RESUMO
To fully understand animal transcription networks, it is essential to accurately measure the spatial and temporal expression patterns of transcription factors and their targets. We describe a registration technique that takes image-based data from hundreds of Drosophila blastoderm embryos, each costained for a reference gene and one of a set of genes of interest, and builds a model VirtualEmbryo. This model captures in a common framework the average expression patterns for many genes in spite of significant variation in morphology and expression between individual embryos. We establish the method's accuracy by showing that relationships between a pair of genes' expression inferred from the model are nearly identical to those measured in embryos costained for the pair. We present a VirtualEmbryo containing data for 95 genes at six time cohorts. We show that known gene-regulatory interactions can be automatically recovered from this data set and predict hundreds of new interactions.
Assuntos
Drosophila melanogaster/genética , Redes Reguladoras de Genes , Modelos Genéticos , Animais , Blastoderma , Drosophila melanogaster/metabolismo , Embrião não Mamífero/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Genes de InsetosRESUMO
Taxonomic resolution is a major challenge in palynology, largely limiting the ecological and evolutionary interpretations possible with deep-time fossil pollen data. We present an approach for fossil pollen analysis that uses optical superresolution microscopy and machine learning to create a quantitative and higher throughput workflow for producing palynological identifications and hypotheses of biological affinity. We developed three convolutional neural network (CNN) classification models: maximum projection (MPM), multislice (MSM), and fused (FM). We trained the models on the pollen of 16 genera of the legume tribe Amherstieae, and then used these models to constrain the biological classifications of 48 fossil Striatopollis specimens from the Paleocene, Eocene, and Miocene of western Africa and northern South America. All models achieved average accuracies of 83 to 90% in the classification of the extant genera, and the majority of fossil identifications (86%) showed consensus among at least two of the three models. Our fossil identifications support the paleobiogeographic hypothesis that Amherstieae originated in Paleocene Africa and dispersed to South America during the Paleocene-Eocene Thermal Maximum (56 Ma). They also raise the possibility that at least three Amherstieae genera (Crudia, Berlinia, and Anthonotha) may have diverged earlier in the Cenozoic than predicted by molecular phylogenies.
Assuntos
Fósseis , Microscopia/métodos , Redes Neurais de Computação , Filogenia , Pólen/classificação , África , África Ocidental , Aprendizado de Máquina , Filogeografia , América do SulRESUMO
Study of the neural deficits caused by mismatched binocular vision in early childhood has predominantly focused on circuits in the primary visual cortex (V1). Recent evidence has revealed that neurons in mouse dorsolateral geniculate nucleus (dLGN) can undergo rapid ocular dominance plasticity following monocular deprivation (MD). It remains unclear, however, whether the long-lasting deficits attributed to MD during the critical period originate in the thalamus. Using in vivo two-photon Ca2+ imaging of dLGN afferents in superficial layers of V1 in female and male mice, we demonstrate that 14 d MD during the critical period leads to a chronic loss of binocular dLGN inputs while sparing response strength and spatial acuity. Importantly, MD leads to profoundly mismatched visual tuning properties in remaining binocular dLGN afferents. Furthermore, MD impairs binocular modulation, reducing facilitation of responses of both binocular and monocular dLGN inputs during binocular viewing. As predicted by our findings in thalamic inputs, Ca2+ imaging from V1 neurons revealed spared spatial acuity but impaired binocularity in L4 neurons. V1 L2/3 neurons in contrast displayed deficits in both binocularity and spatial acuity. Our data demonstrate that critical-period MD produces long-lasting disruptions in binocular integration beginning in early binocular circuits in dLGN, whereas spatial acuity deficits first arise from circuits further downstream in V1. Our findings indicate that the development of normal binocular vision and spatial acuity depend upon experience-dependent refinement of distinct stages in the mammalian visual system.SIGNIFICANCE STATEMENT Abnormal binocular vision and reduced acuity are hallmarks of amblyopia, a disorder that affects 2%-5% of the population. It is widely thought that the neural deficits underlying amblyopia begin in the circuits of primary visual cortex. Using in vivo two-photon calcium imaging of thalamocortical axons in mice, we show that depriving one eye of input during a critical period in development chronically impairs binocular integration in thalamic inputs to primary visual cortex. In contrast, visual acuity is spared in thalamic inputs. These findings shed new light on the role for developmental mechanisms in the thalamus in establishing binocular vision and may have critical implications for amblyopia.
