayed well with high gear and acceleration distance characterized by hindlimbs.     

Developmental changes in interrenal responsiveness in anuran amphibians

Basal activity of the hypothalamo-pituitary-interrenal (HPI)axis changes over development in larval amphibians, but developmentof the responsiveness of this axis to an external stressor hasnot been studied. We compared developmental changes in whole-bodycorticosterone content of two anuran amphibian species, Ranapipiens (family Ranidae) and Xenopus laevis (family Pipidae).We also examined developmental changes in the responsivenessof the HPI axis by subjecting tadpoles of different developmentalstages to a laboratory shaking/confinement stress and to ACTHinjection. We measured whole-body corticosterone content asan indicator of the activity of the HPI axis. Whole-body corticosteronecontent of R. pipiens remained low during premetamorphosis andprometamorphosis but increased dramatically at metamorphic climaxand remained elevated in juvenile frogs. By contrast, whole-bodycorticosterone content of X. laevis was highest during premetamorphosis,declined at the onset of prometamorphosis, increased at metamorphicclimax and remained at climax levels in juvenile frogs. Premetamorphicand prometamorphic tadpoles of both species showed strong corticosteroneresponses to both shaking stress and ACTH injection. The magnitudeand pattern of response differed among developmental stages,with premetamorphic tadpoles of both species showing greaterresponsiveness to stress and ACTH. Our results show that interrenalresponsiveness is developed in premetamorphic tadpoles, suggestingthat at these stages tadpoles are capable of mounting an increasein stress hormone production in response to changes in the externalenvironment. Our results also highlight the importance of comparativestudies in understanding the development of the stress axis.     

Patterns of carpal development among anuran amphibians

The unity and diversity of developmental processes in the vertebrate limb have singular importance in the interpretation of evolutionary hypotheses of tetrapod diversification. In anurans, the intraordinal diversity of forelimbs seems to be related to the fusion of distal carpals, whereas proximal carpals are invariable. However, there are different ontogenetic pathways involved in the differentiation of proximal carpals. This study presents a comparative analysis of early developmental features in one archeobatrachian and 23 neobatrachian species representing five families and explores the variability in the differentiation of carpal cartilages. We found new evidence supporting the presence of an embryonic intermedium that incorporates with the ulnare. Difference between the pipid Xenopus and the neobatrachians is interpreted as a change in the rate of differentiation of Distal Carpal 5 that does not affect the developmental pattern of digits. The developmental variability exhibited by the intermedium, radiale, and Element Y is combined in patterns that converge on the same adult carpal morphology among neobatrachians; these patterns appear to contain potentially useful phylogenetic information.     

Role of midbrain in the control of breathing in anuran amphibians

The present study was designed to explore systematically the midbrain of unanesthetized, decerebrate anuran amphibians (bullfrogs), using chemical and electrical stimulation and midbrain transections to identify sites capable of exciting and inhibiting breathing. Ventilation was measured as fictive motor output from the mandibular branch of the trigeminal nerve and the laryngeal branch of the vagus nerve. The results of our transection studies suggest that, under resting conditions, the net effect of inputs from sites within the rostral half of the midbrain is to increase fictive breathing frequency, whereas inputs from sites within the caudal half of the midbrain have no net effect on fictive breathing frequency but appear to act on the medullary central rhythm generator to produce episodic breathing. The results of our stimulation experiments indicate that the principal sites in the midbrain that are capable of exciting or inhibiting the fictive frequency of lung ventilation, and potentially clustering breaths into episodes, appear to be those primarily involved in visual and auditory integration, motor functions, and attentional state.     

