Rana Pipiens Research Articles

Toxicity of the conventional energetics TNT and RDX relative to new insensitive munitions constituents DNAN and NTO in Rana pipiens tadpoles.

An initiative within the US military is targeting the replacement of traditional munitions constituents with insensitive munitions to reduce risk of accidental detonation. The purpose of the present study was to comparatively assess toxicity of the traditional munitions constituents 2,4,6-trinitrotoluene (TNT) and 1,3,5-trinitroperhydro-1,3,5-triazine (RDX) with the new insensitive munitions constituents 2,4-dinitroanisole (DNAN) and 3-nitro-1,2,4-triazol-5-one (NTO). The following exposure durations were performed with Rana pipiens (leopard frog) tadpoles: TNT and DNAN, 96 h and 28 d; RDX, 10 d and 28 d; NTO, 28 d. The 96-h 50% lethal concentration (LC50) values and 95% confidence intervals for TNT and DNAN were 4.4 mg/L (4.2 mg/L, 4. 7 mg/L) and 24.3 mg/L (21.3 mg/L, 27.6 mg/L), respectively. No significant impacts on survival were observed in the 10-d exposure to RDX up to 25.3 mg/L. Effects on tadpole swimming distance were observed with a lowest-observed-effect concentration (LOEC) of 5.9 mg/L RDX. In the 28-d exposures, the LOECs for survival for TNT, DNAN, and NTO were 0.003 mg/L, 2.4 mg/L, and 5.0 mg/L, respectively. No significant mortality was observed in the RDX chronic 28-d exposure up to the highest treatment level tested of 28.0 mg/L. Neither tadpole developmental stage nor growth was significantly affected in any of the 28-d exposures. Rana pipiens were very sensitive to chronic TNT exposure, with an LOEC 3 orders of magnitude lower than those for insensitive munitions constituents DNAN and NTO.

Bioaccumulation kinetics of the conventional energetics TNT and RDX relative to insensitive munitions constituents DNAN and NTO in Rana pipiens tadpoles.

The manufacturing of explosives and their loading, assembling, and packing into munitions for use in testing on training sites or battlefields has resulted in contamination of terrestrial and aquatic sites that may pose risk to populations of sensitive species. The bioaccumulative potential of the conventional explosives 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and of the insensitive munitions (i.e., less shock sensitive) compound 2,4-dinitroanisole (DNAN) were assessed using the Northern leopard frog, Rana pipiens. Trinitrotoluene entering the organism was readily biotransformed to aminodinitrotoluenes, whereas no transformation products were measured for RDX or DNAN. Uptake clearance rates were relatively slow and similar among compounds (1.32-2.19 L kg(-1) h(-1) ). Upon transfer to uncontaminated water, elimination rate was very fast, resulting in the prediction of fast time to approach steady state (5 h or less) and short elimination half-lives (1.2 h or less). A preliminary bioconcentration factor of 0.25 L kg(-1) was determined for the insensitive munitions compound 3-nitro-1,2,4-trizole-5-one (NTO) indicating negligible bioaccumulative potential. Because of the rapid elimination rate for explosives, tadpoles inhabiting contaminated areas are expected to experience harmful effects only if under constant exposure conditions given that body burdens can rapidly depurate preventing tissue concentrations from persisting at levels that may cause detrimental biological effects.

Lesser of two evils? Foraging choices in response to threats of predation and parasitism.

Predators have documented post-encounter (density-mediated) effects on prey but their pre-encounter impacts, including behavioural alterations, can be substantial as well. While it is increasingly evident that this “ecology of fear” is important to understand for natural enemy-victim relationships, fear responses of hosts to the threat of infection by a parasite are relatively unknown. We examined larval amphibian (Lithobates pipiens) foraging choices by experimentally manipulating the presence of cues relating to predator (larval odonate) or parasite (the trematode Ribeiroia ondatrae) threats. Tadpoles avoided foraging where predator or parasite cues were present; however, they did not treat these as equal hazards. When both threats were simultaneously present, tadpoles strongly preferred to forage under the threat of parasitism compared to predation, likely driven by their relative lethality in our study. Our results indicate that altered spatial use is an important anti-parasite behaviour, and demonstrate that parasite avoidance can affect foraging in a manner similar to predators, warranting greater study of the pre-encounter effects of this enemy type.

The frog inner ear: picture perfect?

