Convergently evolved muscle architecture enables high-performance ballistic movement in salamanders

Elastically powered ballistic movements, such as tongue projection, are common in nature, likely due to benefits such as increased acceleration and distance of movement, and decreased thermal sensitivity imparted by elastic mechanisms. Within Plethodontidae, both muscle-powered and elastically powered ballistic tongue projection occur. Thus, we examine how elastically powered ballistic tongue projection morphology has evolved from muscle powered projection at the level of the projector muscles (m. subarcualis rectus [SAR]). We find that two main SAR morphologies have evolved within Plethodontidae. The first SAR morphology is conducive to elastically powered ballistic projection. This ballistic SAR morphology has evolved multiple, independent times within Plethodontidae, and results from the correlated evolution of several traits including increased collagen aponeuroses, larger SAR muscles, and the loss of inner myofibers attaching directly to the tongue skeleton. While the independent evolution of ballistic SAR morphology has arrived at a similar anatomical design, other tongue structures such as tongue attachment and skeleton folding type varies among species with a ballistic SAR morphology. The second morphology is conducive to muscle-powered projection and is similar to morphology found in an outgroup, Salamandridae. The SAR of these species have inner myofibers that attach to the tongue skeleton, limiting projection distance, coupled with reduced collagen aponeuroses present in relatively small projector muscles. This SAR morphology has likely been retained from ancestors or may be related to feeding ecology. Overall, a ballistic SAR morphology has evolved repeatedly and independently due to the correlated evolution of several SAR traits, including the loss of inner myofibers, which is likely a defining feature of ballistic projection.     

Rapid response to habitat restoration by the perennial Primula veris as revealed by demographic monitoring

The demography of Primula veris, a typical species of the species-rich Mesobromion grasslands, was investigated at two contrasting habitats in Eastern Belgium. Both a forested site and a clear-cut parcel were part of formerly larger calcareous grassland areas. By monitoring the demographic response of the target species, this study attempts to clarify the differences in fecundity, growth and survival between a restored and degraded calcareous grassland site. A study of plant traits showed a decrease in number of flowers, inflorescence stalks and plant size under a closing tree canopy. We surveyed individuals in permanent plots between 1999 and 2001. At the forested site, a first sign of decline included lower proportions of flowering individuals and afterwards an increase in mortality was found. Alternatively, removing canopy resulted in an immediate flowering response and both increased growth and seedling recruitment the year after. At both sites, survival rates strongly depended on the initial state, location and year. Projection matrix analysis revealed large differences in modelled population growth rates between sites and years. Under closing canopy the species showed only slow population decline, but the decrease was larger in the last survey year when higher mortality affected the number of reproductive individuals. Transitions between stages with the highest impact on population growth rate were identified by elasticity analysis. If calcareous grassland is forested, survival of reproductive adults is predicted to be very important for conservation of P. veris populations. However, seedling recruitment needs to be raised to guarantee long-term persistence of the populations. On the other hand, clear-felling of a site for restoration of species-rich calcareous grasslands may result in a rapid recovery of certain species.     

Fluorescence lifetime optical projection tomography

We describe a quantitative fluorescence projection tomography technique which measures the 3‐D fluorescence lifetime distribution in optically cleared specimens up 1 cm in diameter. This is achieved by acquiring a series of wide‐field time‐gated images at different relative time delays with respect to a train of excitation pulses, at a number of projection angles. For each time delay, the 3‐D time‐gated intensity distribution is reconstructed using a filtered back projection algorithm and the fluorescence lifetime subsequently determined for each reconstructed horizontal plane by iterative fitting to a mono‐exponential decay. Due to its inherently ratiometric nature, fluorescence lifetime is robust against intensity based artefacts as well as producing a quantitative measure of the fluorescence signal. We present a 3‐D fluorescence lifetime reconstruction of a mouse embryo labelled with an alexa‐488 conjugated antibody targeted to the neurofilament, which clearly differentiates between the extrinsic label and the autofluorescence, particularly from the heart and dorsal aorta. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Demographic factors affecting population growth in giant gartersnakes

Demographic models provide insight into which vital rates and life stages contribute most to population growth. Integral projection models (IPMs) offer flexibility in matching model structure to a species’ demography. For many rare species, data are lacking for key vital rates, and uncertainty might dissuade researchers from attempting to build a demographic model. We present work that highlights how the implications of uncertainties and unknowns can be explored by building and analyzing alternative models. We constructed IPMs for the threatened giant gartersnake (Thamnophis gigas) based on published studies to determine where management efforts could be targeted to have the greatest effect on population persistence and what unknowns remain for future research. Given uncertainty in the survival of snakes during their first year, and in the form of the size-survival relationship, we modeled a range of scenarios and evaluated where models agree about factors influencing population growth and where discrepancies exist. For most scenarios, the survival of large adult females had the greatest influence on population growth, but the relative importance of juvenile versus adult somatic growth for population growth was dependent on the recruitment probability and the shape of the size-survival function. More data on temporal variation and covariance among vital rates would improve stochastic models for the giant gartersnake. This paper demonstrates the effectiveness of IPMs for studying the demography of reptiles and the value of the model-building process for formalizing what is known and unknown about the demography of rare species. Published 2019. This article is a U.S. Government work and is in the public domain in the USA.     

