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Whole-brain calcium imaging during physiological vestibular stimulation in larval zebrafish

During in vivo functional imaging, animals are head-fixed and thus deprived from vestibular inputs, which severely hampers the design of naturalistic virtual environments. To overcome this limitation, we developed a miniaturized ultra-stable light-sheet microscope that can be dynamically rotated during imaging along with a head-restrained zebrafish larva. We demonstrate that this system enables whole-brain functional imaging at single-cell resolution under controlled vestibular stimulation. We recorded for the first time the dynamic whole-brain response of a vertebrate to physiological vestibular stimulation. This development largely expands the potential of virtual-reality systems to explore complex multisensory-motor integration in 3D.

neuroscience

Physiological starvation increases EGF-Ras-MAPK pathway activity during C. elegans vulval induction

Studying how molecular pathways respond to ecologically relevant environmental variation is fundamental to understand organismal development and its evolution. Here we characterize how starvation modulates Caenorhabditis elegans vulval cell fate patterning - an environmentally sensitive process, with a nevertheless robust output. Past research has shown many vulval mutants affecting EGF-Ras-MAPK, Delta-Notch and Wnt pathways to be suppressed by environmental factors, such as starvation. Here we aimed to resolve previous, seemingly contradictory, observations on how starvation modulates levels of vulval induction. Using the strong starvation suppression of the Vulvaless phenotype of lin-3/egf reduction-of-function mutations as an experimental paradigm, we first tested for a possible involvement of the sensory system in relaying starvation signals to affect vulval induction: mutation of various sensory inputs, DAF-2/Insulin or DAF-7/TGF-{beta} signaling did not abolish lin-3(rf) starvation suppression. In contrast, nutrient deprivation induced by mutation of the intestinal peptide transporter gene pept-1 or the TOR pathway component rsks-1 (the orthologue of mammalian P70S6K) very strongly suppressed lin-3(rf) mutant phenotypes. Therefore, physiologically starved animals induced by these mutations tightly recapitulated the effects of external starvation on vulval induction. While both starvation and pept-1 RNAi were sufficient to increase Ras and Notch pathway activities in vulval cells, the highly penetrant Vulvaless phenotype of a tissue-specific null allele of lin-3 was not suppressed by either condition. This and additional results indicate that partial lin-3 expression is required for starvation to affect vulval induction. These results suggest a cross-talk between nutrient deprivation, TOR-S6K and EGF-Ras-MAPK signaling during C. elegans vulval induction.

genetics

A large-scale, standardized physiological survey reveals higher order coding throughout the mouse visual cortex

To understand how the brain processes sensory information to guide behavior, we must know how stimulus representations are transformed throughout the visual cortex. Here we report an open, large-scale physiological survey of neural activity in the awake mouse visual cortex: the Allen Brain Observatory Visual Coding dataset. This publicly available dataset includes cortical activity from nearly 60,000 neurons collected from 6 visual areas, 4 layers, and 12 transgenic mouse lines from 221 adult mice, in response to a systematic set of visual stimuli. Using this dataset, we reveal functional differences across these dimensions and show that visual cortical responses are sparse but correlated. Surprisingly, responses to different stimuli are largely independent, e.g. whether a neuron responds to natural scenes provides no information about whether it responds to natural movies or to gratings. We show that these phenomena cannot be explained by standard local filter-based models, but are consistent with multi-layer hierarchical computation, as found in deeper layers of standard convolutional neural networks.

neuroscience

Physiological Dynamic Compression Regulates Central Energy Metabolism in Primary Human Chondrocytes

Chondrocytes use the pathways of central metabolism to synthesize molecular building blocks and energy for cartilage homeostasis. An interesting feature of the in vivo chondrocyte environment is the cyclical loading generated in various activities (e.g. walking). However, it is unknown if central metabolism is altered by mechanical loading. We hypothesized that physiological dynamic compression alters central metabolism in chondrocytes to promote production of amino acid precursors for matrix synthesis. We measured the expression of central metabolites (e.g. glucose, its derivatives, and relevant co-factors) for primary human osteoarthritic chondrocytes in response to 0-30 minutes of compression. To analyze the data, we used principal components analysis and ANOVA simultaneous components analysis, as well as metabolic [fl]ux analysis. Compression induced metabolic responses consistent with our hypothesis. Additionally, these data show that chondrocyte samples from different patient donors exhibit different sensitivity to compression. Most important, we [fi]nd that grade IV osteoarthritic chondrocytes are capable of synthesizing non-essential amino acids and precursors in response to mechanical loading. These results suggest that further advances in metabolic engineering of chondrocyte mechanotransduction may yield novel translational strategies for cartilage repair.

