Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 116
Filtrar
Mais filtros

Base de dados
Tipo de documento
Intervalo de ano de publicação
1.
Cell ; 183(6): 1586-1599.e10, 2020 12 10.
Artigo em Inglês | MEDLINE | ID: mdl-33159859

RESUMO

The hippocampus is crucial for spatial navigation and episodic memory formation. Hippocampal place cells exhibit spatially selective activity within an environment and have been proposed to form the neural basis of a cognitive map of space that supports these mnemonic functions. However, the direct influence of place cell activity on spatial navigation behavior has not yet been demonstrated. Using an 'all-optical' combination of simultaneous two-photon calcium imaging and two-photon optogenetics, we identified and selectively activated place cells that encoded behaviorally relevant locations in a virtual reality environment. Targeted stimulation of a small number of place cells was sufficient to bias the behavior of animals during a spatial memory task, providing causal evidence that hippocampal place cells actively support spatial navigation and memory.


Assuntos
Hipocampo/citologia , Células de Lugar/citologia , Comportamento Espacial , Memória Espacial , Animais , Comportamento Animal , Masculino , Camundongos Endogâmicos C57BL , Neurônios/metabolismo , Opsinas/metabolismo , Optogenética , Fótons , Recompensa , Corrida , Navegação Espacial
3.
Nature ; 617(7962): 769-776, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-37138089

RESUMO

Sensory processing in the neocortex requires both feedforward and feedback information flow between cortical areas1. In feedback processing, higher-level representations provide contextual information to lower levels, and facilitate perceptual functions such as contour integration and figure-ground segmentation2,3. However, we have limited understanding of the circuit and cellular mechanisms that mediate feedback influence. Here we use long-range all-optical connectivity mapping in mice to show that feedback influence from the lateromedial higher visual area (LM) to the primary visual cortex (V1) is spatially organized. When the source and target of feedback represent the same area of visual space, feedback is relatively suppressive. By contrast, when the source is offset from the target in visual space, feedback is relatively facilitating. Two-photon calcium imaging data show that this facilitating feedback is nonlinearly integrated in the apical tuft dendrites of V1 pyramidal neurons: retinotopically offset (surround) visual stimuli drive local dendritic calcium signals indicative of regenerative events, and two-photon optogenetic activation of LM neurons projecting to identified feedback-recipient spines in V1 can drive similar branch-specific local calcium signals. Our results show how neocortical feedback connectivity and nonlinear dendritic integration can together form a substrate to support both predictive and cooperative contextual interactions.


Assuntos
Dendritos , Retroalimentação Fisiológica , Córtex Visual , Vias Visuais , Animais , Camundongos , Cálcio/metabolismo , Dendritos/fisiologia , Córtex Visual/citologia , Córtex Visual/fisiologia , Vias Visuais/citologia , Vias Visuais/fisiologia , Retroalimentação Fisiológica/fisiologia , Córtex Visual Primário/citologia , Córtex Visual Primário/fisiologia , Células Piramidais/citologia , Células Piramidais/fisiologia , Optogenética , Sinalização do Cálcio
5.
Proc Natl Acad Sci U S A ; 118(30)2021 07 27.
Artigo em Inglês | MEDLINE | ID: mdl-34301882

RESUMO

The dendrites of neocortical pyramidal neurons are excitable. However, it is unknown how synaptic inputs engage nonlinear dendritic mechanisms during sensory processing in vivo, and how they in turn influence action potential output. Here, we provide a quantitative account of the relationship between synaptic inputs, nonlinear dendritic events, and action potential output. We developed a detailed pyramidal neuron model constrained by in vivo dendritic recordings. We drive this model with realistic input patterns constrained by sensory responses measured in vivo and connectivity measured in vitro. We show mechanistically that under realistic conditions, dendritic Na+ and NMDA spikes are the major determinants of neuronal output in vivo. We demonstrate that these dendritic spikes can be triggered by a surprisingly small number of strong synaptic inputs, in some cases even by single synapses. We predict that dendritic excitability allows the 1% strongest synaptic inputs of a neuron to control the tuning of its output. Active dendrites therefore allow smaller subcircuits consisting of only a few strongly connected neurons to achieve selectivity for specific sensory features.