Projecting the impacts of rising seawater temperatures on the distribution of seaweeds around Japan under multiple climate change scenarios

Seaweed beds play a key role in providing essential habitats and energy to coastal areas, with enhancements in productivity and biodiversity and benefits to human societies. However, the spatial extent of seaweed beds around Japan has decreased due to coastal reclamation, water quality changes, rising water temperatures, and heavy grazing by herbivores. Using monthly mean sea surface temperature (SST) data from 1960 to 2099 and SST‐based indices, we quantitatively evaluated the effects of warming seawater on the spatial extent of suitable versus unsuitable habitats for temperate seaweed Ecklonia cava, which is predominantly found in southern Japanese waters. SST data were generated using the most recent multiple climate projection models and emission scenarios (the Representative Concentration Pathways or RCPs) used in the Coupled Model Intercomparison Project phase 5 (CMIP5). In addition, grazing by Siganus fuscescens, an herbivorous fish, was evaluated under the four RCP simulations. Our results suggest that continued warming may drive a poleward shift in the distribution of E. cava, with large differences depending on the climate scenario. For the lowest emission scenario (RCP2.6), most existing E. cava populations would not be impacted by seawater warming directly but would be adversely affected by intensified year‐round grazing. For the highest emission scenario (RCP8.5), previously suitable habitats throughout coastal Japan would become untenable for E. cava by the 2090s, due to both high‐temperature stress and intensified grazing. Our projections highlight the importance of not only mitigating regional warming due to climate change, but also protecting E. cava from herbivores to conserve suitable habitats on the Japanese coast.     

Integrated Application of Multiomics Strategies Provides Insights Into the Environmental Hypoxia Response in Pelteobagrus vachelli Muscle

Increasing pressures on aquatic ecosystems because of pollutants, nutrient enrichment, and global warming have severely depleted oxygen concentrations. This sudden and significant lack of oxygen has resulted in persistent increases in fish mortality rates. Revealing the molecular mechanism of fish hypoxia adaptation will help researchers to find markers for hypoxia induced by environmental stress. Here, we used a multiomics approach to identify several hypoxia-associated miRNAs, mRNAs, proteins, and metabolites involved in diverse biological pathways in the muscles of Pelteobagrus vachelli. Our findings revealed significant hypoxia-associated changes in muscles over 4 h of hypoxia exposure and discrete tissue-specific patterns. We have previously reported that P. vachelli livers exhibit increased anaerobic glycolysis, heme synthesis, erythropoiesis, and inhibit apoptosis when exposed to hypoxia for 4 h. However, the opposite was observed in muscles. According to our comprehensive analysis, fishes show an acute response to hypoxia, including activation of catabolic pathways to generate more energy, reduction of biosynthesis to decrease energy consumption, and shifting from aerobic to anaerobic metabolic contributions. Also, we found that hypoxia induced muscle dysfunction by impairing mitochondrial function, activating inflammasomes, and apoptosis. The hypoxia-induced mitochondrial dysfunction enhanced oxidative stress, apoptosis, and further triggered interleukin-1β production via inflammasome activation. In turn, interleukin-1β further impaired mitochondrial function or apoptosis by suppressing downstream mitochondrial biosynthesis–related proteins, thus resulting in a vicious cycle of inflammasome activation and mitochondrial dysfunction. Our findings contribute meaningful insights into the molecular mechanisms of hypoxia, and the methods and study design can be utilized across different fish species.     

Population parameters and growth of Riptortus pedestris (Fabricius) (Hemiptera: Alydidae) under elevated CO2 concentrations

We evaluated the direct effects of three different CO₂ concentrations (400, 600 and 1,000 ppm) on the population parameters and growth of the bean bug, Riptortus pedestris, while being fed on soybean. The raw life history data from R. pedestris was analyzed using an age‐stage, two‐sex life table to take the viable development rate among individuals into account. Based on the age‐stage, two‐sex life table analysis, the population projections of R. pedestris provide the stage structure and variability of the population growth under different CO₂ treatments. Our results showed significantly shorter immature durations and higher pre‐adult survival rate under elevated CO₂ (1,000 ppm) than those under ambient CO₂ (400 ppm). The population of R. pedestris reared under elevated CO₂ conditions showed higher intrinsic and finite rates of increase but a lower mean generation time than R. pedestris reared under ambient CO₂ conditions. Our results show the population parameters and growth of R. pedestris are influenced by increased CO₂ relative to ambient CO₂ treatment. Further studies on the long‐term direct effects of different CO₂ levels on R. pedestris are essential to understand their population dynamics and to establish appropriate management strategies.     