systems biology

Gut bacterial diversity and physiological traits of Anastrepha fraterculus Brazilian-1 morphotype males are affected by antibiotic treatment

BackgroundThe interaction between gut bacterial symbionts and Tephritidae became the focus of several studies that showed that bacteria contributed to the nutritional status and the reproductive potential of its fruit fly hosts. Anastrepha fraterculus is an economically important fruit pest in South America. This pest is currently controlled by insecticides, which prompt the development of environmentally friendly methods such as the sterile insect technique (SIT). For SIT to be effective, a deep understanding of the biology and sexual behavior of the target species is needed. Although many studies have contributed in this direction, little is known about the composition and role of A. fraterculus symbiotic bacteria. In this study we tested the hypothesis that gut bacteria contribute to nutritional status and reproductive success of A. fraterculus males.\n\nMethodsWild and laboratory-reared males were treated with antibiotics (AB) and provided sugar (S) or sugar plus protein (S+P) as food sources. The effect of AB on the gut bacteria diversity was assessed through DGGE and sequencing of the V6-V9 variable region of the bacterial 16S rRNA gene.\n\nResultsAB affected the bacterial community of the digestive tract of A. fraterculus, in particular bacteria belonging to the Enterobacteriaceae family, which was the dominant bacterial group in the control flies (i.e., non-treated with AB). AB negatively affected parameters directly related to the mating success of laboratory males and their nutritional status. AB also affected males survival under starvation conditions. The effect of AB on the behaviour and nutritional status of the males depended on two additional factors: the origin of the males and the presence of a proteinaceous source in the diet.\n\nConclusionsOur results suggest that A. fraterculus males gut contain symbiotic organisms that are able to exert a positive contribution on A. fraterculus males fitness, although the physiological mechanisms still need further studies.

ecology

Characterization for Drought Tolerance and Physiological Efficiency in Novel Cytoplasmic Male Sterile Sources of Sunflower (Helianthus annuus L.)

AbstractSunflower is sensitive to drought and its hybrids have a limited cytoplasmic diversity. The wild cytoplasmic sources of sunflower are not well exploited to their potential for drought tolerance and hybrid development. In this respect, we carried out a Line x Tester based genetic study using 19 sunflower genotypes representing, 13 cytoplasmic male sterile (CMS) lines from wild and conventional sources, 2 maintainer lines, and 4 restorer lines. The CMS and maintainer lines were crossed with restorer lines to develop sixty one-way F1 hybrids. The parents and their hybrids were evaluated under two water regimes viz., normal irrigated and water stress. A total of twelve important plant descriptors were studied over a period of two years. The significant differences were observed between parents and hybrids in both water regimes. Hybrids were higher in average values for all the descriptors than parents. The role of female parent was more prominent in the expression of traits in hybrids as compared to male parents. The CMS sources varied significantly regarding seed yield per plant and other physiological traits. Proline content was three times higher in parents and their hybrids under water stress, and it was not correlated with any other descriptor. Accession CMS-PKU-2A was identified as the best general combiner for leaf area and specific leaf weight. Whereas, CMS-234A was the best general combiner for biological yield and photosynthetic efficiency under both the conditions. The cross combinations CMS-ARG-2A x RCR-8297, CMS-234A x P124R, and CMS-38A x P124R were found significant for biological yield, seed yield and oil content under both environments. Overall, this study provides useful information about the cytoplasmic effects on important sunflower traits and drought stress tolerance when used in the different combinations.

genetics

Anatomical and physiological foundations of cerebello-hippocampal interactions

Multiple lines of evidence suggest that functionally intact cerebello-hippocampal interactions are required for appropriate spatial processing. However, how the cerebellum anatomically and physiologically engages with the hippocampus to sustain such interactions remains unknown. Using rabies virus as retrograde transneuronal tracer, we reveal that the dorsal hippocampus receives input from topographically restricted and disparate regions of the cerebellum. By simultaneously recording local field potential from both the dorsal hippocampus and anatomically connected cerebellar regions, we additionally demonstrate that the two structures interact, in a behaviorally dynamic manner, through subregion-specific synchronization of neuronal oscillations in the 6-12Hz frequency range. Together, these results reveal a novel neural network macro-architecture through which we can understand how a brain region classically associated with motor control, the cerebellum, may influence hippocampal neuronal activity and related functions, such as spatial navigation.