Assuntos
Potenciais de Ação , Dendritos/fisiologia , Modelos Neurológicos , Neurônios/fisiologia , Células Piramidais/fisiologia , Sinapses/fisiologia , Transmissão Sináptica , Animais , Sinalização do Cálcio , Potenciais Pós-Sinápticos Excitadores , Camundongos , N-Metilaspartato/metabolismo , Orientação , Ratos , Sódio/metabolismo
6.
Nature ; 551(7679): 232-236, 2017 11 08.
Artigo em Inglês | MEDLINE | ID: mdl-29120427

RESUMO

Sensory, motor and cognitive operations involve the coordinated action of large neuronal populations across multiple brain regions in both superficial and deep structures. Existing extracellular probes record neural activity with excellent spatial and temporal (sub-millisecond) resolution, but from only a few dozen neurons per shank. Optical Ca2+ imaging offers more coverage but lacks the temporal resolution needed to distinguish individual spikes reliably and does not measure local field potentials. Until now, no technology compatible with use in unrestrained animals has combined high spatiotemporal resolution with large volume coverage. Here we design, fabricate and test a new silicon probe known as Neuropixels to meet this need. Each probe has 384 recording channels that can programmably address 960 complementary metal-oxide-semiconductor (CMOS) processing-compatible low-impedance TiN sites that tile a single 10-mm long, 70 × 20-µm cross-section shank. The 6 × 9-mm probe base is fabricated with the shank on a single chip. Voltage signals are filtered, amplified, multiplexed and digitized on the base, allowing the direct transmission of noise-free digital data from the probe. The combination of dense recording sites and high channel count yielded well-isolated spiking activity from hundreds of neurons per probe implanted in mice and rats. Using two probes, more than 700 well-isolated single neurons were recorded simultaneously from five brain structures in an awake mouse. The fully integrated functionality and small size of Neuropixels probes allowed large populations of neurons from several brain structures to be recorded in freely moving animals. This combination of high-performance electrode technology and scalable chip fabrication methods opens a path towards recording of brain-wide neural activity during behaviour.


Assuntos
Eletrodos , Neurônios/fisiologia , Silício/metabolismo , Animais , Córtex Entorrinal/citologia , Córtex Entorrinal/fisiologia , Feminino , Masculino , Camundongos , Movimento/fisiologia , Córtex Pré-Frontal/citologia , Córtex Pré-Frontal/fisiologia , Ratos , Semicondutores , Vigília/fisiologia
7.
PLoS Biol ; 17(9): e3000414, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31479441

RESUMO

Bardet-Biedl syndrome (BBS), a ciliopathy, is a rare genetic condition characterised by retinal degeneration, obesity, kidney failure, and cognitive impairment. In spite of progress made in our general understanding of BBS aetiology, the molecular and cellular mechanisms underlying cognitive impairment in BBS remain elusive. Here, we report that the loss of BBS proteins causes synaptic dysfunction in principal neurons, providing a possible explanation for the cognitive impairment phenotype observed in BBS patients. Using synaptosomal proteomics and immunocytochemistry, we demonstrate the presence of Bbs proteins in the postsynaptic density (PSD) of hippocampal neurons. Loss of Bbs results in a significant reduction of dendritic spines in principal neurons of Bbs mouse models. Furthermore, we show that spine deficiency correlates with events that destabilise spine architecture, such as impaired spine membrane receptor signalling, known to be involved in the maintenance of dendritic spines. Our findings suggest a role for BBS proteins in dendritic spine homeostasis that may be linked to the cognitive phenotype observed in BBS.


Assuntos
Síndrome de Bardet-Biedl/patologia , Proteínas do Citoesqueleto/metabolismo , Espinhas Dendríticas/patologia , Animais , Ansiedade , Síndrome de Bardet-Biedl/metabolismo , Síndrome de Bardet-Biedl/fisiopatologia , Síndrome de Bardet-Biedl/psicologia , Giro Denteado/fisiopatologia , Modelos Animais de Doenças , Potenciais Pós-Sinápticos Excitadores , Feminino , Masculino , Memória , Camundongos , Receptor IGF Tipo 1/metabolismo , Sinaptossomos/metabolismo
9.
Nat Methods ; 15(12): 1037-1040, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30420686

RESUMO

Understanding the causal relationship between neural activity and behavior requires the ability to perform rapid and targeted interventions in ongoing activity. Here we describe a closed-loop all-optical strategy for dynamically controlling neuronal activity patterns in awake mice. We rapidly tailored and delivered two-photon optogenetic stimulation based on online readout of activity using simultaneous two-photon imaging, thus enabling the manipulation of neural circuit activity 'on the fly' during behavior.


