Termination of the afferent fiber in the spinal cord of the turtle Testudo horsfieldi and three-dimensional reconstruction of the sensory-motoneuron connection

Horseradish peroxidase (HRP) tracing methods and subsequent computer reconstruction were used to study the structural organization of sensory-motoneuron connections in turtles. HRP was applied through suction electrodes to thin dorsal and ventral root filaments of superfused isolated lumbar spinal cord of the turtle Testudo horsfieldi. Single motoneurons were labeled ionophoretically with intracellular glass microelectrodes. Labeled elements were examined under a light microscope. The Eutectic neuron tracing system and its associated program were used for three-dimensional reconstructions and morphometry. The distribution of afferent fibers of the dorsal root and their terminations were presented in a new scheme in which new zones, in addition to those that were already well known, were shown, including the following: in the Lissauer zone, motor nuclei, and ventrolateral funiculus, as well as in the contralateral medial gray matter (laminae IV–V). Unlike in frogs, the motoneuron dendritic field in turtles was restricted to an ellipsoid space with a short axis in the rostrocaudal direction (300–500 µm). The afferent fibers of the dorsal root connected to motoneurons produced very short branches in a restricted rostrocaudal direction (50–70 μm). One fiber collateral of the dorsal root had about 80 synapse-like enlargements (approximately tenfold fewer than in frogs). Putative sensory-motoneuron contacts were found on the I–VII-order dendritic segments of the dorsal and ventro-medial dendritic trees. It was shown that, in turtles, only one first-order collateral of the dorsal root fiber participated in the sensory-motoneuron connection with a small number (about 4) of putative contacts, which is also one order less than in frogs. It is likely that the simplification of the synapse structure in turtles is compensated by a higher efficiency of the signal transmission comparable to that in mammals.     

Photoreceptor layer composition in the retina of the frog (Rana esculenta)

Retinas of Rana esculenta frogs were studied by light and electron microscopy in order to establish the photoreceptor layer composition. We found 56% red rods, 9% green rods, 19% single cones, and 16% double cones. This work provides the morphological basis for electrophysiological investigations concerning the mass receptor potential of isolated Rana esculenta retinas.     

Electroencephalographic dynamics of individual alpha peak frequency during the female ovarian-menstrual cycle

Electroencephalographic brain dynamics during the menstrual, follicular, preovulatory, ovulatory, luteal and premenstrual phases of the female ovarian-menstrual cycle were analyzed. All subjects were divided into 3 groups according to individual alpha peak frequency and reaction to rhythmical photostimulation. Data obtained confirm the necessity to carry out experimental and clinical psychophysiological studies of women in the phases of the ovarian-menstrual cycle.     

Melanomacrophage Aggregations and Their Age Relationships in the Yellow Mud Turtle,Kinosternon flavescens (Kinosternidae)

Microscopic examination of the pigment cell aggregations in the liver and spleen of mud turtles (Kinosternon flavescens) suggests that the morphology and behavior of these cells is consistent with the melanomacrophages described in teleost fishes and the pigmented “Kupffer cells” described in frogs and reptiles. These cells contain massive amounts of melanin, substantial lipofuscin, and some hemosiderin consistent with their phagocytic function. Similar-appearing isolated pigmented macrophages are solitary in the liver, spleen, lung, and kidney. Number and size of the largest hepatic aggregations increase almost linearly with turtle age so that in old turtles they may constitute up to 20% of the liver volume. This increase may result from hepatic recruitment of macrophages throughout the life of the turtle and suggests that size and number of melanomacrophage aggregations may serve as a marker for senescence in otherwise healthy turtles of this species.     

Edwardsiella tarda in freshwater catfish and their environment

Edwardsiella tarda was isolated from 47, 88, and 79% of skin, visceral, and dressed-fish samples, respectively. This species was also isolated from 30% of imported dressed fish, 75% of catfish pond water samples, 64% of catfish pond mud samples, and 100% of frogs, turtles, and crayfish from catfish ponds. The incidence of Edwardsiella increased during the summer months, as water temperatures increased. Of several isolation media evaluated, the most effective was selective enrichment in double-strength Salmonella-Shigella broth and subsequent plating on single-strength Samonella-Shigella agar. The significance of the incidence of Edwardsiella in catfish, catfish disease, and public health could not be substantiated.     

Edwardsiella tarda in freshwater catfish and their environment.