neuroscience

Non-parametric physiological classification of retinal ganglion cells

Retinal ganglion cells, the sole output neurons of the retina, exhibit surprising diversity. A recent study reported over 30 distinct types in the mouse retina, indicating that the processing of visual information is highly parallelised in the brain. The advent of high density multi-electrode arrays now enables recording from many hundreds to thousands of neurons from a single retina. Here we describe a method for the automatic classification of large-scale retinal recordings using a simple stimulus paradigm and a spike train distance measure as a clustering metric. We evaluate our approach using synthetic spike trains, and demonstrate that major known cell types are identified in high-density recording sessions from the mouse retina with around 1000 retinal ganglion cells. A comparison across different retinas reveals substantial variability between preparations, suggesting pooling data across retinas should be approached with caution. As a parameter-free method, our approach is broadly applicable for cellular physiological classification in all sensory modalities.

neuroscience

The effect of physiological and measurement noise on the estimate of individual muscle force from indirect measurements of muscle activity

Several forward dynamics estimators have been proposed to quantify individual muscle force using non-invasive measurements of muscle activity. None of them, however, addresses the inaccuracy that arises when measurements are available only from a subset of the muscles involved in the movement under analysis. We present a novel estimator that integrates a forward dynamics estimation approach with knowledge of the optimal contraction strategy to obtain accurate estimates of individual muscle force when measurements of muscle activity are not available for all muscles. A following in-silico characterization showed that when trying to estimate forces form the forearm muscles acting around the wrist joint, our novel estimator is able to decrease the mean estimation bias by about 25% of the true value of muscle force. With a sensitivity analysis, we show that the model-based estimator is robust against physiological variability in muscle co-contraction strategy.

bioengineering

Effects of two centuries of global environmental variation on phenology and physiology of Arabidopsis thaliana

Intraspecific trait variation is caused by genetic and plastic responses to environment. This intraspecific diversity is captured in immense natural history collections, giving us a window into trait variation across continents and through centuries of environmental shifts. Here we tested if hypotheses based on life history and the leaf economics spectrum explain intraspecific trait changes across global spatiotemporal environmental gradients. We measured phenotypes on a 216-year time series of Arabidopsis thaliana accessions from across the native range and applied spatially varying coefficient models to quantify region-specific trends in trait coordination and trait responses to climate gradients. All traits exhibited significant change across space and/or through time. For example, {delta}15N decreased over time across much of the range and leaf C:N increased, consistent with predictions based on anthropogenic changes in land use and atmosphere. Plants were collected later in the growing season in more recent years in many regions, possibly because populations shifted toward more spring germination and summer flowering as opposed to fall germination and spring flowering. When climate variables were considered, collection dates were earlier in warmer years, while summer rainfall had opposing associations with collection date depending on regions. There was only a modest correlation among traits, indicating a lack of a single life history/physiology axis. Nevertheless, leaf C:N was low for summer- versus spring-collected plants, consistent with a life history-physiology axis from slow-growing winter annuals to fast-growing spring/summer annuals. Regional heterogeneity in phenotype trends indicates complex responses to spatiotemporal environmental gradients potentially due to geographic genetic variation and climate interactions with other aspects of environment. Our study demonstrates how natural history collections can be used to broadly characterize trait responses to environment, revealing heterogeneity in response to anthropogenic change.

ecology

The physiological butyrylcholinesterase tetramer is a dimer of dimers stabilized by a superhelical assembly

The quaternary structures of the cholinesterases, acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), are essential for their localisation and function. Of practical importance, BChE is a promising therapeutic candidate for intoxication by organophosphate nerve agents and insecticides, and for detoxification of addictive substances. Efficacy of the recombinant enzyme hinges on its having a long circulatory half-life; this, in turn, depends strongly on its ability to tetramerize. Here, we used cryo-electron microscopy (cryo-EM) to determine the structure of the highly glycosylated native BChE tetramer purified from human plasma at 5.7 [A]. Our structure reveals that the BChE tetramer is organised as a staggered dimer of dimers. Tetramerization is mediated by assembly of the C-terminal tryptophan amphiphilic tetramerization (WAT) helices from each subunit as a superhelical assembly around a central anti-parallel polyproline II helix (PRAD). The catalytic domains within a dimer are asymmetrically linked to the WAT/PRAD. In the resulting arrangement, the tetramerization domain is largely shielded by the catalytic domains, which may contribute to the stability of the HuBChE tetramer. Our cryo-EM structure reveals the basis for assembly of the physiological tetramers, and has implications for the therapeutic applications of HuBChE. This mode of tetramerization is seen only in the cholinesterases, and may provide a promising template for designing other proteins with improved circulatory residence times.

biophysics

Root endophytes improve physiological performance and yield in crops under salt stress by up-regulating the foliar sodium concentration