Many recent accounts of the frog peripheral auditory system have reproduced Wever's (1973) schematic cross-section of the ear of a leopard frog. We sought to investigate to what extent this diagram is an accurate and representative depiction of the anuran inner ear, using three-dimensional reconstructions made from serial sections of Rana pipiens, Eleutherodactylus limbatus and Xenopus laevis. In Rana, three discrete contact membranes were found to separate the posterior otic (=endolymphatic) labyrinth from the periotic (=perilymphatic) system: those of the amphibian and basilar recesses and the contact membrane of the saccule. The amphibian 'tegmentum vasculosum' was distinguishable as a thickened epithelial lining within a posterior recess of the superior saccular chamber. These features were also identified in Eleutherodactylus, but in this tiny frog the relative proportions of the semicircular canals and saccule resemble those of ranid tadpoles. There appeared to be a complete fluid pathway between the right and left periotic labyrinths in this species, crossing the cranial cavity. Xenopus lacks a tegmentum vasculosum and a contact membrane of the saccule; the Xenopus ear is further distinguished by a lateral passage separating stapes from periotic cistern and a more direct connection between periotic cistern and basilar recess. The basilar and lagenar recesses are conjoined in this species. Wever's diagram of the inner ear of Rana retains its value for diagrammatic purposes, but it is not anatomically accurate or representative of all frogs. Although Wever identified the contact membrane of the saccule, most recent studies of frog inner ear anatomy have overlooked both this and the amphibian tegmentum vasculosum. These structures deserve further attention.

Nematode parasites and leukocyte profiles of Northern Leopard Frogs,Rana pipiens: location, location, location

Globally, amphibians face a variety of anthropogenic stresses that include exposure to contaminants such as agricultural pesticides. Pesticides may negatively affect amphibian immune systems, concomitantly increasing susceptibility to parasitism. We quantified nematodes and evaluated leukocyte profiles of Northern Leopard Frogs (Rana pipiens Schreber, 1782) collected from five wetlands in southwestern Quebec, Canada, that spanned a gradient of pesticide exposure. Three taxa of nematode parasites (Rhabdias ranae Walton, 1929, genus Oswaldocruzia Travassos, 1917, and genus Strongyloides Grassi, 1879) were sufficiently numerous for detailed evaluation. When all frogs were pooled, frog size was negatively correlated with nematode species richness, abundances of each of the three nematode species, and densities of three different leukocytes. When all frogs were pooled, there was strong evidence of both negative and positive associations between pairs of parasite species. However, none of the previous relationships was significant within wetlands. Our results reveal strong spatial organization of amphibian–parasite communities and illustrate the importance of controlling for sampling locale in evaluating host–parasite associations. Finally, although several response variables varied significantly among wetlands, causes of this variation did not appear to be related to variation in nematode parasitism or pesticide exposure.

Toe Clipping Does Not Affect the Survival of Leopard Frogs (Rana pipiens)

Toe clipping is commonly used to identify individual frogs in mark–recapture studies. However, Institutional Animal Care and Use Committees (IACUCs) often have reservations about approving studies that include toe clipping as a way to mark animals. Previous studies indicate that recapture rates of terrestrial frogs and toads decrease with increasing numbers of toes clipped. It is not known whether these decreases are due to decreased survival rates, increased avoidance of recapture, or some other reason. The goal of this paper is to provide IACUCs with a quantitative analysis of the effects of toe clipping on survival so that they can make informed, data-based decisions when approving studies using this method, rather than basing decisions on anthropomorphic biases. We conducted a large (n = 100) laboratory experiment to determine the effects of toe clipping on the frog Rana pipiens. We found no evidence that the number of toes clipped affected either the survival rate of Leopard Frogs or the number of ...

Onconase induces autophagy sensitizing pancreatic cancer cells to gemcitabine and activates Akt/mTOR pathway in a ROS-dependent manner