Edwardsiella tarda was isolated from 47, 88, and 79% of skin, visceral, and dressed-fish samples, respectively. This species was also isolated from 30% of imported dressed fish, 75% of catfish pond water samples, 64% of catfish pond mud samples, and 100% of frogs, turtles, and crayfish from catfish ponds. The incidence of Edwardsiella increased during the summer months, as water temperatures increased. Of several isolation media evaluated, the most effective was selective enrichment in double-strength Salmonella-Shigella broth and subsequent plating on single-strength Samonella-Shigella agar. The significance of the incidence of Edwardsiella in catfish, catfish disease, and public health could not be substantiated.     

Poison frogs rely on experience to find the way home in the rainforest

Among vertebrates, comparable spatial learning abilities have been found in birds, mammals, turtles and fishes, but virtually nothing is known about such abilities in amphibians. Overall, amphibians are the most sedentary vertebrates, but poison frogs (Dendrobatidae) routinely shuttle tadpoles from terrestrial territories to dispersed aquatic deposition sites. We hypothesize that dendrobatid frogs rely on learning for flexible navigation. We tested the role of experience with the local cues for poison frog way-finding by (i) experimentally displacing territorial males of Allobates femoralis over several hundred metres, (ii) using a harmonic direction finder with miniature transponders to track these small frogs, and (iii) using a natural river barrier to separate the translocated frogs from any familiar landmarks. We found that homeward orientation was disrupted by the translocation to the unfamiliar area but frogs translocated over similar distances in their local area showed significant homeward orientation and returned to their territories via a direct path. We suggest that poison frogs rely on spatial learning for way-finding in their local area.     

Freeze-fracture evidence for the presence of cholesterol in particle- free patches of basal disks and the plasma membrane of retinal rod outer segments of mice and frogs

The freeze-fracture technique was used to examine the membranes of the photoreceptors of mice and frogs. Particle-free patches were found in the plasma membrane and basal disk membranes of the outer segments of both mice and frogs housed at room temperature, but not in frogs kept in a cold room. These patches were shown not to be artifacts of cryoprotection or fixation, and they persisted when fresh isolated outer segments were frozen by an ultrarapid method. They were also found to persist in mouse rods when retinas were incubated and subsequently fixed at temperatures up to 80 degrees C. Cholesterol was implicated as a significant component of the patches by the observation that, in the outer segments, pits, induced by treatment with the sterol-specific polyene antibiotic filipin, were present in and confined to the particle-free patches. That these lesions are not inherently limited to particle-free membrane areas was evident in the apical plasma membrane of the photoreceptor inner segments, where particles and pits were intermixed. Treatment with saponin, a surface-active agent which specifically complexes cholesterol, resulted in the disappearance of the particle-free patches. Patches were found in basal disks of both mouse and frog rods but not in older disks nearer the pigment epithelium, which indicates that changes occur in the composition of disk membranes and/or in the molecular ordering of their protein and lipid components during the early phase of their transit from the base towards the apex of the outer segment.     

Catabolism of myo-Inositol to Precursors Utilized for De Novo Glycerolipid Biosynthesis

Systemic injection of [2-3H]myo-inositol into frogs resulted in the incorporation of more than half of the label into glycerolipid classes other than phosphoinositides in retinal rod outer segment membranes. Following methanolysis and differential extraction of isolated lipid classes, radioactivity was recovered primarily in the aqueous phase. After phospholipase C hydrolysis of the total membrane lipids, 97% of the radioactivity was extractable with organic solvents, and 70% of the label in lipids was in 1,2-diglycerides. These results indicate that the label was incorporated primarily into the glyceryl moiety of the membrane glycerolipids. Intraocular injection of frog eyes or in vitro incubation of frog retinas with [2-3H]myo-inositol resulted in the incorporation of radioactivity almost exclusively into phosphoinositides in rod outer segment membranes. Incubation of retinas with [U-14C]glucuronic acid did not result in the formation of labeled retinal lipids. These results suggest that myo-inositol can be catabolized systemically to precursors utilized for glycerolipid biosynthesis in the retina.     

Frogs and turtles: different ectotherm overwintering strategies

The ability of frogs and turtles to overwinter and to survive hypoxia and anoxia has long been a topic of interest. While data remains scant, the emerging picture shows fundamentally different approaches to overwintering in these two groups of ectotherms. Frogs are far more limited by availability of oxygen than are turtles, even at near-freezing ambient temperatures. The reasons for this probably involve the vastly greater cutaneous permeability of the former. With their extreme tolerance of anoxia and profound suppression of metabolism, overwintering in turtles should not be viewed as simply prolonged diving but rather as ectotherm hibernation. Their incredible diving capabilities are merely a spin-off of a successful overwintering strategy. The following discussion reviews the major physiological mechanisms involved in the overwintering strategies of these two ectotherm groups.     