Assuntos
Privação Sensorial/fisiologia , Tálamo/crescimento & desenvolvimento , Tálamo/fisiologia , Visão Binocular/fisiologia , Visão Monocular/fisiologia , Visão Ocular/fisiologia , Ambliopia/fisiopatologia , Animais , Mapeamento Encefálico , Feminino , Corpos Geniculados/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Estimulação Luminosa , Percepção Espacial , Acuidade Visual/fisiologia , Córtex Visual/fisiologiaRESUMO
Self-renewing organs often experience a decline in function in the course of aging. It is unclear whether chronological age or external factors control this decline, or whether it is driven by stem cell self-renewal-for example, because cycling cells exhaust their replicative capacity and become senescent. Here we assay the relationship between stem cell cycling and senescence in the Caenorhabditis elegans reproductive system, defining this senescence as the progressive decline in "reproductive capacity," i.e. in the number of progeny that can be produced until cessation of reproduction. We show that stem cell cycling diminishes remaining reproductive capacity, at least in part through the DNA damage response. Paradoxically, gonads kept under conditions that preclude reproduction keep cycling and producing cells that undergo apoptosis or are laid as unfertilized gametes, thus squandering reproductive capacity. We show that continued activity is in fact beneficial inasmuch as gonads that are active when reproduction is initiated have more sustained early progeny production. Intriguingly, continued cycling is intermittent-gonads switch between active and dormant states-and in all likelihood stochastic. Other organs face tradeoffs whereby stem cell cycling has the beneficial effect of providing freshly-differentiated cells and the detrimental effect of increasing the likelihood of cancer or senescence; stochastic stem cell cycling may allow for a subset of cells to preserve proliferative potential in old age, which may implement a strategy to deal with uncertainty as to the total amount of proliferation to be undergone over an organism's lifespan.
Assuntos
Envelhecimento/fisiologia , Caenorhabditis elegans/fisiologia , Autorrenovação Celular/fisiologia , Senescência Celular/fisiologia , Reparo do DNA/genética , Animais , Apoptose/genética , Proteínas de Caenorhabditis elegans/genética , Senescência Celular/genética , Dano ao DNA/genética , Proteínas de Ligação a DNA/genética , Feminino , Pontos de Checagem da Fase M do Ciclo Celular/genética , Ovário/fisiologia , Proteína de Replicação A/genética , Reprodução/fisiologia , Inanição/fisiopatologia , Células-Tronco , Fatores de Transcrição/genéticaRESUMO
In developing embryos, gene regulatory networks drive cells towards discrete terminal fates, a process called canalization. We studied the behavior of the anterior-posterior segmentation network in Drosophila melanogaster embryos by depleting a key maternal input, bicoid (bcd), and measuring gene expression patterns of the network at cellular resolution. This method results in a gene expression atlas containing the levels of mRNA or protein expression of 13 core patterning genes over six time points for every cell of the blastoderm embryo. This is the first cellular resolution dataset of a genetically perturbed Drosophila embryo that captures all cells in 3D. We describe the technical developments required to build this atlas and how the method can be employed and extended by others. We also analyze this novel dataset to characterize the degree and timing of cell fate canalization in the segmentation network. We find that in two layers of this gene regulatory network, following depletion of bcd, individual cells rapidly canalize towards normal cell fates. This result supports the hypothesis that the segmentation network directly canalizes cell fate, rather than an alternative hypothesis whereby cells are initially mis-specified and later eliminated by apoptosis. Our gene expression atlas provides a high resolution picture of a classic perturbation and will enable further computational modeling of canalization and gene regulation in this transcriptional network.