Activity-driven sharpening of the retinotectal projection in goldfish: Development under stroboscopic illumination prevents sharpening

Blocking or synchronizing activity during regeneration of the retinotectal projection prevents both the sharpening of the retinotopic map recorded on tectum and the refinement of the structure of individual arbors within the plane of the map, and this refinement is triggered by N-methyl-d-aspartate (NMDA) receptors. We tested whether activity-driven refinement also occurs during development of the projection in larval and young adult goldfish. Shortly after hatching, larval goldfish were placed into tanks within light-tight chambers illuminated by a xenon strobe at 1 Hz for 14 h of each daily cycle. Fish were reared for 1.5–2 years, until large enough to record in our retinotectal mapping apparatus (6 cm length). Age- and size-matched controls had normal maps with multiunit receptive fields (MURFs) recorded at each tectal point of 10.8° (0.16 S.E.M., n = 5), whereas the strobe-reared fish had only roughly retino-topic maps with much enlarged MURFs averaging 26.7° (1.41 S.E.M., n = 5). This enlargement represents an abnormal convergence onto each tectal point, as the maps failed to sharpen during development. The arbors of individual retinal axons were stained with horseradish peroxidase (HRP) in larval fish and in adult strobereared and control fish. They were drawn with camera lucida from tectal whole mounts, and analyzed for spatial extent in the plane of the retinotopic map, order of branching, number of branch endings, depth of termination, and caliber of the parent axon. Arbors from larval fish (1–2 weeks) were small (approximately 50 × 40 μm) with less than 10 branches, occupied a single strata, and could not be separated into different classes by caliber of axon. The 87 arbors stained in control adult fish (6 cm long) were much like previously examined adult arbors, with those from fine, medium, and coarse axons averaging 115, 166, and 194 μm in extent, respectively, and having 17–24 branch endings. The 110 arbors from 12 strobe-reared fish were often abnormal. Although the fasciculation was normal, the extrafascicular routes were abnormal with reversing turns. The axons often had branches along their course, and these branches were scattered across a wider extent, rather than forming a distinct cluster. In contrast, neither the number of branches nor the depths of termination was significantly changed in any group. The coarse caliber arbors were most abnormal, being 64% longer and 30% wider than controls. The fine caliber arbors were also significantly larger by about 20%, but the medium caliber arbors were not enlarged. The enlarged arbors partially account for the unsharpened electrophysiological maps. Together the results show that during development, as well as during regeneration, the retinotectal map is subject to an activity-driven sharpening process. © 1993 John Wiley & Sons, Inc.     

Native insect herbivory overwhelms context dependence to limit complex invasion dynamics of exotic weeds

Understanding the role of consumers in density‐dependent plant population dynamics is a long‐standing goal in ecology. However, the generality of herbivory effects across heterogeneous landscapes is poorly understood due to the pervasive influence of context‐dependence. We tested effects of native insect herbivory on the population dynamics of an exotic thistle, Cirsium vulgare, in a field experiment replicated across eight sites in eastern Nebraska. Using hierarchical Bayesian analysis and density‐dependent population models, we found potential for explosive low‐density population growth (λ > 5) and complex density fluctuations under herbivore exclusion. However, herbivore access drove population decline (λ < 1), suppressing complex fluctuations. While plant–herbivore interaction outcomes are famously context‐dependent, we demonstrated that herbivores suppress potentially invasive populations throughout our study region, and this qualitative outcome is insensitive to environmental context. Our novel use of Bayesian demographic modelling shows that native insect herbivores consistently prevent hard‐to‐predict fluctuations of weeds in environments otherwise susceptible to invasion.     

Recruitment of APPL1 to ubiquitin-rich aggresomes in response to proteasomal impairment

Inhibitors of proteasomes have been shown to affect endocytosis of multiple membrane receptors, in particular at the step of cargo sorting for lysosomal degradation. Here we demonstrate that the inhibition of proteasomes causes specific redistribution of an endosomal adaptor APPL1, which undergoes initial solubilization from APPL endosomes followed by clustering in the perinuclear region. MG132 treatment decreases APPL1 labeling of endosomes while the staining of the canonical early endosomes with EEA1 remains unaffected. Upon prolonged treatment with proteasome inhibitors, endogenous APPL1 localizes to the site of aggresome formation, with perinuclear APPL1 clusters encapsulated within a vimentin cage and co-localizing with aggregates positive for ubiquitin. The clustering of APPL1 is concomitant with increased ubiquitination and decreased solubility of this protein. We determined that the ubiquitin ligase Nedd4 enhances polyubiquitination of APPL1, and the ubiquitin molecules attached to APPL1 are linked through lysine-63. Taken together, these results add APPL1 to only a handful of endogenous cellular proteins known to be recruited to aggresomes induced by proteasomal stress. Moreover, our studies suggest that the proteasome inhibitors that are already in clinical use affect the localization, ubiquitination and solubility of APPL1.