RESUMO
Temperature is a global factor that affects the performance of all intracellular networks. Robustness against temperature variations is thus expected to be an essential network property, particularly in organisms without inherent temperature control. Here, we combine experimental analyses with computational modeling to investigate thermal robustness of signaling in chemotaxis of Escherichia coli, a relatively simple and well-established model for systems biology. We show that steady-state and kinetic pathway parameters that are essential for chemotactic performance are indeed temperature-compensated in the entire physiological range. Thermal robustness of steady-state pathway output is ensured at several levels by mutual compensation of temperature effects on activities of individual pathway components. Moreover, the effect of temperature on adaptation kinetics is counterbalanced by preprogrammed temperature dependence of enzyme synthesis and stability to achieve nearly optimal performance at the growth temperature. Similar compensatory mechanisms are expected to ensure thermal robustness in other systems.
Assuntos
Quimiotaxia , Escherichia coli/fisiologia , Transdução de Sinais , Adaptação Fisiológica , Escherichia coli/enzimologia , Transferência Ressonante de Energia de Fluorescência , Cinética , Metilação , Monoéster Fosfórico Hidrolases/metabolismo , Fosfotransferases/metabolismo , TemperaturaRESUMO
Caenorhabditis elegans is capable of learning and remembering behaviorally relevant cues such as smells, tastes, and temperature. This is an example of associative learning, a process in which behavior is modified by making associations between various stimuli. Since the mathematical theory of conditioning does not account for some of its salient aspects, such as spontaneous recovery of extinguished associations, accurate modeling of behavior of real animals during conditioning has turned out difficult. Here, we do this in the context of the dynamics of the thermal preference of C. elegans. We quantify C. elegans thermotaxis in response to various conditioning temperatures, starvation durations, and genetic perturbations using a high-resolution microfluidic droplet assay. We model these data comprehensively, within a biologically interpretable, multi-modal framework. We find that the strength of the thermal preference is composed of two independent, genetically separable contributions and requires a model with at least four dynamical variables. One pathway positively associates the experienced temperature independently of food and the other negatively associates with the temperature when food is absent. The multidimensional structure of the association strength provides an explanation for the apparent classical temperature-food association of C. elegans thermal preference and a number of longstanding questions in animal learning, including spontaneous recovery, asymmetric response to appetitive vs. aversive cues, latent inhibition, and generalization among similar cues.
Assuntos
Proteínas de Caenorhabditis elegans , Caenorhabditis elegans , Animais , Caenorhabditis elegans/metabolismo , Comportamento Animal/fisiologia , Aprendizagem , Temperatura , Proteínas de Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/metabolismoRESUMO
The roundworm Caenorhabditis elegans exhibits robust escape behavior in response to rapidly rising temperature. The behavior lasts for a few seconds, shows history dependence, involves both sensory and motor systems, and is too complicated to model mechanistically using currently available knowledge. Instead we model the process phenomenologically, and we use the Sir Isaac dynamical inference platform to infer the model in a fully automated fashion directly from experimental data. The inferred model requires incorporation of an unobserved dynamical variable and is biologically interpretable. The model makes accurate predictions about the dynamics of the worm behavior, and it can be used to characterize the functional logic of the dynamical system underlying the escape response. This work illustrates the power of modern artificial intelligence to aid in discovery of accurate and interpretable models of complex natural systems.
Assuntos
Caenorhabditis elegans/fisiologia , Reação de Fuga/fisiologia , Temperatura Alta , Modelos Biológicos , AnimaisRESUMO
BACKGROUND: Animal responses to thermal stimuli involve intricate contributions of genetics, neurobiology and physiology, with temperature variation providing a pervasive environmental factor for natural selection. Thermal behavior thus exemplifies a dynamic trait that requires non-trivial phenotypic summaries to appropriately capture the trait in response to a changing environment. To characterize the deterministic and plastic components of thermal responses, we developed a novel micro-droplet assay of nematode behavior that permits information-dense summaries of dynamic behavioral phenotypes as reaction norms in response to increasing temperature (thermal tolerance curves, TTC). RESULTS: We found that C. elegans TTCs shift predictably with rearing conditions and developmental stage, with significant differences between distinct wildtype genetic backgrounds. Moreover, after screening TTCs for 58 C. elegans genetic mutant strains, we determined that genes affecting thermosensation, including cmk-1 and tax-4, potentially play important roles in the behavioral control of locomotion at high temperature, implicating neural decision-making in TTC shape rather than just generalized physiological limits. However, expression of the transient receptor potential ion channel TRPA-1 in the nervous system is not sufficient to rescue rearing-dependent plasticity in TTCs conferred by normal expression of this gene, indicating instead a role for intestinal signaling involving TRPA-1 in the adaptive plasticity of thermal performance. CONCLUSIONS: These results implicate nervous system and non-nervous system contributions to behavior, in addition to basic cellular physiology, as key mediators of evolutionary responses to selection from temperature variation in nature.