Assuntos
Padronização Corporal/genética , Linhagem da Célula/genética , Bases de Dados Genéticas , Drosophila melanogaster/embriologia , Redes Reguladoras de Genes/genética , Transcriptoma/genética , Animais , Proteínas de Drosophila , Proteínas de Homeodomínio , Hibridização In Situ , Interferência de RNA , Reação em Cadeia da Polimerase em Tempo Real , Transativadores/deficiênciaRESUMO
BACKGROUND: Analysis of single cells in their native environment is a powerful method to address key questions in developmental systems biology. Confocal microscopy imaging of intact tissues, followed by automatic image segmentation, provides a means to conduct cytometric studies while at the same time preserving crucial information about the spatial organization of the tissue and morphological features of the cells. This technique is rapidly evolving but is still not in widespread use among research groups that do not specialize in technique development, perhaps in part for lack of tools that automate repetitive tasks while allowing experts to make the best use of their time in injecting their domain-specific knowledge. RESULTS: Here we focus on a well-established stem cell model system, the C. elegans gonad, as well as on two other model systems widely used to study cell fate specification and morphogenesis: the pre-implantation mouse embryo and the developing mouse olfactory epithelium. We report a pipeline that integrates machine-learning-based cell detection, fast human-in-the-loop curation of these detections, and running of active contours seeded from detections to segment cells. The procedure can be bootstrapped by a small number of manual detections, and outperforms alternative pieces of software we benchmarked on C. elegans gonad datasets. Using cell segmentations to quantify fluorescence contents, we report previously-uncharacterized cell behaviors in the model systems we used. We further show how cell morphological features can be used to identify cell cycle phase; this provides a basis for future tools that will streamline cell cycle experiments by minimizing the need for exogenous cell cycle phase labels. CONCLUSIONS: High-throughput 3D segmentation makes it possible to extract rich information from images that are routinely acquired by biologists, and provides insights - in particular with respect to the cell cycle - that would be difficult to derive otherwise.
Assuntos
Caenorhabditis elegans/crescimento & desenvolvimento , Ensaios de Triagem em Larga Escala , Processamento de Imagem Assistida por Computador/métodos , Imageamento Tridimensional/métodos , Mucosa Olfatória/citologia , Análise de Célula Única/métodos , Software , Algoritmos , Animais , Blastocisto/citologia , Blastocisto/metabolismo , Caenorhabditis elegans/metabolismo , Ciclo Celular/fisiologia , Células Cultivadas , Biologia Computacional/métodos , Feminino , Gônadas/citologia , Gônadas/metabolismo , Humanos , Masculino , Camundongos , Microscopia Confocal/métodos , Mucosa Olfatória/metabolismoRESUMO
BACKGROUND: The endogenous ability to dedifferentiate, re-pattern, and re-differentiate adult cells to repair or replace damaged or missing structures is exclusive to only a few tetrapod species. The Mexican axolotl is one example of these species, having the capacity to regenerate multiple adult structures including their limbs by generating a group of progenitor cells, known as the blastema, which acquire pattern and differentiate into the missing tissues. The formation of a limb regenerate is dependent on cells in the connective tissues that retain memory of their original position in the limb, and use this information to generate the pattern of the missing structure. Observations from recent and historic studies suggest that blastema cells vary in their potential to pattern distal structures during the regeneration process; some cells are plastic and can be reprogrammed to obtain new positional information while others are stable. Our previous studies showed that positional information has temporal and spatial components of variation; early bud (EB) and apical late bud (LB) blastema cells are plastic while basal-LB cells are stable. To identify the potential cellular and molecular basis of this variation, we compared these three cell populations using histological and transcriptional approaches. RESULTS: Histologically, the basal-LB sample showed greater tissue organization than the EB and apical-LB samples. We also observed that cell proliferation was more abundant in EB and apical-LB tissue when compared to basal-LB and mature stump tissue. Lastly, we found that genes associated with cellular differentiation were expressed more highly in the basal-LB samples. CONCLUSIONS: Our results characterize histological and transcriptional differences between EB and apical-LB tissue compared to basal-LB tissue. Combined with our results from a previous study, we hypothesize that the stability of positional information is associated with tissue organization, cell proliferation, and pathways of cellular differentiation.
Assuntos
Ambystoma mexicanum/embriologia , Plasticidade Celular/genética , Extremidades/embriologia , Botões de Extremidades/embriologia , Regeneração/genética , Ambystoma mexicanum/genética , Animais , Diferenciação Celular/genética , Plasticidade Celular/fisiologia , Proliferação de Células/genética , Botões de Extremidades/fisiologia , Regeneração/fisiologia , Transdução de Sinais/genéticaRESUMO
Functional brain imaging is a common tool in monitoring the progression of neurodegenerative and neurological disorders. Identifying functional brain imaging derived features that can accurately detect neurological disease is of primary importance to the medical community. Research in computer vision techniques to identify objects in photographs have reported high accuracies in that domain, but their direct applicability to identifying disease in functional imaging is still under investigation in the medical community. In particular, Serre et al. (: In: IEEE Conference on Computer Vision and Pattern Recognition (CVPR-05). pp 994-1000) introduced a biophysically inspired filtering method emulating visual processing in striate cortex which they applied to perform object recognition in photographs. In this work, the model described by Serre et al. [2005] is extended to three-dimensional volumetric images to perform signal detection in functional brain imaging (PET, SPECT). The filter outputs are used to train both neural network and logistic regression classifiers and tested on two distinct datasets: ADNI Alzheimer's disease 2-deoxy-D-glucose (FDG) PET and National Football League players Tc99m HMPAO SPECT. The filtering pipeline is analyzed to identify which steps are most important for classification accuracy. Our results compare favorably with other published classification results and outperform those of a blinded expert human rater, suggesting the utility of this approach.