Onconase® (ONC) is a member of the RNase super-family that is secreted in oocytes and early embryos of Rana pipiens. Over the last years, research interest about this small and basic frog RNase, also called ranpirnase, constantly increased because of its high cytotoxicity and anticancer properties. Onconase is currently used in clinical trials for cancer therapy; however, the precise mechanisms determining cytotoxicity in cancer cells have not yet been fully investigated. In the present manuscript, we evaluate the antitumoral property of onconase in pancreatic adenocarcinoma cells and in non-tumorigenic cells as a control. We demonstrate that ONC stimulates a strong antiproliferative and proapoptotic effect in cancer cells by reporting for the first time that ONC triggers Beclin1-mediated autophagic cancer cell death. In addition, ONC inhibits the expression of mitochondrial uncoupling protein 2 (UCP2) and of manganese-dependent superoxide dismutase (MnSOD) triggering mitochondrial superoxide ion production. ONC-induced reactive oxygen species (ROS) are responsible for Akt/mTOR pathway stimulation determining the sensitivity of cancer cells to mTOR inhibitors and lessening autophagic stimulation. This indicates ROS/Akt/mTOR axis as a strategy adopted by cancer cells to reduce ONC-mediated cytotoxic autophagy stimulation. In addition, we demonstrate that ONC can sensitize pancreatic cancer cells to the standard chemotherapeutic agent gemcitabine allowing a reduction of drug concentration when used in combination settings, thus suggesting a lowering of chemotherapy-related side effects. Altogether, our results shed more light on the mechanisms lying at the basis of ONC antiproliferative effect in cancer cells and support its potential use to develop new anticancer strategies.

The mechanics of elastic loading and recoil in anuran jumping.

Many animals use catapult mechanisms to produce extremely rapid movements for escape or prey capture, resulting in power outputs far beyond the limits of muscle. In these catapults, muscle contraction loads elastic structures, which then recoil to release the stored energy extremely rapidly. Many arthropods employ anatomical 'catch mechanisms' to lock the joint in place during the loading period, which can then be released to allow joint motion via elastic recoil. Jumping vertebrates lack a clear anatomical catch, yet face the same requirement to load the elastic structure prior to movement. There are several potential mechanisms to allow loading of vertebrate elastic structures, including the gravitational load of the body, a variable mechanical advantage, and moments generated by the musculature of proximal joints. To test these hypothesized mechanisms, we collected simultaneous 3D kinematics via X-ray Reconstruction of Moving Morphology (XROMM) and single-foot forces during the jumps of three Rana pipiens. We calculated joint mechanical advantage, moment and power using inverse dynamics at the ankle, knee, hip and ilio-sacral joints. We found that the increasing proximal joint moments early in the jump allowed for high ankle muscle forces and elastic pre-loading, and the subsequent reduction in these moments allowed the ankle to extend using elastic recoil. Mechanical advantage also changed throughout the jump, with the muscle contracting against a poor mechanical advantage early in the jump during loading and a higher mechanical advantage late in the jump during recoil. These 'dynamic catch mechanisms' serve to resist joint motion during elastic loading, then allow it during elastic recoil, functioning as a catch mechanism based on the balance and orientation of forces throughout the limb rather than an anatomical catch.

Soft catch prepares power for frog leaps

When it comes to a list of the top animal jumpers, frogs are right up there, along with other members of the superleague, such as grasshoppers and fleas. Yet, how frogs pull off their remarkable ballistic feat was unclear. Henry Astley from Brown University, USA, explains that insects wind up for a jump by locking their legs in place while their muscles pull on an elastic piece of exoskeleton that stores energy ready for release when the catch is unleashed to power the leg's thrust. However, Astley and Tom Roberts were puzzled how frogs could produce the same explosive feat in a soft body. ‘Jumping vertebrates lack a clear anatomical catch, yet face the same requirement to load the elastic structure prior to movement,’ says Astley. Intrigued, the duo began filming the leaping amphibians with X-rays to find out more about how frogs power jumps (p. 4372).Inserting minute metal markers that show up well in X-ray movies into the hind legs of three frogs (Rana pipiens) and filming the animals as they let fly, Astley and Roberts also recorded the forces exerted on the ground by the frogs' feet. Then the duo reconstructed the way that the frogs unfurled their legs in the last 150 ms before push off and calculating the amount of power produced by the amphibian's ankle extensor (plantaris) muscle. Amazingly, this muscle, which powers leaps, was capable of producing over four times as much power (1352 W kg–1) as a normally contracting muscle (322 W kg–1), confirming that the animals were storing elastic energy and using it to power take-off like some other frogs.Next, Astley and Roberts analysed the leg movement reconstructions, and realised that the frogs were storing elastic energy at the ankle by changing their posture to alter the leverage and forces acting around the joint. Describing this ‘dynamic catch’ mechanism, Astley explains that elastic energy can be stored in the plantaris muscle during the preparation phase of the jump when the leverage acting at the ankle is poor and the forces acting on the bent legs – such as the ground reaction force – resist movements at the ankle. However, in the later stages of preparation, the forces that had resisted the ankle's movement fall to a point where the ground reaction force and leverage becomes great enough to release the energy stored in the plantaris muscle, launching the frog into the air.Explaining that the leg forces and poor leverage that resist ankle movements in the early stage of a jump are analogous to the mechanical catch that allows jumping insects to store elastic energy, the duo suspects that other animals may also be able to take advantage of this dynamic catch mechanism to produce impressive leaps. However, they point out that there are situations where this mechanism will not work – such as leg kicking, when there are no ground reaction forces to hold the limb steady during preparation.