Assuntos
Adaptação Fisiológica/fisiologia , Proteínas de Caenorhabditis elegans/fisiologia , Caenorhabditis elegans , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/fisiologia , Canais Iônicos/fisiologia , Locomoção/fisiologia , Canal de Cátion TRPA1/fisiologia , Sensação Térmica/fisiologia , Adaptação Fisiológica/genética , Animais , Proteínas de Caenorhabditis elegans/biossíntese , Proteínas de Caenorhabditis elegans/genética , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/genética , Temperatura Alta , Canais Iônicos/genética , Estágios do Ciclo de Vida/fisiologia , Mutação , Sistema Nervoso/metabolismo , Canal de Cátion TRPA1/biossínteseRESUMO
A goal of many sensorimotor studies is to quantify the stimulus-behavioral response relation for specific organisms and specific sensory stimuli. This is especially important to do in the context of painful stimuli since most animals in these studies cannot easily communicate to us their perceived levels of such noxious stimuli. Thus progress on studies of nociception and pain-like responses in animal models depends crucially on our ability to quantitatively and objectively infer the sensed levels of these stimuli from animal behaviors. Here we develop a quantitative model to infer the perceived level of heat stimulus from the stereotyped escape response of individual nematodes Caenorhabditis elegans stimulated by an IR laser. The model provides a method for quantification of analgesic-like effects of chemical stimuli or genetic mutations in C. elegans. We test ibuprofen-treated worms and a TRPV (transient receptor potential) mutant, and we show that the perception of heat stimuli for the ibuprofen treated worms is lower than the wild-type. At the same time, our model shows that the mutant changes the worm's behavior beyond affecting the thermal sensory system. Finally, we determine the stimulus level that best distinguishes the analgesic-like effects and the minimum number of worms that allow for a statistically significant identification of these effects.
Assuntos
Caenorhabditis elegans/fisiologia , Reação de Fuga/fisiologia , Resposta ao Choque Térmico/fisiologia , Modelos Biológicos , Percepção da Dor/fisiologia , Estimulação Física/métodos , Animais , Simulação por Computador , Temperatura Alta , Medição da Dor/métodos , Comportamento Estereotipado/fisiologiaRESUMO
Understanding the structural complexity and the main drivers of animal search behaviour is pivotal to foraging ecology. Yet, the role of uncertainty as a generative mechanism of movement patterns is poorly understood. Novel insights from search theory suggest that organisms should collect and assess new information from the environment by producing complex exploratory strategies. Based on an extension of the first passage time theory, and using simple equations and simulations, we unveil the elementary heuristics behind search behaviour. In particular, we show that normal diffusion is not enough for determining optimal exploratory behaviour but anomalous diffusion is required. Searching organisms go through two critical sequential phases (approach and detection) and experience fundamental search tradeoffs that may limit their encounter rates. Using experimental data, we show that biological search includes elements not fully considered in contemporary physical search theory. In particular, the need to consider search movement as a non-stationary process that brings the organism from one informational state to another. For example, the transition from remaining in an area to departing from it may occur through an exploratory state where cognitive search is challenged. Therefore, a more comprehensive view of foraging ecology requires including current perspectives about movement under uncertainty.