Assuntos
Encefalopatias/diagnóstico por imagem , Mapeamento Encefálico/métodos , Redes Neurais de Computação , Reconhecimento Automatizado de Padrão/métodos , Algoritmos , Humanos , Tomografia por Emissão de Pósitrons/métodos , Tomografia Computadorizada de Emissão de Fóton Único/métodosRESUMO
Differences in the level, timing, or location of gene expression can contribute to alternative phenotypes at the molecular and organismal level. Understanding the origins of expression differences is complicated by the fact that organismal morphology and gene regulatory networks could potentially vary even between closely related species. To assess the scope of such changes, we used high-resolution imaging methods to measure mRNA expression in blastoderm embryos of Drosophila yakuba and Drosophila pseudoobscura and assembled these data into cellular resolution atlases, where expression levels for 13 genes in the segmentation network are averaged into species-specific, cellular resolution morphological frameworks. We demonstrate that the blastoderm embryos of these species differ in their morphology in terms of size, shape, and number of nuclei. We present an approach to compare cellular gene expression patterns between species, while accounting for varying embryo morphology, and apply it to our data and an equivalent dataset for Drosophila melanogaster. Our analysis reveals that all individual genes differ quantitatively in their spatio-temporal expression patterns between these species, primarily in terms of their relative position and dynamics. Despite many small quantitative differences, cellular gene expression profiles for the whole set of genes examined are largely similar. This suggests that cell types at this stage of development are conserved, though they can differ in their relative position by up to 3-4 cell widths and in their relative proportion between species by as much as 5-fold. Quantitative differences in the dynamics and relative level of a subset of genes between corresponding cell types may reflect altered regulatory functions between species. Our results emphasize that transcriptional networks can diverge over short evolutionary timescales and that even small changes can lead to distinct output in terms of the placement and number of equivalent cells.
Assuntos
Padronização Corporal/genética , Proteínas de Drosophila/metabolismo , Drosophila/embriologia , Drosophila/genética , Animais , Evolução Biológica , Blastoderma/crescimento & desenvolvimento , Proteínas de Drosophila/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Redes Reguladoras de Genes/genética , Hibridização in Situ Fluorescente , Especificidade da EspécieRESUMO
Taxonomic identification of pollen and spores uses inherently qualitative descriptions of morphology. Consequently, identifications are restricted to categories that can be reliably classified by multiple analysts, resulting in the coarse taxonomic resolution of the pollen and spore record. Grass pollen represents an archetypal example; it is not routinely identified below family level. To address this issue, we developed quantitative morphometric methods to characterize surface ornamentation and classify grass pollen grains. This produces a means of quantifying morphological features that are traditionally described qualitatively. We used scanning electron microscopy to image 240 specimens of pollen from 12 species within the grass family (Poaceae). We classified these species by developing algorithmic features that quantify the size and density of sculptural elements on the pollen surface, and measure the complexity of the ornamentation they form. These features yielded a classification accuracy of 77.5%. In comparison, a texture descriptor based on modelling the statistical distribution of brightness values in image patches yielded a classification accuracy of 85.8%, and seven human subjects achieved accuracies between 68.33 and 81.67%. The algorithmic features we developed directly relate to biologically meaningful features of grass pollen morphology, and could facilitate direct interpretation of unsupervised classification results from fossil material.