Maternal body condition influences magnitude of anti-predator response in offspring.

Organisms exhibit plasticity in response to their environment, but there is large variation even within populations in the expression and magnitude of response. Maternal influence alters offspring survival through size advantages in growth and development. However, the relationship between maternal influence and variation in plasticity in response to predation risk is unknown. We hypothesized that variation in the magnitude of plastic responses between families is at least partly due to maternal provisioning and examined the relationship between maternal condition, egg provisioning and magnitude of plastic response to perceived predation risk (by dragonfly larvae: Aeshna spp.) in northern leopard frogs (Lithobates pipiens). Females in better body condition tended to lay more (clutch size) larger (egg diameter) eggs. Tadpoles responded to predation risk by increasing relative tail depth (morphology) and decreasing activity (behaviour). We found a positive relationship between morphological effect size and maternal condition, but no relationship between behavioural effect size and maternal condition. These novel findings suggest that limitations imposed by maternal condition can constrain phenotypic variation, ultimately influencing the capacity of populations to respond to environmental change.

Do hormone-modulating chemicals impact on reproduction and development of wild amphibians?

Globally, amphibians are undergoing a precipitous decline. At the last estimate in 2004, 32% of the approximately 6000 species were threatened with extinction and 43% were experiencing significant declines. These declines have been linked with a wide range of environmental pressures from habitat loss to climate change, disease and pollution. This review evaluates the evidence that endocrine-disrupting contaminants (EDCs) - pollutants that affect hormone systems - are impacting on wild amphibians and contributing to population declines. The review is limited to anurans (frogs and toads) as data for effects of EDCs on wild urodeles (salamanders, newts) or caecilians (limbless amphibians) are extremely limited. Evidence from laboratory studies has shown that a wide range of chemicals have the ability to alter hormone systems and affect reproductive development and function in anurans, but for the most part only at concentrations exceeding those normally found in natural environments. Exceptions can be found for exposures to the herbicide atrazine and polychlorinated biphenyls in leopard frogs (Rana pipiens) and perchlorate in African clawed frogs (Xenopus laevis). These contaminants induce feminising effects on the male gonads (including 'intersex' - oocytes within testes) at concentrations measured in some aquatic environments. The most extensive data for effects of an EDC in wild amphibian populations are for feminising effects of atrazine on male gonad development in regions across the USA. Even where strong evidence has been provided for feminising effects of EDCs, however, the possible impact of these effects on fertility and breeding outcome has not been established, making inference for effects on populations difficult. Laboratory studies have shown that various chemicals, including perchlorate, polychlorinated biphenyls and bromodiphenylethers, also act as endocrine disrupters through interfering with thyroid-dependent processes that are fundamental for amphibian metamorphosis. Perchlorate has also been shown to induce these effects in wild anuran populations from perchlorate-contaminated environments. Overall, the published data available suggest that some health effects observed in wild anuran populations, most notably intersex, likely have a chemical aetiology; however they derive only from very few anuran species and for a few pesticides at field sites in the USA. To understand better the impacts of EDCs on wild anuran populations, as well as other amphibian groups, assessment of fertility in exposed animals are required. Development of non-destructive biomarkers that are indicative of specific EDC-effect mechanisms are also needed to allow the study of vulnerable populations. This will help to distinguish the effects of EDCs from other environmental and/or genetic influences on development and reproduction.

New disease records for hatchery-reared sturgeon. I. Expansion of frog virus 3 host range into Scaphirhynchus albus.