Assuntos
Comportamento Alimentar/fisiologia , Modelos Biológicos , Atividade Motora/fisiologia , Animais , Caenorhabditis elegans/fisiologia , Simulação por Computador , Fatores de TempoRESUMO
Undulatory motion is common to many creatures across many scales, from sperm to snakes. These organisms must push off against their external environment, such as a viscous medium, grains of sand, or a high-friction surface; additionally they must work to bend their own body. A full understanding of undulatory motion, and locomotion in general, requires the characterization of the material properties of the animal itself. The material properties of the model organism Caenorhabditis elegans were studied with a micromechanical experiment used to carry out a three-point bending measurement of the worm. Worms at various developmental stages (including dauer) were measured and different positions along the worm were probed. From these experiments we calculated the viscoelastic properties of the worm, including the effective spring constant and damping coefficient of bending. C. elegans moves by propagating sinusoidal waves along its body. Whereas previous viscoelastic approaches to describe the undulatory motion have used a Kelvin-Voigt model, where the elastic and viscous components are connected in parallel, our measurements show that the Maxwell model, where the elastic and viscous components are in series, is more appropriate. The viscous component of the worm was shown to be consistent with a non-Newtonian, shear-thinning fluid. We find that as the worm matures it is well described as a self-similar elastic object with a shear-thinning damping term and a stiffness that becomes smaller as one approaches the tail.
Assuntos
Caenorhabditis elegans/fisiologia , Locomoção/fisiologia , Modelos Biológicos , Animais , ElasticidadeRESUMO
The viscoelastic material properties of the model organism C. elegans were probed with a micropipette deflection technique and modelled with the standard linear solid model. Dynamic relaxation measurements were performed on the millimetric nematode to investigate its viscous characteristics in detail. We show that the internal properties of C. elegans can not be fully described by a simple Newtonian fluid. Instead, a power-law fluid model was implemented and shown to be in excellent agreement with experimental results. The nematode exhibits shear thinning properties and its complex fluid characteristics were quantified. The bending-rate dependence of the internal damping coefficient of C. elegans could affect its gait modulation in different external environments.
Assuntos
Caenorhabditis elegans/fisiologia , Elasticidade , Hidrodinâmica , Modelos Biológicos , Viscosidade , AnimaisRESUMO
With a simple and versatile microcantilever-based force measurement technique, we have probed the drag forces involved in Caenorhabditis elegans locomotion. As a worm crawls on an agar surface, we found that substrate viscoelasticity introduces nonlinearities in the force-velocity relationships, yielding nonconstant drag coefficients that are not captured by original resistive force theory. A major contributing factor to these nonlinearities is the formation of a shallow groove on the agar surface. We measured both the adhesion forces that cause the worm's body to settle into the agar and the resulting dynamics of groove formation. Furthermore, we quantified the locomotive forces produced by C. elegans undulatory motions on a wet viscoelastic agar surface. We show that an extension of resistive force theory is able to use the dynamics of a nematode's body shape along with the measured drag coefficients to predict the forces generated by a crawling nematode.
Assuntos
Caenorhabditis elegans/fisiologia , Locomoção , Animais , Fenômenos BiomecânicosRESUMO
The tangling of two tethered microswimming worms serving as the ends of "active strings" is investigated experimentally and modeled analytically. C. elegans nematodes of similar size are caught by their tails using micropipettes and left to swim and interact at different separations over long times. The worms are found to tangle in a reproducible and statistically predictable manner, which is modeled based on the relative motion of the worm heads. Our results provide insight into the intricate tangling interactions present in active biological systems.
Assuntos
Caenorhabditis elegans/fisiologia , Modelos Biológicos , Natação/fisiologia , AnimaisRESUMO
Animal behaviors often are decomposable into discrete, stereotyped elements, well separated in time. In one model, such behaviors are triggered by specific commands; in the extreme case, the discreteness of behavior is traced to the discreteness of action potentials in the individual command neurons. Here, we use the crawling behavior of the nematode Caenorhabditis elegans to demonstrate the opposite view, in which discreteness, stereotypy, and long timescales emerge from the collective dynamics of the behavior itself. In previous work, we found that as C. elegans crawls, its body moves through a "shape space" in which four dimensions capture approximately 95% of the variance in body shape. Here we show that stochastic dynamics within this shape space predicts transitions between attractors corresponding to abrupt reversals in crawling direction. With no free parameters, our inferred stochastic dynamical system generates reversal timescales and stereotyped trajectories in close agreement with experimental observations. We use the stochastic dynamics to show that the noise amplitude decreases systematically with increasing time away from food, resulting in longer bouts of forward crawling and suggesting that worms can use noise to modify their locomotory behavior.