Assuntos
Classificação/métodos , Poaceae/anatomia & histologia , Pólen/anatomia & histologia , Fósseis , Microscopia Eletrônica de Varredura , Poaceae/classificação , Pólen/classificaçãoRESUMO
Remodeling of injured sympathetic nerves on the heart after myocardial infarction (MI) contributes to adverse outcomes such as sudden arrhythmic death, yet the underlying structural mechanisms are poorly understood. We sought to examine microstructural changes on the heart after MI and to directly link these changes with electrical dysfunction. We developed a high-resolution pipeline for anatomically precise alignment of electrical maps with structural myofiber and nerve-fiber maps created by customized computer vision algorithms. Using this integrative approach in a mouse model, we identified distinct structure-function correlates to objectively delineate the infarct border zone, a known source of arrhythmias after MI. During tyramine-induced sympathetic nerve activation, we demonstrated regional patterns of altered electrical conduction aligned directly with altered neuroeffector junction distribution, pointing to potential neural substrates for cardiac arrhythmia. This study establishes a synergistic framework for examining structure-function relationships after MI with microscopic precision that has potential to advance understanding of arrhythmogenic mechanisms.
Assuntos
Mapeamento Potencial de Superfície Corporal/métodos , Infarto do Miocárdio/diagnóstico , Miocárdio/patologia , Sistema Nervoso Simpático/diagnóstico por imagem , Potenciais de Ação , Animais , Modelos Animais de Doenças , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Infarto do Miocárdio/fisiopatologia , Sistema Nervoso Simpático/fisiopatologiaRESUMO
The shape of the contour separating two regions strongly influences judgments of which region is "figure" and which is "ground." Convexity and other figure-ground cues are generally assumed to indicate only which region is nearer, but nothing about how much the regions are separated in depth. To determine the depth information conveyed by convexity, we examined natural scenes and found that depth steps across surfaces with convex silhouettes are likely to be larger than steps across surfaces with concave silhouettes. In a psychophysical experiment, we found that humans exploit this correlation. For a given binocular disparity, observers perceived more depth when the near surface's silhouette was convex rather than concave. We estimated the depth distributions observers used in making those judgments: they were similar to the natural-scene distributions. Our findings show that convexity should be reclassified as a metric depth cue. They also suggest that the dichotomy between metric and nonmetric depth cues is false and that the depth information provided many cues should be evaluated with respect to natural-scene statistics. Finally, the findings provide an explanation for why figure-ground cues modulate the responses of disparity-sensitive cells in visual cortex.
Assuntos
Sinais (Psicologia) , Percepção de Profundidade/fisiologia , Área de Dependência-Independência , Modelos Biológicos , Natureza , Reconhecimento Visual de Modelos/fisiologia , Análise de Variância , Humanos , Estimulação Luminosa/métodos , Valor Preditivo dos Testes , Probabilidade , Psicofísica , Disparidade Visual , Visão OcularRESUMO
Disentangling the sources of visual motion in a dynamic scene during self-movement or ego motion is important for autonomous navigation and tracking. In the dynamic image segments of a video frame containing independently moving objects, optic flow relative to the next frame is the sum of the motion fields generated due to camera and object motion. The traditional ego-motion estimation methods assume the scene to be static, and the recent deep learning-based methods do not separate pixel velocities into object- and ego-motion components. We propose a learning-based approach to predict both ego-motion parameters and object-motion field (OMF) from image sequences using a convolutional autoencoder while being robust to variations due to the unconstrained scene depth. This is achieved by: 1) training with continuous ego-motion constraints that allow solving for ego-motion parameters independently of depth and 2) learning a sparsely activated overcomplete ego-motion field (EMF) basis set, which eliminates the irrelevant components in both static and dynamic segments for the task of ego-motion estimation. In order to learn the EMF basis set, we propose a new differentiable sparsity penalty function that approximates the number of nonzero activations in the bottleneck layer of the autoencoder and enforces sparsity more effectively than L1- and L2-norm-based penalties. Unlike the existing direct ego-motion estimation methods, the predicted global EMF can be used to extract OMF directly by comparing it against the optic flow. Compared with the state-of-the-art baselines, the proposed model performs favorably on pixelwise object- and ego-motion estimation tasks when evaluated on real and synthetic data sets of dynamic scenes.