In 2009, juvenile pallid sturgeon Scaphirhynchus albus, reared at the Blind Pony State Fish Hatchery (Missouri, USA) to replenish dwindling wild stocks, experienced mass mortality. Histological examination revealed extensive necrosis of the haematopoietic tissues, and a virus was isolated from affected organs in cell culture and then observed by electron microscopy. Experimental infection studies revealed that the virus is highly pathogenic to juvenile pallid sturgeon, one of several species of sturgeon currently listed as Endangered. The DNA sequence of the full length major capsid protein gene of the virus was identical to that of the species Frog virus 3 (FV3), the type species for the genus Ranavirus, originally isolated from northern leopard frog Lithobates pipiens. Although FV3 infections and epizootics in amphibians and reptiles are well documented, there is only 1 prior report of a natural infection of FV3 in fish. Our results illustrate the broad potential host range for FV3, with the known potential to cause significant mortality in poikilothermic vertebrates across 3 taxonomic classes including bony fishes, anuran and caudate amphibians, and squamate and testudine reptiles.

A fusogenic dengue virus-derived peptide enhances antitumor efficacy of an antibody-ribonuclease fusion protein targeting the EGF receptor.

Due to its frequent overexpression in a variety of solid tumors the epidermal growth factor receptor (EGFR) is a well-established target for therapeutic interventions in epithelial cancers. In order to target EGFR in head and neck cancer, we have generated a ribonuclease (RNase) fusion protein comprising a humanized anti-EGFR antibody single-chain Fv fragment (scFv) and Ranpirnase, an RNase from Rana pipiens. Fusion of Ranpirnase to the N-terminus of the scFv via a flexible glycine-serine linker (G4S)3 resulted in very poor cytotoxicity of the fusion protein. As endosomal accumulation and lysosomal degradation have been reported to diminish the antitumor efficacy of ribonuclease or toxin-based immunoagents, we explored a fusion peptide from dengue virus that has been reported to be involved in the endosomal escape of the virus. This peptide was introduced as a linker between Ranpirnase and the scFv moiety. The modified immunoRNase exhibited exceptionally high cytotoxicity toward EGFR-expressing head and neck cell lines without affecting specificity. These results indicate that endosomal entrapment needs to be considered for Ranpirnase-based immunoagents and might be overcome by the use of tailored transduction domains from viral proteins.

Role of sediments in modifying the toxicity of two Roundup formulations to six species of larval anurans.

The role of sediment in modifying the toxicity of the original formulation of Roundup® and Roundup WeatherMAX® was examined in aqueous laboratory tests. Six species of anurans (Bufo fowleri, Hyla chrysoscelis, Rana catesbeiana, Rana clamitans, Rana sphenocephala, and Rana pipiens) were exposed at Gosner stage 25 to concentrations of the 2 herbicide formulations in 96-h, static, nonrenewal experiments in the presence and absence of sediment. All species tested had lower median lethal concentration values in water-only exposures of both formulations compared with exposures with sediment. Sediment significantly altered the potency slopes in all tests with the exceptions of H. chrysoscelis and R. clamitans when exposed to the original formulation of Roundup and H. chrysoscelis and R. sphenocephala when exposed to Roundup WeatherMAX. Thresholds were significantly different in all tests, including those in which potency slopes did not differ. Based on water-sediment exposures of the original formulation of Roundup, all 6 species tested had a margin of safety when compared with the predicted environmental concentration of the highest label application rate. Of the 6 species, 5 had a margin of safety when exposed to Roundup WeatherMAX. During incidental exposures in the field, sediments and organic matter present in aquatic systems provide significant sources of environmental ligands. If used according to label instructions, both herbicides should pose minimal risk to anuran amphibians in actual field applications. Environ Toxicol Chem 2014;33:2616-2620. © 2014 SETAC.

Capsaicin and N-arachidonoyl-dopamine (NADA) decrease tension by activating both cannabinoid and vanilloid receptors in fast skeletal muscle fibers of the frog.

Previous studies have indicated that vanilloid receptor (VR1) mRNA is expressed in muscle fibers. In this study, we evaluated the functional effects of VR1 activation. We measured caffeine-induced contractions in bundles of the extensor digitorum longus muscle of Rana pipiens. Isometric tension measurements showed that two VR1 agonists, capsaicin (CAP) and N-arachidonoyl-dopamine (NADA), reduced muscle peak tension to 57 ± 4 % and 71 ± 3% of control, respectively. The effect of CAP was partially blocked by a VR1 blocker, capsazepine (CPZ), but the effect of NADA was not changed by CPZ. Because NADA is able to act on cannabinoid receptors, which are also present in muscle fibers, we tested the cannabinoid antagonist AM281. We found that AM281 antagonized both CAP and NADA effects. AM281 alone reduced peak tension to 80 ± 6 % of control. With both antagonists, the CAP effect was completely blocked, and the NADA effect was partially blocked. These results provide pharmacological evidence of the functional presence of the VR1 receptor in fast skeletal muscle fibers of the frog and suggest that capsaicin and NADA reduce tension by activating both cannabinoid and vanilloid receptors.