Assuntos
Comportamento Animal/fisiologia , Caenorhabditis elegans/fisiologia , Locomoção/fisiologia , Modelos Biológicos , Comportamento Estereotipado/fisiologia , Animais , Microscopia de Vídeo , Fatores de TempoRESUMO
BACKGROUND: Nociception evokes a rapid withdrawal behavior designed to protect the animal from potential danger. C. elegans performs a reflexive reversal or forward locomotory response when presented with noxious stimuli at the head or tail, respectively. Here, we have developed an assay with precise spatial and temporal control of an infrared laser stimulus that targets one-fifth of the worm's body and quantifies multiple aspects of the worm's escape response. RESULTS: When stimulated at the head, we found that the escape response can be elicited by changes in temperature as small as a fraction of a degree Celsius, and that aspects of the escape behavior such as the response latency and the escape direction change advantageously as the amplitude of the noxious stimulus increases. We have mapped the behavioral receptive field of thermal nociception along the entire body of the worm, and show a midbody avoidance behavior distinct from the head and tail responses. At the midbody, the worm is sensitive to a change in the stimulus location as small as 80 µm. This midbody response is probabilistic, producing either a backward, forward or pause state after the stimulus. The distribution of these states shifts from reverse-biased to forward-biased as the location of the stimulus moves from the middle towards the anterior or posterior of the worm, respectively. We identified PVD as the thermal nociceptor for the midbody response using calcium imaging, genetic ablation and laser ablation. Analyses of mutants suggest the possibility that TRPV channels and glutamate are involved in facilitating the midbody noxious response. CONCLUSION: Through high resolution quantitative behavioral analysis, we have comprehensively characterized the C. elegans escape response to noxious thermal stimuli applied along its body, and found a novel midbody response. We further identified the nociceptor PVD as required to sense noxious heat at the midbody and can spatially differentiate localized thermal stimuli.
Assuntos
Reação de Fuga/fisiologia , Nociceptividade/fisiologia , Nociceptores/fisiologia , Animais , Caenorhabditis elegans , Proteínas de Caenorhabditis elegans/genética , Temperatura Alta , Proteínas com Homeodomínio LIM/genética , Fatores de Transcrição/genéticaRESUMO
Temperature-dependent behaviours in Caenorhabditis elegans, such as thermotaxis and isothermal tracking, are complex behavioural responses that integrate sensation, foraging and learning, and have driven investigations to discover many essential genetic and neural pathways. The ease of manipulation of the Caenorhabditis model system also has encouraged its application to comparative analyses of phenotypic evolution, particularly contrasts of the classic model C. elegans with C. briggsae. And yet few studies have investigated natural genetic variation in behaviour in any nematode. Here we measure thermotaxis and isothermal tracking behaviour in genetically distinct strains of C. briggsae, further motivated by the latitudinal differentiation in C. briggsae that is associated with temperature-dependent fitness differences in this species. We demonstrate that C. briggsae performs thermotaxis and isothermal tracking largely similar to that of C. elegans, with a tendency to prefer its rearing temperature. Comparisons of these behaviours among strains reveal substantial heritable natural variation within each species that corresponds to three general patterns of behavioural response. However, intraspecific genetic differences in thermal behaviour often exceed interspecific differences. These patterns of temperature-dependent behaviour motivate further development of C. briggsae as a model system for dissecting the genetic underpinnings of complex behavioural traits.