The skull of Cteniogenys, a choristodere (Reptilia: Archosauromorpha) from the Middle Jurassic of Oxfordshire

The genus Ctemogenys was originally described as a lizard on the basis of isolated jaw fragments from the Upper Jurassic deposits of Como Bluff, Wyoming. The discovery of new material from a Middle Jurassic locality at Kirtlington, Oxfordshire, showed that Cteniogenys was not a lizard, but an early choristodere. The skull is represented by a collection of isolated bones, associated on the basis of fit and dermal sculpture pattern. The bones are here described and compared with those of the Late Cretaceous and Palaeocene choristoderes Champsosaurus and Simoedosaurus. Allowing for the much smaller size of the Middle Jurassic form, there is a close correspondence between the isolated bones of the three genera. Comparison with known choristoderes, based on an analysis of 53 derived character states, suggests that Ctemogenys is the most primitive of known genera. In general, the character states shared by all choristoderes support recent analyses which conclude that choristoderes arc derived from archosauromorph diapsids, not lepidosauromorphs as once thought. At Kirtlington, Cteniogenys forms part of a diverse microvertebrate assemblage including amiids, sharks, frogs, salamanders, lizards, mammals, crocodiles, pterosaurs, turtles and small dinosaurs.     

Antipredator Behavioral Responses of Native and Exotic Tadpoles to Novel Predator

Factors related to the invasion process, such as high abundance of invaders, residence time, and functional distinctiveness, are well documented, but less attention has been given to the effects of antipredator strategy of invasive species during colonization. In this study, we explored the antipredator strategy of an introduced species by comparing the predator avoidance behaviors of two native anuran species and one introduced("exotic") species in the presence of different predators. The two native anuran species used in the study were Black-spotted Pond Frog Rana nigromaculata and Terrestrial Frog Rana limnocharis. The introduced(invasive) species used was American bullfrog Lithobates catesbeianus. Chinese pond turtle Chinemys reevesii, Red-backed rat snake Elaphe rufodorsata, and Big-headed turtle Platysternon megacephalum were used as predator species. Chinese pond turtles and Red-backed rat snakes are native predators of Black-spotted Pond Frogs and Terrestrial Frogs, while Big-headed turtles are novel("unfamiliar") to the two frogs. All three predator species are novel("unfamiliar") to the American bullfrog. The results show that tadpoles of the two native species displayed behaviors of recognizing the two native predators, but did not display the capability of identifying the novel predator. Results from our study also suggest that American bullfrog tadpoles exhibited strong antipredator behavioral responses by displaying the capability of identifying "unfamiliar" predators without cohabitation history and prior exposure to them. Such antipredator behavioral responses could have resulted in more favorable outcomes for an invading species during the invasive introductory process.     

Comparative responses of microscopic fungi to ionizing radiation and light

Microscopic fungi of 47 species and 24 genera isolated from radioactive soil and other radioactive substrates of a 10-km alienation zone from Chernobyl Nuclear Power Plant were investigated. Only 19 % of such fungi showed toward the ionizing irradiation source positive radiotropism. Strains with best-defended radiotropism responded to the influence of weak white light. Both of these responses (radiotropism and photostimulation) were of an adaptive nature, were found in both dark-colored and light-colored micromycetes and were probably a strain-related feature.     

Dynamic characteristics of the human resonance EEG responses to rhythmic photostimulation

The development of the resonance EEG responses of the left and right occipital areas was studied in right-handed men during prolonged (12 or 120 s) rhythmic, photostimulation with the intensity of 0.7 J and frequencies of 6, 10, and 16 Hz. Analysis of the EEG fine spectral structure was applied to compare the accumulated baseline EEG spectra and EEG spectra during photostimulation, to observe the dynamics of the short-term spectra and to detect power changes in the EEG narrow spectral band sharply coincident with the stimulation frequency. The more pronounced EEG responses to photostimulation were observed in subjects with the initially low EEG baseline, α-rhythm. Two-minute flash trains produced a substantial increase in the EEG power within the stimulation frequency with superposed oscillatory processes with different periods. These fluctuations are considered a reflection of intricate interaction between the adaptive and resonance EEG responses to the presented intermittent stimulation. Under 12-s stimulation the resonance EEG responses are steadily recorded within the first 3 s of stimulation and immediately after the flash cessation EEG power at the stimulation frequency returns to the initial level. The resonance EEG responses were more pronounced in the right hemisphere than in the left one, especially, at the stimulation frequencies of 6 and 16 Hz. With increasing the stimulation frequency, the maximum of resonance EEG responses was reached earlier. Under the stimulation frequency of 6 Hz, the maximal response was recorded 9–12 s after the beginning of flashes, at the frequencies of 10 and 16 Hz, it was recorded within 3–6 and 3 s, respectively.     

