Effects of Thermal Stress on Hepatic Melanomacrophages of Eupemphix nattereri (Anura)

Melanomacrophages are the pigmented cells present in the hematopoietic organs. Besides melanin, hemosiderin and lipofuscin are also observed in the melanomacrophages. For the liver, however, numerous studies relate these cells to immunological and metabolic functions. Therefore, the aim of this study is to evaluate the hepatic metabolism by quantifying melanin, hemosiderin and lipofuscin in the anuran Eupemphix nattereri submitted to varying thermal conditions. E. nattereri adult males were separated into three groups, as follows: (i) five animals in the control group were kept at room temperature (27°C); (ii) 30 animals were submitted to hyperthermic (35.1°C); and (iii) 30 to hypothermic (18.9°) conditions. In each experiment, the animals were analyzed and separated into two different treatments: (1) immediately after undergoing the stress; and, (2) after recovering from the stress caused by the stimulus, at three distinct times (12 hr, 24 hr, and 48 hr). Both hyperthermia and hypothermia decreased hepatic pigmentation after thermal stress. The recovered animals of both experimental treatments showed as much pigmentation as the control animals. Thermal stress alters hemosiderin and lipofuscin as well, which may be related to liver function catabolism. In conclusion, liver pigmentation decreased due to temperature variation and duration of thermal stimulation to which the animals were exposed. The increase in temperature rather than hypothermia led to more drastic physiological disorders. In this study, we observed that thermal stress for a short period compromises the morphology and liver function, as observed by the changing pigmentation of melanomacrophages. These analyses can be used as biomarkers of environmental effects. Anat Rec, 297:864–875, 2014. © 2014 Wiley Periodicals, Inc.     

Cobaltic lysine study of the morphology and distribution of the cranial nerve efferent neurons (motoneurons and preganglionic parasympathetic neurons) and rostral spinal motoneurons in the Japanese toad

The morphology and distribution of the cranial nerve motoneurons (except III, IV, and VI) and rostral spinal motoneurons were systematically studied in the Japanese toad (Bufo japonicus) by retrograde labelling with cobaltic lysine complex. The cobaltic lysine clearly labelled whole neurons, i.e., cell bodies, proximal and distal dendrites, and axons. The branchial motoneurons (V, VII, IX, and X) had similar morphological characteristics and formed a more-or-less continuous cell column through the brainstem. The dendrites could be grouped mainly into the dorsomedial and the ventrolateral dendritic arrays. The dorsomedial dendrites formed a dendritic plexus in the subependymal gray matter, which extended as far peripherally as beneath the ependymal layer. The ventrolateral dendrites formed a broom-like dendritic plexus in the lateral to ventrolateral white matter. They usually extended as far peripherally as the pial surface. The rostrocaudal extent of the dendritic field was also wide and usually exceeded the motor nuclear boundaries. The hypoglossal motoneurons were grouped into the dorsomedial and ventrolateral cell groups, and the latter was considered to be part of the rostral spinal motoneuron column, from their morphology and distribution. The former had well-differentiated dendrites and occupied a more medial position than the branchial motoneurons. Besides the equivalent of the dorsomedial and ventrolateral dendritic arrays of the branchial motoneurons, they had dorsal and commissural dendrites. The accessory motoneurons had morphological characteristics and a distribution pattern similar to those of the rostral spinal motoneurons rather than the branchial motoneurons. The rostral spinal motoneurons had morphological characteristics somewhat different from the branchial motoneurons and the hypoglossal motoneurons (dorsomedial group). Functional implications of the motoneuron morphology are discussed, mainly based on the present results and earlier anatomical and physiological studies of the spinal motoneurons. The present study also revealed the anatomical features of the preganglionic parasympathetic neurons supplying some cranial nerves. These neurons had small somata with less elaborate dendrites and formed an almost continuous cell column that occupied a more dorsal position than the motoneurons of the corresponding nerve. They are thought to be homologous to the salivatory nucleus and the dorsal motor nucleus of the vagus. The basic anatomical organization of the general visceral efferent column seems to be similar throughout vertebrates.     

Invasion, stress, and spinal arthritis in cane toads

The impact of invasive species on biodiversity has attracted considerable study, but impacts of the invasion process on the invaders themselves remain less clear. Invading species encounter conditions different from those in their ancestral habitats and are subject to intense selection for rapid dispersal. The end result may be significant stress on individual organisms, with consequent health problems. Our studies on invasive cane toads in Australia reveal severe spinal arthritis in approximately 10% of large adult toads, associated with the same factors (large body size, frequent movement, and relatively long legs) that have enabled toads to invade so rapidly across the Australian tropics.     