Thermal dependence of behavioural performance of anuran amphibians

The thermal dependence of performance capacity was assessed in two anuran amphibians: Bufo boreas (western toad) and Rana pipiens (leopard frog). Quantitative measurements of performance showed that Bufo could sustain slow rates of walking for 10 min and cover greater distances than Rana, which initially jumped more vigorously but fatigued within 5 min. Changes in performance with changes in body temperature were virtually instantaneous, and performance exhibited no acclimation over 7 days. Within the range of temperatures studied, performance capacity increased with increasing body temperature and reached a maximum at 28 C in Bufo and 20 to 29 C in Rana. Performance capacity and the underlying metabolic processes had a similar thermal dependence within a species. The behavioural capacity for activity is apparently maximal for both species at body temperatures normally encountered in the field. Anuran behaviours requiring sustained activity (migration to breeding sites, mating, foraging) must therefore be markedly temperature-sensitive.     

Transdifferentiation of eye tissues in anuran amphibians: Analysis of the transdifferentiation capacity of the iris of Xenopus laevis larvae

Abstract. Lensectomized Xenopus laevis larvae are capable of regenerating a lens from the cells of the outer cornea. Unlike the outer cornea, the iris of larval Xenopus exhibits a high degree of phenotypic stability, even when it has been damaged to various degrees in order to stimulate its latent transdifferentiative competence. However, when isolated from its surrounding tissues and implanted in an appropriate site, the dorsal iris of larval Xenopus is capable of following a differentiative pathway different to that normally followed in situ. Our results show that, when such an implant is placed in the vitreous chamber of a lensectomized eye, the pigmented epithelial cells of the iris transdifferentiate into neural retina regardless of whether the iris stroma is present or not. Unlike the vitreous chamber, the environment of the anterior chamber of a lensectomized eye does not promote the transdifferentiative process of the iris. We suggest the existence of eye factors that promote retina-forming transformation of the iris and that are distributed in a gradient in lensectomized eyes.     

Evolutionary causes and consequences of sequential polyandry in anuran amphibians

Among anuran amphibians (frogs and toads), there are two types of polyandry: simultaneous polyandry, where sperm from multiple males compete to fertilize eggs, and sequential polyandry, where eggs from a single female are fertilized by multiple males in a series of temporally separate mating events, and sperm competition is absent. Here we review the occurrence of sequential polyandry in anuran amphibians, outline theoretical explanations for the evolution of this mating system and discuss potential evolutionary implications. Sequential polyandry has been reported in a limited number of anurans, but its widespread taxonomic and geographic distribution suggests it may be common. There have been no empirical studies that have explicitly investigated the evolutionary consequences of sequential polyandry in anurans, but species with this mating pattern share an array of behavioural, morphological and physiological characteristics, suggesting that there has been common sexual selection on their reproductive system. Sequential polyandry may have a number of adaptive benefits, including spreading the risk of brood failure in unpredictable environments, insuring against male infertility, or providing genetic benefits, either through good genes, intrinsic compatibility or genetic diversity effects. Anurans with sequential polyandry provide untapped opportunities for innovative research approaches that will contribute significantly to understanding anuran evolution and also, more broadly, to the development of sexual‐selection and life‐history theory.     

Control and interaction of the cardiovascular and respiratory systems in anuran amphibians

In anuran amphibians, respiratory rhythm is generated within the central nervous system (CNS) and is modulated by chemo- and mechanoreceptors located in the vascular system and within the CNS. The site for central respiratory rhythmogenesis and the role of various neurotransmitters and neuromodulators is described. Ventilatory air flow is generated by a positive pressure, buccal force pump driven by efferent motor output from cranial nerves. The vagus (cranial nerve X) also controls heart rate and pulmocutaneous arterial resistance that, in turn, affect cardiac shunts within the undivided anuran ventricle; however, little is known about the control of central vagal motor outflow to the heart and pulmocutaneous artery. Anatomical evidence indicates a close proximity of the centers responsible for respiratory rhythmogenesis and the vagal motoneurons involved in cardiovascular regulation. Furthermore, anurans in which phasic feedback from chemo- and mechanoreceptors is prevented by artificial ventilation exhibit cardiorespiratory interactions that appear similar to those of conscious animals. These observations indicate interactions between respiratory and cardiovascular centers within the CNS. Thus, like mammals and other air-breathing vertebrates, the cardio-respiratory interactions in anurans result from both feedback and feed-forward mechanisms.