Assuntos
Caenorhabditis/fisiologia , Fenótipo , Animais , Regulação da Temperatura Corporal , Caenorhabditis/genética , Caenorhabditis elegans/genética , Caenorhabditis elegans/fisiologia , Característica Quantitativa Herdável , Especificidade da Espécie , TemperaturaRESUMO
BACKGROUND: Responding to noxious stimuli by invoking an appropriate escape response is critical for survival of an organism. The sensations of small and large changes in temperature in most organisms have been studied separately in the context of thermotaxis and nociception, respectively. Here we use the nematode C. elegans to address the neurogenetic basis of responses to thermal stimuli over a broad range of intensities. RESULTS: C. elegans responds to aversive temperature by eliciting a stereotypical behavioral sequence. Upon sensation of the noxious stimulus, it moves backwards, turns and resumes forward movement in a new direction. In order to study the response of C. elegans to a broad range of noxious thermal stimuli, we developed a novel assay that allows simultaneous characterization of multiple aspects of escape behavior elicited by thermal pulses of increasing amplitudes. We exposed the laboratory strain N2, as well as 47 strains with defects in various aspects of nervous system function, to thermal pulses ranging from ΔT = 0.4°C to 9.1°C and recorded the resulting behavioral profiles. CONCLUSIONS: Through analysis of the multidimensional behavioral profiles, we found that the combinations of molecules shaping avoidance responses to a given thermal pulse are unique. At different intensities of aversive thermal stimuli, these distinct combinations of molecules converge onto qualitatively similar stereotyped behavioral sequences.
Assuntos
Comportamento Animal/fisiologia , Caenorhabditis elegans/fisiologia , Sensação Térmica/fisiologia , Animais , Aprendizagem da Esquiva/fisiologia , Proteínas de Caenorhabditis elegans/metabolismo , Análise por Conglomerados , Reação de Fuga/fisiologia , Glutamatos/metabolismo , Temperatura Alta , Movimento , Neuropeptídeos/metabolismo , Neurotransmissores/metabolismo , Fenótipo , Estimulação Física , Canais de Potássio/metabolismo , Análise de Componente Principal , Transmissão Sináptica/fisiologiaRESUMO
Swimming Escherichia coli responds to changes in temperature by modifying its motor behavior. Previous studies using populations of cells have shown that E. coli accumulate in spatial thermal gradients, but these experiments did not cleanly separate thermal responses from chemotactic responses. Here we have isolated the thermal response by studying the behavior of single, tethered cells. The motor output of cells grown at 33 degrees C was measured at constant temperature, from 10 degrees to 40 degrees C, and in response to small, impulsive increases in temperature, from 23 degrees to 43 degrees C. The thermal impulse response at temperatures < 31 degrees C is similar to the chemotactic impulse response: Both follow a similar time course, share the same directionality, and show biphasic characteristics. At temperatures > 31 degrees C, some cells show an inverted response, switching from warm- to cold-seeking behavior. The fraction of inverted responses increases nonlinearly with temperature, switching steeply at the preferred temperature of 37 degrees C.
Assuntos
Fenômenos Fisiológicos Bacterianos , Escherichia coli/fisiologia , Temperatura , Flagelos/fisiologia , Sensação TérmicaRESUMO
BACKGROUND: Parkinson's disease is a disabling neurodegenerative movement disorder characterized by dopaminergic neuron loss induced by α-synuclein oligomers. There is an urgent need for disease-modifying therapies for Parkinson's disease, but drug discovery is challenged by lack of in vivo models that recapitulate early stages of neurodegeneration. Invertebrate organisms, such as the nematode worm Caenorhabditis elegans, provide in vivo models of human disease processes that can be instrumental for initial pharmacological studies. METHODS: To identify early motor impairment of animals expressing α-synuclein in dopaminergic neurons, we first used a custom-built tracking microscope that captures locomotion of single C. elegans with high spatial and temporal resolution. Next, we devised a method for semi-automated and blinded quantification of motor impairment for a population of simultaneously recorded animals with multi-worm tracking and custom image processing. We then used genetic and pharmacological methods to define the features of early motor dysfunction of α-synuclein-expressing C. elegans. Finally, we applied the C. elegans model to a drug repurposing screen by combining it with an artificial intelligence platform and cell culture system to identify small molecules that inhibit α-synuclein oligomers. Screen hits were validated using in vitro and in vivo mammalian models. RESULTS: We found a previously undescribed motor phenotype in transgenic α-synuclein C. elegans that correlates with mutant or wild-type α-synuclein protein levels and results from dopaminergic neuron dysfunction, but precedes neuronal loss. Together with artificial intelligence-driven in silico and in vitro screening, this C. elegans model identified five compounds that reduced motor dysfunction induced by α-synuclein. Three of these compounds also decreased α-synuclein oligomers in mammalian neurons, including rifabutin which has not been previously investigated for Parkinson's disease. We found that treatment with rifabutin reduced nigrostriatal dopaminergic neurodegeneration due to α-synuclein in a rat model. CONCLUSIONS: We identified a C. elegans locomotor abnormality due to dopaminergic neuron dysfunction that models early α-synuclein-mediated neurodegeneration. Our innovative approach applying this in vivo model to a multi-step drug repurposing screen, with artificial intelligence-driven in silico and in vitro methods, resulted in the discovery of at least one drug that may be repurposed as a disease-modifying therapy for Parkinson's disease.