RESUMO
BACKGROUND: Intravenous administration of adeno-associated virus (AAV) can be used as a noninvasive approach to trace neuronal morphology and links. AAV-PHP.S is a variant of AAV9 that effectively transduces the peripheral nervous system. The objective was to label randomly and sparsely enteric plexus in the mouse colon using AAV-PHP.S with a tunable two-component multicolor vector system and digitally trace individual neurons and nerve fibers within microcircuits in three dimensions (3D). METHODS: A vector system including a tetracycline inducer with a tet-responsive element driving three separate fluorophores was packaged in the AAV-PHP.S capsid. The vectors were injected retro-orbitally in mice, and the colon was harvested 3 weeks after. Confocal microscopic images of enteric plexus were digitally segmented and traced in 3D using Neurolucida 360, neuTube, or Imaris software. KEY RESULTS: The transduction of multicolor AAV vectors induced random sparse spectral labeling of soma and neurites primarily in the myenteric plexus of the proximal colon, while neurons in the submucosal plexus were occasionally transduced. Digital tracing in 3D showed various types of wiring, including multiple conjunctions of one neuron with other neurons, neurites en route, and endings; clusters of neurons in close apposition between each other; axon-axon parallel conjunctions; and intraganglionic nerve endings consisting of multiple nerve endings and passing fibers. Most of digitally traced neuronal somas were of small or medium in size. CONCLUSIONS & INFERENCES: The multicolor AAV-PHP.S-packaged vectors enabled random sparse spectral labeling and revealed complexities of enteric microcircuit in the mouse proximal colon. The techniques can facilitate digital modeling of enteric micro-circuitry.
Assuntos
Colo/metabolismo , Sistema Nervoso Entérico/metabolismo , Plexo Submucoso/metabolismo , Animais , Colo/inervação , Dependovirus , Sistema Nervoso Entérico/virologia , Feminino , Técnicas de Transferência de Genes , Proteínas de Fluorescência Verde , Masculino , Camundongos , Plexo Submucoso/virologiaRESUMO
BACKGROUND: The correlation between the expression levels of transcription factors and their target genes can be used to infer interactions within animal regulatory networks, but current methods are limited in their ability to make correct predictions. RESULTS: Here we describe a novel approach which uses nonparametric statistics to generate ordinary differential equation (ODE) models from expression data. Compared to other dynamical methods, our approach requires minimal information about the mathematical structure of the ODE; it does not use qualitative descriptions of interactions within the network; and it employs new statistics to protect against over-fitting. It generates spatio-temporal maps of factor activity, highlighting the times and spatial locations at which different regulators might affect target gene expression levels. We identify an ODE model for eve mRNA pattern formation in the Drosophila melanogaster blastoderm and show that this reproduces the experimental patterns well. Compared to a non-dynamic, spatial-correlation model, our ODE gives 59% better agreement to the experimentally measured pattern. Our model suggests that protein factors frequently have the potential to behave as both an activator and inhibitor for the same cis-regulatory module depending on the factors' concentration, and implies different modes of activation and repression. CONCLUSIONS: Our method provides an objective quantification of the regulatory potential of transcription factors in a network, is suitable for both low- and moderate-dimensional gene expression datasets, and includes improvements over existing dynamic and static models.
Assuntos
Drosophila melanogaster/embriologia , Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Modelos Biológicos , Animais , Blastoderma , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/genética , Proteínas/genética , Fatores de Transcrição/genética , Transcrição GênicaRESUMO
Complex spatiotemporal gene expression patterns direct the development of the fertilized egg into an adult animal. Comparisons across species show that, in spite of changes in the underlying regulatory DNA sequence, developmental programs can be maintained across millions of years of evolution. Reciprocally, changes in gene expression can be used to generate morphological novelty. Distinguishing between changes in regulatory DNA that lead to changes in gene expression and those that do not is therefore a central goal of evolutionary developmental biology. Quantitative, spatially-resolved measurements of developmental gene expression patterns play a crucial role in this goal, enabling the detection of subtle phenotypic differences between species and the development of computations models that link the sequence of regulatory DNA to expression patterns. Here we report the generation of two atlases of cellular resolution gene expression measurements for the primary anterior-posterior patterning genes in Drosophila simulans and Drosophila virilis By combining these data sets with existing atlases for three other Drosophila species, we detect subtle differences in the gene expression patterns and dynamics driving the highly conserved axis patterning system and delineate inter-species differences in the embryonic morphology. These data sets will be a resource for future modeling studies of the evolution of developmental gene regulatory networks.