Comparative effects of diverse vertebrate gonadotropins on androgen formation in vitro from testes of roosters and mice

A study was carried out to compare the androgen formation activity of gonadotropins from diverse vertebrate species by rooster and mouse testes in vitro. The dispersed testicular interstitial cells from 6- to 7-wk-old mice or testicular slices from 3- to 4-mo-old roosters were incubated with varying doses of luteinizing hormones (LHs)/gonadotropins (GTHs) in Medium 199 containing isobutyl-methyl-xanthine and N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid buffer (pH 7.4) at 34 degrees C (mice) or 37 degrees C (roosters) for 4 hr under continuous aeration of 95% O2-5% CO2 in a Dubnoff incubator shaken at 100 cycles/min. Androgen in the medium was measured by radioimmunoassay. The results revealed that dose-related androgen formations were obtained both in rooster and mouse systems in response to stimulations of all LHs/GTHs tested. The mouse system was more responsive to mammalian LHs and placental GTHs, less responsive to LHs from chickens, frogs, and turtles, and extremely unresponsive to piscine GTHs. In contrast, the rooster system was highly responsive to LHs from both mammals and chickens in androgen formation; it was also responsive to LHs from turtles and frogs as well as to piscine GTHs, although with relatively lower sensitivity. The rooster testis system is thus suitable for in vitro bioassay of LHs/GTHs from virtually all vertebrate classes, whereas the mouse testis system is more suitable for bioassay of mammalian LHs and placental GTHs. The differential androgen formation potencies of the diverse vertebrate GTHs in testis systems between roosters and mice indicate that a divergence exists in the testicular receptors between the two animal species.(ABSTRACT TRUNCATED AT 250 WORDS)     

Optogenetic Recruitment of Dorsal Raphe Serotonergic Neurons Acutely Decreases Mechanosensory Responsivity in Behaving Mice

The inhibition of sensory responsivity is considered a core serotonin function, yet this hypothesis lacks direct support due to methodological obstacles. We adapted an optogenetic approach to induce acute, robust and specific firing of dorsal raphe serotonergic neurons. In vitro, the responsiveness of individual dorsal raphe serotonergic neurons to trains of light pulses varied with frequency and intensity as well as between cells, and the photostimulation protocol was therefore adjusted to maximize their overall output rate. In vivo, the photoactivation of dorsal raphe serotonergic neurons gave rise to a prominent light-evoked field response that displayed some sensitivity to a 5-HT1A agonist, consistent with autoreceptor inhibition of raphe neurons. In behaving mice, the photostimulation of dorsal raphe serotonergic neurons produced a rapid and reversible decrease in the animals'' responses to plantar stimulation, providing a new level of evidence that serotonin gates sensory-driven responses.     

Strain differences in the amphibian chytrid Batrachochytrium dendrobatidis and non-permanent, sub-lethal effects of infection

The chytrid fungus Batrachochytrium dendrobatidis (Bd) is likely the cause of numerous recent amphibian population declines worldwide. While the fungus is generally highly pathogenic to amphibians, hosts express a wide range of responses to infection, probably due to variation among hosts and environmental conditions, but possibly also due to variation in Bd. We investigated variation in Bd by exposing standardized host groups to 2 Bd strains in a uniform environment. All exposed frogs became infected, but subsequent lethal and sub-lethal (weight loss) responses differed among groups. These results demonstrate variation in Bd and suggest variation occurs even at small geographical scales, likely explaining some of the variation in host responses. With lower than expected mortality among infected frogs, we continued our study opportunistically to determine whether or not frogs could recover from chytridiomycosis. Using heat, we cleared infection from half of the surviving frogs, leaving the other half infected, then continued to monitor mortality and weight. Mortality ceased among disinfected frogs but continued among infected frogs. Disinfected frogs gained weight significantly more than infected frogs, to the point of becoming indistinguishable from controls, demonstrating that at least some of the effects of sub-lethal chytridiomycosis on hosts can be non-permanent and reversible.     

The Lower Cretaceous Patuxent Formation Ichnofauna of Virginia

The vertebrate fauna from the Lower Cretaceous Patuxent Formation of Virginia is composed of a single partial fish impression from the James River at Dutch Gap and a diverse tetrapod ichnofauna from near Fredericksburg that includes trace fossils made by frogs, turtles, theropods, sauropods, ankylosaurs, and ornithopods. The footprints occur on overbank deposits preserved locally within a fluvial braided-stream sequence that formed near the western border of the Early Cretaceous Atlantic Coastal Plain.