Increase in the soil salinity will be a conspicuous constraint for both native plant communities as well as several crops worldwide. In this context plant root-associated fungi appear as a new strategy to improve ecophysiological performance and yield of crops under abiotic stress. Here, we evaluated how the inoculation of fungal endophytes isolated from Antarctic plants can improve the ecophysiological performance in cultivars of tomato and lettuce, grown under different salt conditions. In addition, we assessed if the expression of the NHX1 gene that is related with salt tolerance is enhanced in presence of fungal endophytes. Finally, we evaluated the regulation the nutritional status and specifically the Na+ content in leaves. Overall, those individuals with presence of endophytes showed higher ecophysiological performance. In addition, the presence of fungal endophytes was correlated with a higher regulation of ion homeostasis by enhanced expression of NHX1 gene. Our results suggest that presence of fungal endophytes could minimize the negative effect of salt by improving osmotic tolerance through ecophysiological and molecular mechanisms. Thus, root-endophytes might be a successful biotechnological tool to maintain high levels of ecophysiological performance and productivity in zones under osmotic stress, acting as potential solution to maintain the global food security.\n\nHighlightWe showed that functional symbiosis improve the physiological performance and yield in crops subjected to salinity, by biochemical and molecular mechanisms. Thus, here we pointed a successful strategy to meet the future challenges for help to maintain the food security.

plant biology

The whale shark genome reveals how genomic and physiological properties scale with body size

The endangered whale shark (Rhincodon typus) is the largest fish on Earth and is a long-lived member of the ancient Elasmobranchii clade. To characterize the relationship between genome features and biological traits, we sequenced and assembled the genome of the whale shark and compared its genomic and physiological features to those of 81 animals and yeast. We examined scaling relationships between body size, temperature, metabolic rates, and genomic features and found both general correlations across the animal kingdom and features specific to the whale shark genome. Among animals, increased lifespan is positively correlated to body size and metabolic rate. Several genomic features also significantly correlated with body size, including intron and gene length. Our large-scale comparative genomic analysis uncovered general features of metazoan genome architecture: GC content and codon adaptation index are negatively correlated, and neural connectivity genes are longer than average genes in most genomes. Focusing on the whale shark genome, we identified multiple features that significantly correlate with lifespan. Among these were very long gene length, due to large introns highly enriched in repetitive elements such as CR1-like LINEs, and considerably longer neural genes of several types, including connectivity, activity, and neurodegeneration genes. The whale sharks genome had an expansion of gene families related to fatty acid metabolism and neurogenesis, with the slowest evolutionary rate observed in vertebrates to date. Our comparative genomics approach uncovered multiple genetic features associated with body size, metabolic rate, and lifespan, and showed that the whale shark is a promising model for studies of neural architecture and lifespan.

genomics

Stressed mothers, tolerant daughters: a case study about the physiological responses and growth of sugarcane plants under water deficit

Drought stress can imprint marks in plants after a previous exposure, leading to a permissive state that facilitates a more effective response to subsequent stress events. Such stress imprints would benefit plants obtained from progenitors previously exposed to drought. Herein, our hypothesis was that daughter plants obtained from mother plants previously exposed to water deficit will perform better under water deficit as compared to those obtained from mothers that did not face stressful conditions. Sugarcane mother plants were grown under well-hydrated conditions or subjected to three cycles of water deficit by water withholding. Then, daughter plants produced through vegetative propagation were subjected to water deficit. Leaf gas exchange was reduced under water deficit and daughters from mothers that experienced water deficit presented a faster recovery of CO2 assimilation and higher instantaneous carboxylation efficiency after rehydration as compared to daughters from mothers that did not face water deficit. Plants obtained from mother plants that faced water deficit showed the highest leaf proline concentration under water deficit as well as higher leaf H2O 2 concentration and leaf ascorbate peroxidase activity regardless of water regime. Under well-watered conditions, daughters from mothers that faced stressful conditions presented higher root H2O2 concentration and root catalase activity than ones from mothers that did not experience water shortage. Such physiological changes were associated with improvements in leaf area and shoot and root dry matter accumulation in daughters from stressed mothers. Our results suggest that root H2O2 concentration is a chemical signal associated with stress memory and improved sugarcane growth. Such findings bring a new perspective to sugarcane production systems, in which stress memory can be explored for improving drought tolerance in rainfed areas.

plant biology

Different factors shape pigmentation and other physiological trait variation within and between species in the Drosophila americana group.