Antimicrobial peptides from the skin of the Japanese mountain brown frog,Rana ornativentris

Abstract: Six peptides with antimicrobial activity were isolated from an extract of freeze‐dried skin of the Japanese mountain brown frog Rana ornativentris. Two structurally related peptides (brevinin‐20a GLFNVFKGALKTAGKHVAGSLLNQLKCKVSGGC, 11 nmol/g dried tissue, and brevinin‐20b GIFNVFKGALKTAGKHVAGSLLNQLKCKVSGEC, 170 nmol/g) belong to the brevinin‐2 family, previously identified in Asian and European, but not North American, Ranid frogs. Four peptides (temporin‐1Oa FLPLLASLFSRLL.NH₂, 13 nmol/g; temporin‐1Ob FLPLIGKILGTI L.NH₂, 350 nmol/g; temporin‐1Oc FLPLLASLFSRLF.NH₂, 14 nmol/g; and temporin‐1Od FLPLLASLFSGLF.NH₂, 8 nmol/g) are members of the temporin family first identified in the European common frog Rana temporaria but also found in the skins of North American Ranids. The brevinin‐2 peptides showed broad‐spectrum activity against the gram‐positive bacterium, Staphylococcus aureus, the gram‐negative bacterium, Escherichia coli and the yeast Candida albicans, whereas the temporins showed potent activity only against S. aureus. The brevinins and temporins belong to the class of cationic antimicrobial peptides that adopt an amphipathic α‐helical conformation but it is significant that temporin‐1Od, which lacks a basic amino acid residue, is still active against S. aureus (minimum inhibitory concentration=13 µm compared with 2 µm for temporin‐1Oa). This suggests that strong electrostatic interaction between the peptide and the negatively charged phospholipids of the cell membrane is not an absolute prerequisite for antimicrobial activity.     

Temperature Dependence of Anuran Distortion Product Otoacoustic Emissions

To study the possible involvement of energy-dependent mechanisms in the transduction of sound within the anuran ear, distortion product otoacoustic emissions (DPOAEs) were recorded in the northern leopard frog over a range of body temperatures. The effect of body temperature depended on the stimulus levels used and on the hearing organ under investigation. Low-level DPOAEs from the amphibian papilla (AP) were reversibly depressed for decreased body temperatures. Apparently, DPOAE generation in the AP depends on metabolic rate, indicating the involvement of active processes in the transduction of sound. In contrast, in the other hearing organ, the basilar papilla (BP), the effects of body temperature on DPOAEs were less pronounced, irrespective of the stimulus levels used. Apparently, metabolic rate is less influencing DPOAE generation. We interpret these results as evidence that no amplifier is involved in sound transduction in the BP. The passive functioning of the anuran BP would place this hearing organ in a unique position within tetrapod hearing, but may actually be beneficial to ectothermic species because it will provide the animal with a consistent spectral window, regardless of ambient or body temperature.     

Postmetamorphic changes in the lumbar lateral motor column in relation to muscle growth in the toad,Bufo americanus

Motoneuron number and size (nuclear cross-sectional area) were measured from serially sectioned spinal cords of Bufo americanus to investigate the relation between changes in the lumbar lateral motor column (L-LMC) and postmetamorphic increases in hindlimb muscle fiber number. Previous studies of neuron number in a variety of anuran species reported a correlation between number and body size, suggesting the possible addition of neurons during growth. Our results show a poor correlation between motoneu-ron number and body size with at most a 25% increase in neuron number occurring over the body size range where previous work had shown a hind-limb muscle fiber increase often to 20-fold. Thus, most new muscle fibers must be incorporated into motor units that exist at metamorphosis. Moto-neurons, but not ependymal cells, showed a significant size increase with increasing body size; this is perhaps related to an increased motor unit size that results from axonal sprouting. The range of variation of L-LMC cell numbers in newly metamorphosed toads was nearly equal to that of all other toads examined. This suggests that the weak correlations between motoneu-ron number and size observed in this and previous studies may be due to differential survival of individuals with larger neuron populations rather than to postmetamorphic addition of motoneurons. Our findings also show a strong bilateral correlation of motoneuron numbers, a finding suggesting that factors other than peripheral size may be important in regulating motoneuron number.     