Presence and colocalization of type-1 cannabinoid receptors with acetylcholine receptors in the motor end-plate of twitch skeletal muscle fibers in the frog.

Using polyclonal and monoclonal antibodies to visualize under a confocal microscope type-1 cannabinoid receptors (CB1) and acetylcholine (ACh) receptors, respectively, or α-bungarotoxin conjugated to Alexa-Fluor 555 for Ach receptors, we found that they colocalize on twitch muscle fibers in the frog (Rana pipiens). We show that both the CB1 and ACh receptors are present on the fast skeletal muscle motor end-plate. The CB1 receptor is present along the entire membrane of the muscle fiber, whereas the ACh receptor is expressed primarily at the motor end-plate. Analysis of the colocalization produced a cross-correlation coefficient of 0.519 ± 0.021 (n = 9) for both receptors at the muscle motor end-plate. This study suggests a close proximity between these two types of receptor proteins and that they could interact. CB1 could function at some stage of excitation-contraction coupling in these muscle fibers. However, further investigation is needed in order to clarify these issues.

Population of Northern Leopard Frogs (Rana Pipiens) Migrating Between the Ney Frog Pond and the Minnesota River Valley for Spring Breeding

Population of Northern Leopard Frogs (Rana Pipiens) Migrating Between the Ney Frog Pond and the Minnesota River Valley for Spring Breeding

Environmental dependency of amphibian-ranavirus genotypic interactions: evolutionary perspectives on infectious diseases.

The context-dependent investigations of host–pathogen genotypic interactions, where environmental factors are explicitly incorporated, allow the assessment of both coevolutionary history and contemporary ecological influences. Such a functional explanatory framework is particularly valuable for describing mortality trends and identifying drivers of disease risk more accurately. Using two common North American frog species (Lithobates pipiens and Lithobates sylvaticus) and three strains of frog virus 3 (FV3) at different temperatures, we conducted a laboratory experiment to investigate the influence of host species/genotype, ranavirus strains, temperature, and their interactions, in determining mortality and infection patterns. Our results revealed variability in host susceptibility and strain infectivity along with significant host–strain interactions, indicating that the outcome of an infection is dependent on the specific combination of host and virus genotypes. Moreover, we observed a strong influence of temperature on infection and mortality probabilities, revealing the potential for genotype–genotype–environment interactions to be responsible for unexpected mortality in this system. Our study thus suggests that amphibian hosts and ranavirus strains genetic characteristics should be considered in order to understand infection outcomes and that the investigation of coevolutionary mechanisms within a context-dependent framework provides a tool for the comprehensive understanding of disease dynamics.

Specialized postsynaptic morphology enhances neurotransmitter dilution and high-frequency signaling at an auditory synapse.

Sensory processing in the auditory system requires that synapses, neurons, and circuits encode information with particularly high temporal and spectral precision. In the amphibian papillia, sound frequencies up to 1 kHz are encoded along a tonotopic array of hair cells and transmitted to afferent fibers via fast, repetitive synaptic transmission, thereby promoting phase locking between the presynaptic and postsynaptic cells. Here, we have combined serial section electron microscopy, paired electrophysiological recordings, and Monte Carlo diffusion simulations to examine novel mechanisms that facilitate fast synaptic transmission in the inner ear of frogs (Rana catesbeiana and Rana pipiens). Three-dimensional anatomical reconstructions reveal specialized spine-like contacts between individual afferent fibers and hair cells that are surrounded by large, open regions of extracellular space. Morphologically realistic diffusion simulations suggest that these local enlargements in extracellular space speed transmitter clearance and reduce spillover between neighboring synapses, thereby minimizing postsynaptic receptor desensitization and improving sensitivity during prolonged signal transmission. Additionally, evoked EPSCs in afferent fibers are unaffected by glutamate transporter blockade, suggesting that transmitter diffusion and dilution, and not uptake, play a primary role in speeding neurotransmission and ensuring fidelity at these synapses.

Overwintering observations of Northern Leopard Frog (Lithobates pipiens) activity under ice

Overwintering observations of Northern Leopard Frog (Lithobates pipiens) activity under ice