BackgroundDisentangling the selective factors shaping adaptive trait variation is an important but challenging task. Many studies--especially in Drosophila--have documented trait variation along latitudinal or altitudinal clines, but frequently lack resolution about specific environmental gradients that could be causal selective agents, and often do not investigate covariation between traits simultaneously. Here we examined variation in multiple macroecological factors across geographic space and their associations with variation in three physiological traits (desiccation resistance, UV resistance, and pigmentation) at both population and species scales, to address the role of abiotic environment in shaping trait variation.\n\nResultsUsing environmental data from collection locations of three North American Drosophila species--D. americana americana, D. americana texana and D. novamexicana--we identified two primary axes of macroecological variation; these differentiated species habitats and were strongly loaded for precipitation and moisture variables. In nine focal populations (three per species) assayed for each trait, we detected significant species-level variation for both desiccation resistance and pigmentation, but not for UV resistance. Species-level trait variation was consistent with differential natural selection imposed by variation in habitat water availability, although patterns of variation differed between desiccation resistance and pigmentation, and we found little evidence for pleiotropy between traits.\n\nConclusionsOur multi-faceted approach enabled us to identify potential agents of natural selection and examine how they might influence the evolution of multiple traits at different evolutionary scales. Our findings highlight that environmental factors influence functional trait variation in ways that can be complex, and point to the importance of studies that examine these relationships at both population- and species-levels.

evolutionary biology

The Unreasonable Effectiveness of Cell Types in Describing Neuronal Physiological Features

Single-cell RNA sequencing (scRNA-seq) captures detailed gene expression profiles at scale, while patch-clamp recordings measure intrinsic neuronal electrophysiological properties. Modeling the relations between these two modalities remains a challenge. Here, we compare how well electrophysiological features can be predicted by traditional transcriptomic cell type classification, representations derived from a foundational model (scGPT) pretrained on large-scale scRNA-seq datasets, ion channel-coding genes, and highly variable genes. Using paired transcriptomic and electrophysiological patch-sequencing data from 495 human neurons from neurosurgical tissue, we find that cluster-level cell type representations consistently outperform highly variable gene selection, ion channel gene selection, and context-enriched scGPT embeddings. Notably, performance varies across model architectures and initializations, and the best results are obtained by combining the outputs of separate cell type and scGPT-based models. Together, these findings suggest that traditional discrete cellular classification is highly effective in predicting physiological features. For maximum performance it can be complemented by pretrained transformer models.

neuroscience

Understanding the physiological alterations of Vibrio cholerae upon exposure to L-ascorbic acid

The scourge of cholera remains a major global public health threat. It affects up to 4 million people worldwide and causes tens of thousands of deaths each year. The disease is experiencing a concerning resurgence in many parts of Africa, the Middle East, and Asia. To effectively tackle cholera and circumvent rising antimicrobial resistance, targeted biological and preventive approaches, complementing traditional rehydration, are urgently needed. In this regard, our group has demonstrated the efficacy of L-ascorbic acid in controlling the growth and pathogenesis of Vibrio cholerae in vitro. The present work further provides a mechanistic elucidation of the L-ascorbic acid-mediated physiological changes in V. cholerae and also bolsters such a non-antibiotic approach to control cholera.

microbiology

Physiological and anatomical leaf acclimation of understory trees subjected to a through-fall precipitation exclusion in a temperate rain forest in southern South America.

Water input is a key component of the ecosystems. Water defines the functionality, composition, and structure of biomes; therefore, any change in its availability would have an impact on ecosystem features. Moreover, a change in the water balance of an ecosystem affects its persistence as well as biochemical cycles, such as the carbon and nitrogen. Forests are ecosystems structured using high amounts of water. Thus, trees, the oldest living plants, are the prime species in these ecosystems and are the main managers of this abiotic element. Trees uptake water from the soil, store it in their biomass, and exchange it with the environment through leaf stomata. They also intercepted rainfall and fog with their canopies. All of this water is also transmitted to the entire biological diversity that inhabits these ecosystems. Any change in water input affects the web described above. The ability of trees to modify their anatomy or processes, that is, to acclimate to novel climates, is of great advantage in maintaining the characteristics of ecosystems. In this study, we took advantage of a precipitation exclusion experiment to reveal the acclimation of shade-tolerant understory trees, which will be the main component of a cold temperate rainforest in the future. We evaluated different anatomical and physiological leaf traits involved in the use of water by these species. We hypothesized that, as observed in similar experiments, species would adopt more conservative water-use strategies by adjusting their functional traits accordingly. Contrary to our hypotheses, we found that understory tree species inhabiting this temperate ecosystem will not become more conservative when using water. In minimal, but significant differences, most of the studied species displayed traits, in the precipitation exclusion treatment, that were demonstrated to be water spender, rather than conservative. We attributed these contrasting changes to root metabolism alleviation due to the flooded soils of Chiloe inhabited by these forests.

ecology