Leaping lopsided: A review of the current hypotheses regarding etiologies of limb malformations in frogs

Recent progress in the investigation of limb malformations in free‐living frogs has underlined the wide range in the types of limb malformations and the apparent spatiotemporal clustering of their occurrence. Here, we review the current understanding of normal and abnormal vertebrate limb development and regeneration and discuss some of the molecular events that may bring about limb malformation. Consideration of the differences between limb development and regeneration in amphibians has led us to the hypothesis that some of the observed limb malformations come about through misdirected regeneration. We report the results of a pilot study that supports this hypothesis. In this study, the distal aspect of the right hindlimb buds of X. laevis tadpoles was amputated at the pre–foot paddle stage. The tadpoles were raised in water from a pond in Minnesota at which 7% of surveyed newly metamorphosed feral frogs had malformations. Six percent (6 of 100) of the right limbs of the tadpoles raised in pond water developed abnormally. One truncated right limb was the only malformation in the control group, which was raised in dechlorinated municipal water. All unamputated limbs developed normally in both groups. Three major factors under consideration for effecting the limb malformations are discussed. These factors include environmental chemicals (primarily agrichemicals), encysted larvae (metacercariae) of trematode parasites, and increased levels of ultraviolet light. Emphasis is placed on the necessary intersection of environmental stressors and developmental events to bring about the specific malformations that are observed in free‐living frog populations. Anat Rec (New Anat) 265:228–245, 2001. Published 2001 Wiley‐Liss, Inc.     

Lumbar lateral motor column development in triploid Xenopus laevis

The effects of increasing ploidy on the development of the lumbar lateral motor column (L-LMC) in Xenopus laevis were investigated in order to determine how early events contribute to producing the significant difference in the average number of motoneurons present in diploid and triploid animals after cell death (Sperry: J. Comp. Neurol. 277:499-508, '88). From naturally occurring diploid and experimentally produced triploid siblings at two stages prior to significant amounts of neuronal cell death, at one stage during the peak period of cell death, and at one stage after cell death, the L-LMC motoneurons were counted and nuclear cross-sectional areas were measured. At stages before and after cell death, the average nuclear cross-sectional areas of motoneurons and of other cells that were also measured were greater in the triploids, while the average number of motoneurons and motoneuron density (the mean number of cells per section) were less. Average body size and average motor column length in diploid and triploid animals were equal at each of the stages. The general characteristics of L-LMC development that have been widely noted in diploids, an increase in cell size accompanied by a decrease in cell number, were also observed in the triploid animals. However, not only were these general features present in the triploids, but the increase in average motoneuron size and the decrease in average motoneuron number in diploids and triploids were roughly equal when scaled to the general differences in nuclear size or to the difference in the average number of motoneurons present prior to cell death.(ABSTRACT TRUNCATED AT 250 WORDS)     

Skin of the Red Eye Tree Frog Agalychnis Callidryas (Hylidae,Phyllomedusinae) Contains Lipid Glands of the Type Described in the Genus Phyllomedusa

Several anuran species of the genus Phyllomedusa are known to possess specialized cutaneous glands producing lipids and exhibit a peculiar wiping behavior. This behavior is a stereotyped repertory of fore and hind limb movements distributing hydrophobic molecules onto the body surface and reducing evaporative water loss. No reports are presently available on the occurrence of lipid glands in other phyllomedusine genera, and data on the structure of the secretory units specialized for the production of cutaneous lipids are still unclear. The present report is aimed to answer both questions: it describes lipid glands of the Phyllomedusa type in Agalychnis callidryas and provides light and transmission electron microscope evidence of the syncytial structure of their secretory units, a typical feature of serous glands in anuran skin. This morphological trait supports the hypothesis that lipid glands are a specialized subset of the anuran serous glands, and underlines their flexible role in the skin adaption to sub‐aerial environments. Anat Rec, 300:503–506, 2017. © 2016 Wiley Periodicals, Inc.     

Sesamoids in Anurans: New Data,Old Issues

Sesamoids are skeletal elements rarely considered in studies of the vertebrate skeleton. In this work, we integrate ontogenetic data of anuran sesamoids in two species (Leptodactylus latinasus and Pleurodema cf. guayapae), the related structures (tendons, muscles, and joints) in L. latinasus, and a survey of sesamoid distribution in 185 anuran taxa. Our main goals are: (1) to contribute to the knowledge of the comparative anatomy of sesamoids in tetrapods; (2) to provide additional developmental evidence to interpret the ontogenetic pattern of sesamoids in anurans, as a key to elucidate that of tetrapods in general; (3) to provide data about tendon development in relation to sesamoid development in anurans for the first time; and (4) to propose a pattern of anuran sesamoid distribution. The homologies of sesamoids across tetrapods are discussed here. Observations were made in cleared and stained skeletal whole‐mounts. Fifty‐four sesamoids were found in anurans, thirty‐seven of which occur in L. latinasus. The traditional point of view of embedded sesamoids always resulting from biomechanical stimuli of a previously existing tendon is not sustained by our data. Many sesamoids arise before the differentiation of a tendinous tissue. Our survey results in a data set where the two big anuran clades, Hyloides (12 families) and Ranoides (14 families), were represented. The matrix has 38% missing entries. Most of the surveyed sesamoids have multiple origins, with only three of them (about 19%) having one origin. Anat Rec 293:1646–1668, 2010. © 2010 Wiley‐Liss, Inc.