Assuntos
Rede Nervosa/fisiologia , Neurônios/fisiologia , Imagem Óptica/métodos , Optogenética/instrumentação , Optogenética/métodos , Animais , Estimulação Elétrica , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Processamento de Sinais Assistido por Computador
11.
Nature ; 493(7430): 97-100, 2013 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-23172139

RESUMO

The activity of the cerebral cortex is thought to depend on the precise relationship between synaptic excitation and inhibition. In the visual cortex, in particular, intracellular measurements have related response selectivity to coordinated increases in excitation and inhibition. These measurements, however, have all been made during anaesthesia, which strongly influences cortical state and therefore sensory processing. The synaptic activity that is evoked by visual stimulation during wakefulness is unknown. Here we measured visually evoked responses--and the underlying synaptic conductances--in the visual cortex of anaesthetized and awake mice. Under anaesthesia, responses could be elicited from a large region of visual space and were prolonged. During wakefulness, responses were more spatially selective and much briefer. Whole-cell patch-clamp recordings of synaptic conductances showed a difference in synaptic inhibition between the two conditions. Under anaesthesia, inhibition tracked excitation in amplitude and spatial selectivity. By contrast, during wakefulness, inhibition was much stronger than excitation and had extremely broad spatial selectivity. We conclude that during wakefulness, cortical responses to visual stimulation are dominated by synaptic inhibition, restricting the spatial spread and temporal persistence of neural activity. These results provide a direct glimpse of synaptic mechanisms that control sensory responses in the awake cortex.


Assuntos
Inibição Neural/fisiologia , Córtex Visual/fisiologia , Vigília/fisiologia , Anestesia , Animais , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Modelos Neurológicos , Técnicas de Patch-Clamp , Estimulação Luminosa , Sinapses/metabolismo , Transmissão Sináptica , Fatores de Tempo
12.
Nature ; 503(7474): 115-20, 2013 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-24162850

RESUMO

Neuronal dendrites are electrically excitable: they can generate regenerative events such as dendritic spikes in response to sufficiently strong synaptic input. Although such events have been observed in many neuronal types, it is not well understood how active dendrites contribute to the tuning of neuronal output in vivo. Here we show that dendritic spikes increase the selectivity of neuronal responses to the orientation of a visual stimulus (orientation tuning). We performed direct patch-clamp recordings from the dendrites of pyramidal neurons in the primary visual cortex of lightly anaesthetized and awake mice, during sensory processing. Visual stimulation triggered regenerative local dendritic spikes that were distinct from back-propagating action potentials. These events were orientation tuned and were suppressed by either hyperpolarization of membrane potential or intracellular blockade of NMDA (N-methyl-d-aspartate) receptors. Both of these manipulations also decreased the selectivity of subthreshold orientation tuning measured at the soma, thus linking dendritic regenerative events to somatic orientation tuning. Together, our results suggest that dendritic spikes that are triggered by visual input contribute to a fundamental cortical computation: enhancing orientation selectivity in the visual cortex. Thus, dendritic excitability is an essential component of behaviourally relevant computations in neurons.


Assuntos
Potenciais de Ação , Dendritos/fisiologia , Córtex Visual/citologia , Animais , Sinalização do Cálcio , Sedação Consciente , Potenciais Evocados/fisiologia , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Técnicas de Patch-Clamp , Estimulação Luminosa , Células Piramidais/citologia , Células Piramidais/fisiologia , Receptores de N-Metil-D-Aspartato/metabolismo , Vigília/fisiologia
13.
Nat Methods ; 12(2): 140-6, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25532138

RESUMO

We describe an all-optical strategy for simultaneously manipulating and recording the activity of multiple neurons with cellular resolution in vivo. We performed simultaneous two-photon optogenetic activation and calcium imaging by coexpression of a red-shifted opsin and a genetically encoded calcium indicator. A spatial light modulator allows tens of user-selected neurons to be targeted for spatiotemporally precise concurrent optogenetic activation, while simultaneous fast calcium imaging provides high-resolution network-wide readout of the manipulation with negligible optical cross-talk. Proof-of-principle experiments in mouse barrel cortex demonstrate interrogation of the same neuronal population during different behavioral states and targeting of neuronal ensembles based on their functional signature. This approach extends the optogenetic toolkit beyond the specificity obtained with genetic or viral approaches, enabling high-throughput, flexible and long-term optical interrogation of functionally defined neural circuits with single-cell and single-spike resolution in the mouse brain in vivo.