Assuntos
Padronização Corporal , Drosophila/embriologia , Desenvolvimento Embrionário , Animais , Biomarcadores , Padronização Corporal/genética , Embrião não Mamífero , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Redes Reguladoras de Genes , Especificidade de Órgãos , Especificidade da Espécie , TranscriptomaRESUMO
Heart rate is under the precise control of the autonomic nervous system. However, the wiring of peripheral neural circuits that regulate heart rate is poorly understood. Here, we develop a clearing-imaging-analysis pipeline to visualize innervation of intact hearts in 3D and employed a multi-technique approach to map parasympathetic and sympathetic neural circuits that control heart rate in mice. We identify cholinergic neurons and noradrenergic neurons in an intrinsic cardiac ganglion and the stellate ganglia, respectively, that project to the sinoatrial node. We also report that the heart rate response to optogenetic versus electrical stimulation of the vagus nerve displays different temporal characteristics and that vagal afferents enhance parasympathetic and reduce sympathetic tone to the heart via central mechanisms. Our findings provide new insights into neural regulation of heart rate, and our methodology to study cardiac circuits can be readily used to interrogate neural control of other visceral organs.
Assuntos
Frequência Cardíaca/fisiologia , Neurônios Motores/fisiologia , Animais , Neurônios Colinérgicos/metabolismo , Neurônios Colinérgicos/fisiologia , Eletrofisiologia , Feminino , Masculino , Camundongos , Sistema Nervoso Periférico/metabolismo , Sistema Nervoso Periférico/fisiologia , Nervo Vago/metabolismo , Nervo Vago/fisiologiaRESUMO
Bioinspiration-using insights into the function of biological systems for the development of new engineering concepts-is already a successful and rapidly growing field. However, only a small portion of the world's biodiversity has thus far been considered as a potential source for engineering inspiration. This means that vast numbers of biological systems of potentially high value to engineering have likely gone unnoticed. Even more important, insights into form and function that reside in the evolutionary relationships across the tree of life have not yet received attention by engineers. These insights could soon become accessible through recent developments in disparate areas of research; in particular, advancements in digitization of museum specimens, methods to describe and analyze complex biological shapes, quantitative prediction of biological function from form, and analysis of large digital data sets. Taken together, these emerging capabilities should make it possible to mine the world's known biodiversity as a natural resource for knowledge relevant to engineering. This transformation of bioinspiration would be very timely in the development of engineering, because it could yield exactly the kind of insights that are needed to make technology more autonomous, adaptive, and capable of operation in complex environments.
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Engenharia/métodos , Pesquisa , Tecnologia/métodosRESUMO
Figure-ground organization refers to the visual perception that a contour separating two regions belongs to one of the regions. Recent studies have found neural correlates of figure-ground assignment in V2 as early as 10-25 ms after response onset, providing strong support for the role of local bottom-up processing. How much information about figure-ground assignment is available from locally computed cues? Using a large collection of natural images, in which neighboring regions were assigned a figure-ground relation by human observers, we quantified the extent to which figural regions locally tend to be smaller, more convex, and lie below ground regions. Our results suggest that these Gestalt cues are ecologically valid, and we quantify their relative power. We have also developed a simple bottom-up computational model of figure-ground assignment that takes image contours as input. Using parameters fit to natural image statistics, the model is capable of matching human-level performance when scene context limited.
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Sinais (Psicologia) , Área de Dependência-Independência , Natureza , Estimulação Luminosa/métodos , Percepção Visual , Humanos , Luz , Modelos PsicológicosRESUMO
Discriminating between black and white spruce (Picea mariana and Picea glauca) is a difficult palynological classification problem that, if solved, would provide valuable data for paleoclimate reconstructions. We developed an open-source visual recognition software (ARLO, Automated Recognition with Layered Optimization) capable of differentiating between these two species at an accuracy on par with human experts. The system applies pattern recognition and machine learning to the analysis of pollen images and discovers general-purpose image features, defined by simple features of lines and grids of pixels taken at different dimensions, size, spacing, and resolution. It adapts to a given problem by searching for the most effective combination of both feature representation and learning strategy. This results in a powerful and flexible framework for image classification. We worked with images acquired using an automated slide scanner. We first applied a hash-based "pollen spotting" model to segment pollen grains from the slide background. We next tested ARLO's ability to reconstruct black to white spruce pollen ratios using artificially constructed slides of known ratios. We then developed a more scalable hash-based method of image analysis that was able to distinguish between the pollen of black and white spruce with an estimated accuracy of 83.61%, comparable to human expert performance. Our results demonstrate the capability of machine learning systems to automate challenging taxonomic classifications in pollen analysis, and our success with simple image representations suggests that our approach is generalizable to many other object recognition problems.