Assuntos
Transtornos Motores , alfa-Sinucleína , Animais , Inteligência Artificial , Caenorhabditis elegans/metabolismo , Modelos Animais de Doenças , Dopamina/metabolismo , Neurônios Dopaminérgicos/metabolismo , Mamíferos/metabolismo , Transtornos Motores/metabolismo , Ratos , alfa-Sinucleína/metabolismoRESUMO
The effects of the nanocrystal size on the emission spectra and decay rates of upconverting hexagonal NaYF(4):Yb,Er nanocrystals are investigated. The influence of nanocrystal size is represented in terms of the surface area/volume ratio (SA/Vol). Our results show that a small nanocrystal size, or large SA/Vol ratio increases the decay rate, in particular, the green luminescence decay rate varies linearly with the SA/Vol ratio.
RESUMO
We have developed a powerful experimental framework that combines competitive selection and microarray-based genetic footprinting to comprehensively reveal the genetic basis of bacterial behaviors. Application of this method to Escherichia coli motility identifies 95% of the known flagellar and chemotaxis genes, and reveals three dozen novel loci that, to varying degrees and through diverse mechanisms, affect motility. To probe the network context in which these genes function, we developed a method that uncovers genome-wide epistatic interactions through comprehensive analyses of double-mutant phenotypes. This allows us to place the novel genes within the context of signaling and regulatory networks, including the Rcs phosphorelay pathway and the cyclic di-GMP second-messenger system. This unifying framework enables sensitive and comprehensive genetic characterization of complex behaviors across the microbial biosphere.
Assuntos
Bacillus subtilis/fisiologia , Quimiotaxia , Escherichia coli/fisiologia , Bacillus subtilis/genética , Epistasia Genética , Escherichia coli/genética , Genoma Bacteriano , Sistemas do Segundo Mensageiro , Transdução de SinaisRESUMO
The ability to avoid harmful or potentially harmful stimuli can help an organism escape predators and injury, and certain avoidance mechanisms are conserved across the animal kingdom. However, how the need to avoid an imminent threat is balanced with current behavior and modified by past experience is not well understood. In this work we focused on rapidly increasing temperature, a signal that triggers an escape response in a variety of animals, including the nematode Caenorhabditis elegans. We have developed a noxious thermal response assay using an infrared laser that can be automatically controlled and targeted in order to investigate how C. elegans responds to noxious heat over long timescales and to repeated stimuli in various behavioral and sensory contexts. High-content phenotyping of behavior in individual animals revealed that the C. elegans escape response is multidimensional, with some features that extend for several minutes, and can be modulated by (i) stimulus amplitude; (ii) other sensory inputs, such as food context; (iii) long and short-term thermal experience; and (iv) the animal's current behavioral state.
Assuntos
Aprendizagem da Esquiva/fisiologia , Comportamento Animal/fisiologia , Caenorhabditis elegans/fisiologia , Reação de Fuga/fisiologia , Temperatura Alta , Células Receptoras Sensoriais/metabolismo , Sensação Térmica/fisiologia , Animais , Fenótipo , Células Receptoras Sensoriais/citologiaRESUMO
A major challenge in analyzing animal behavior is to discover some underlying simplicity in complex motor actions. Here, we show that the space of shapes adopted by the nematode Caenorhabditis elegans is low dimensional, with just four dimensions accounting for 95% of the shape variance. These dimensions provide a quantitative description of worm behavior, and we partially reconstruct "equations of motion" for the dynamics in this space. These dynamics have multiple attractors, and we find that the worm visits these in a rapid and almost completely deterministic response to weak thermal stimuli. Stimulus-dependent correlations among the different modes suggest that one can generate more reliable behaviors by synchronizing stimuli to the state of the worm in shape space. We confirm this prediction, effectively "steering" the worm in real time.