Assuntos
Encéfalo/fisiologia , Sinalização do Cálcio/fisiologia , Neurônios/fisiologia , Optogenética , Potenciais de Ação/genética , Potenciais de Ação/fisiologia , Animais , Comportamento Animal/fisiologia , Encéfalo/metabolismo , Cálcio/metabolismo , Sinalização do Cálcio/genética , Proteínas de Ligação ao Cálcio/genética , Feminino , Locomoção/genética , Locomoção/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microscopia de Fluorescência por Excitação Multifotônica , Neurônios/metabolismo , Opsinas/genética , Estimulação Luminosa , Análise de Célula Única
14.
Proc Natl Acad Sci U S A ; 112(42): 13099-104, 2015 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-26432880

RESUMO

Classical feed-forward inhibition involves an excitation-inhibition sequence that enhances the temporal precision of neuronal responses by narrowing the window for synaptic integration. In the input layer of the cerebellum, feed-forward inhibition is thought to preserve the temporal fidelity of granule cell spikes during mossy fiber stimulation. Although this classical feed-forward inhibitory circuit has been demonstrated in vitro, the extent to which inhibition shapes granule cell sensory responses in vivo remains unresolved. Here we combined whole-cell patch-clamp recordings in vivo and dynamic clamp recordings in vitro to directly assess the impact of Golgi cell inhibition on sensory information transmission in the granule cell layer of the cerebellum. We show that the majority of granule cells in Crus II of the cerebrocerebellum receive sensory-evoked phasic and spillover inhibition prior to mossy fiber excitation. This preceding inhibition reduces granule cell excitability and sensory-evoked spike precision, but enhances sensory response reproducibility across the granule cell population. Our findings suggest that neighboring granule cells and Golgi cells can receive segregated and functionally distinct mossy fiber inputs, enabling Golgi cells to regulate the size and reproducibility of sensory responses.


Assuntos
Cerebelo/fisiologia , Grânulos Citoplasmáticos/fisiologia , Complexo de Golgi/fisiologia , Animais , Técnicas de Patch-Clamp , Ratos , Ratos Sprague-Dawley
15.
PLoS Comput Biol ; 12(8): e1005000, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27541958

RESUMO

Purkinje neurons play an important role in cerebellar computation since their axons are the only projection from the cerebellar cortex to deeper cerebellar structures. They have complex internal dynamics, which allow them to fire spontaneously, display bistability, and also to be involved in network phenomena such as high frequency oscillations and travelling waves. Purkinje cells exhibit type II excitability, which can be revealed by a discontinuity in their f-I curves. We show that this excitability mechanism allows Purkinje cells to be efficiently inhibited by noise of a particular variance, a phenomenon known as inverse stochastic resonance (ISR). While ISR has been described in theoretical models of single neurons, here we provide the first experimental evidence for this effect. We find that an adaptive exponential integrate-and-fire model fitted to the basic Purkinje cell characteristics using a modified dynamic IV method displays ISR and bistability between the resting state and a repetitive activity limit cycle. ISR allows the Purkinje cell to operate in different functional regimes: the all-or-none toggle or the linear filter mode, depending on the variance of the synaptic input. We propose that synaptic noise allows Purkinje cells to quickly switch between these functional regimes. Using mutual information analysis, we demonstrate that ISR can lead to a locally optimal information transfer between the input and output spike train of the Purkinje cell. These results provide the first experimental evidence for ISR and suggest a functional role for ISR in cerebellar information processing.


Assuntos
Potenciais de Ação/fisiologia , Modelos Neurológicos , Células de Purkinje/citologia , Células de Purkinje/fisiologia , Animais , Biologia Computacional , Ratos , Ratos Sprague-Dawley , Processos Estocásticos
16.
Nature ; 539(7628): 159-161, 2016 11 10.
Artigo em Inglês | MEDLINE | ID: mdl-27830818
17.
J Neurosci ; 35(41): 13917-26, 2015 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-26468193

RESUMO

There have been two recent revolutionary advances in neuroscience: First, genetically encoded activity sensors have brought the goal of optical detection of single action potentials in vivo within reach. Second, optogenetic actuators now allow the activity of neurons to be controlled with millisecond precision. These revolutions have now been combined, together with advanced microscopies, to allow "all-optical" readout and manipulation of activity in neural circuits with single-spike and single-neuron precision. This is a transformational advance that will open new frontiers in neuroscience research. Harnessing the power of light in the all-optical approach requires coexpression of genetically encoded activity sensors and optogenetic probes in the same neurons, as well as the ability to simultaneously target and record the light from the selected neurons. It has recently become possible to combine sensors and optical strategies that are sufficiently sensitive and cross talk free to enable single-action-potential sensitivity and precision for both readout and manipulation in the intact brain. The combination of simultaneous readout and manipulation from the same genetically defined cells will enable a wide range of new experiments as well as inspire new technologies for interacting with the brain. The advances described in this review herald a future where the traditional tools used for generations by physiologists to study and interact with the brain-stimulation and recording electrodes-can largely be replaced by light. We outline potential future developments in this field and discuss how the all-optical strategy can be applied to solve fundamental problems in neuroscience. SIGNIFICANCE STATEMENT: This review describes the nexus of dramatic recent developments in optogenetic probes, genetically encoded activity sensors, and novel microscopies, which together allow the activity of neural circuits to be recorded and manipulated entirely using light. The optical and protein engineering strategies that form the basis of this "all-optical" approach are now sufficiently advanced to enable single-neuron and single-action potential precision for simultaneous readout and manipulation from the same functionally defined neurons in the intact brain. These advances promise to illuminate many fundamental challenges in neuroscience, including transforming our search for the neural code and the links between neural circuit activity and behavior.


Assuntos
Encéfalo/citologia , Rede Nervosa/fisiologia , Neurônios/fisiologia , Optogenética , Animais , Humanos
18.
Nature ; 466(7302): 123-7, 2010 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-20596024

RESUMO

It is well known that neural activity exhibits variability, in the sense that identical sensory stimuli produce different responses, but it has been difficult to determine what this variability means. Is it noise, or does it carry important information-about, for example, the internal state of the organism? Here we address this issue from the bottom up, by asking whether small perturbations to activity in cortical networks are amplified. Based on in vivo whole-cell patch-clamp recordings in rat barrel cortex, we find that a perturbation consisting of a single extra spike in one neuron produces approximately 28 additional spikes in its postsynaptic targets. We also show, using simultaneous intra- and extracellular recordings, that a single spike in a neuron produces a detectable increase in firing rate in the local network. Theoretical analysis indicates that this amplification leads to intrinsic, stimulus-independent variations in membrane potential of the order of +/-2.2-4.5 mV-variations that are pure noise, and so carry no information at all. Therefore, for the brain to perform reliable computations, it must either use a rate code, or generate very large, fast depolarizing events, such as those proposed by the theory of synfire chains. However, in our in vivo recordings, we found that such events were very rare. Our findings are thus consistent with the idea that cortex is likely to use primarily a rate code.


Assuntos
Córtex Cerebral/fisiologia , Modelos Neurológicos , Potenciais de Ação/fisiologia , Animais , Artefatos , Córtex Cerebral/citologia , Neurônios/metabolismo , Técnicas de Patch-Clamp , Probabilidade , Ratos , Ratos Sprague-Dawley , Processos Estocásticos
19.
Nature ; 461(7266): 930-9, 2009 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-19829373

RESUMO

Electrophysiology, the 'gold standard' for investigating neuronal signalling, is being challenged by a new generation of optical probes. Together with new forms of microscopy, these probes allow us to measure and control neuronal signals with spatial resolution and genetic specificity that already greatly surpass those of electrophysiology. We predict that the photon will progressively replace the electron for probing neuronal function, particularly for targeted stimulation and silencing of neuronal populations. Although electrophysiological characterization of channels, cells and neural circuits will remain necessary, new combinations of electrophysiology and imaging should lead to transformational discoveries in neuroscience.


Assuntos
Eletrofisiologia/métodos , Luz , Neurociências/métodos , Óptica e Fotônica/métodos , Animais , Cálcio/metabolismo , Eletrofisiologia/instrumentação , Eletrofisiologia/tendências
20.
Proc Natl Acad Sci U S A ; 109(27): 11014-8, 2012 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-22715290

RESUMO

The wide diversity of dendritic trees is one of the most striking features of neural circuits. Here we develop a general quantitative theory relating the total length of dendritic wiring to the number of branch points and synapses. We show that optimal wiring predicts a 2/3 power law between these measures. We demonstrate that the theory is consistent with data from a wide variety of neurons across many different species and helps define the computational compartments in dendritic trees. Our results imply fundamentally distinct design principles for dendritic arbors compared with vascular, bronchial, and botanical trees.


Assuntos
Dendritos/fisiologia , Modelos Neurológicos , Vias Neurais/fisiologia , Neurônios/fisiologia , Sinapses/fisiologia , Animais , Humanos , Neurônios/ultraestrutura
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA