RESUMO
The optomotor response (OMR) is central to the locomotory behavior in diverse animal species including insects, fish and mammals. Furthermore, the study of the OMR in larval zebrafish has become a key model system for investigating the neural basis of sensorimotor control. However, a comprehensive understanding of the underlying control algorithms is still outstanding. In fish it is often assumed that the OMR, by reducing average optic flow across the retina, serves to stabilize position with respect to the ground. Yet the degree to which this is achieved, and how it could emerge from the intermittent burst dynamics of larval zebrafish swimming, are unclear. Here, we combine detailed computational modeling with a new approach to free-swimming experiments in which we control the amount of visual feedback produced by a given motor effort by varying the height of the larva above a moving grid stimulus. We develop an account of underlying feedback control mechanisms that describes both the bout initiation process and the control of swim speed during bouts. We observe that the degree to which fish stabilize their position is only partial and height-dependent, raising questions about its function. We find the relative speed profile during bouts follows a fixed temporal pattern independent of absolute bout speed, suggesting that bout speed and bout termination are not separately controlled. We also find that the reverse optic flow, experienced when the fish is swimming faster than the stimulus, plays a minimal role in control of the OMR despite carrying most of the sensory information about self-movement. These results shed new light on the underlying dynamics of the OMR in larval zebrafish and will be crucial for future work aimed at identifying the neural basis of this behavior.
Assuntos
Natação , Peixe-Zebra , Animais , Peixe-Zebra/fisiologia , Larva/fisiologia , Natação/fisiologia , Atividade Motora/fisiologia , Algoritmos , MamíferosRESUMO
The vertebrate inner retina is driven by photoreceptors whose outputs are already pre-processed; in zebrafish, outer retinal circuits split "color" from "grayscale" information across four cone-photoreceptor types. It remains unclear how the inner retina processes incoming spectral information while also combining cone signals to shape grayscale functions. We address this question by imaging the light-driven responses of amacrine cells (ACs) and bipolar cells (BCs) in larval zebrafish in the presence and pharmacological absence of inner retinal inhibition. We find that ACs enhance opponency in some bipolar cells while at the same time suppressing pre-existing opponency in others, so that, depending on the retinal region, the net change in the number of color-opponent units is essentially zero. To achieve this "dynamic balance," ACs counteract intrinsic color opponency of BCs via the On channel. Consistent with these observations, Off-stratifying ACs are exclusively achromatic, while all color-opponent ACs stratify in the On sublamina.
Assuntos
Células Amácrinas , Peixe-Zebra , Animais , Células Amácrinas/fisiologia , Retina/fisiologia , Células Fotorreceptoras Retinianas Cones/fisiologiaRESUMO
The statistics of vesicle release determine how synapses transfer information, but the classical Poisson model of independent release does not always hold at the first stages of vision and hearing. There, ribbon synapses also encode sensory signals as events comprising two or more vesicles released simultaneously. The implications of such coordinated multivesicular release (MVR) for spike generation are not known. Here we investigate how MVR alters the transmission of sensory information compared with Poisson synapses using a pure rate-code. We used leaky integrate-and-fire models incorporating the statistics of release measured experimentally from glutamatergic synapses of retinal bipolar cells in zebrafish (both sexes) and compared these with models assuming Poisson inputs constrained to operate at the same average rates. We find that MVR can increase the number of spikes generated per vesicle while reducing interspike intervals and latency to first spike. The combined effect was to increase the efficiency of information transfer (bits per vesicle) over a range of conditions mimicking target neurons of different size. MVR was most advantageous in neurons with short time constants and reliable synaptic inputs, when less convergence was required to trigger spikes. In the special case of a single input driving a neuron, as occurs in the auditory system of mammals, MVR increased information transfer whenever spike generation required more than one vesicle. This study demonstrates how presynaptic integration of vesicles by MVR can increase the efficiency with which sensory information is transmitted compared with a rate-code described by Poisson statistics.SIGNIFICANCE STATEMENT Neurons communicate by the stochastic release of vesicles at the synapse and the statistics of this process will determine how information is represented by spikes. The classical model is that vesicles are released independently by a Poisson process, but this does not hold at ribbon-type synapses specialized to transmit the first electrical signals in vision and hearing, where two or more vesicles can fuse in a single event by a process termed coordinated multivesicular release. This study shows that multivesicular release can increase the number of spikes generated per vesicle and the efficiency of information transfer (bits per vesicle) over a range of conditions found in the retina and peripheral auditory system.
Assuntos
Vesículas Sinápticas , Peixe-Zebra , Masculino , Animais , Feminino , Vesículas Sinápticas/fisiologia , Sinapses/fisiologia , Células Bipolares da Retina , Retina/fisiologia , Transmissão Sináptica/fisiologia , MamíferosRESUMO
Neuromodulators adapt sensory circuits to changes in the external world or the animal's internal state and synapses are key control sites for such plasticity. Less clear is how neuromodulation alters the amount of information transmitted through the circuit. We investigated this question in the context of the diurnal regulation of visual processing in the retina of zebrafish, focusing on ribbon synapses of bipolar cells. We demonstrate that contrast-sensitivity peaks in the afternoon accompanied by a four-fold increase in the average Shannon information transmitted from an active zone. This increase reflects higher synaptic gain, lower spontaneous "noise" and reduced variability of evoked responses. Simultaneously, an increase in the probability of multivesicular events with larger information content increases the efficiency of transmission (bits per vesicle) by factors of 1.5-2.7. This study demonstrates the multiplicity of mechanisms by which a neuromodulator can adjust the synaptic transfer of sensory information.
Assuntos
Transmissão Sináptica , Peixe-Zebra , Animais , Neurotransmissores , Retina/fisiologia , Sinapses/fisiologia , Transmissão Sináptica/fisiologiaRESUMO
Sensory processing in the cortex adapts to the history of stimulation but the mechanisms are not understood. Imaging the primary visual cortex of mice we find here that an increase in stimulus contrast is not followed by a simple decrease in gain of pyramidal cells; as many cells increase gain to improve detection of a subsequent decrease in contrast. Depressing and sensitizing forms of adaptation also occur in different types of interneurons (PV, SST and VIP) and the net effect within individual pyramidal cells reflects the balance of PV inputs, driving depression, and a subset of SST interneurons driving sensitization. Changes in internal state associated with locomotion increase gain across the population of pyramidal cells while maintaining the balance between these opposite forms of plasticity, consistent with activation of both VIP->SST and SST->PV disinhibitory pathways. These results reveal how different inhibitory microcircuits adjust the gain of pyramidal cells signalling changes in stimulus strength.
Assuntos
Córtex Visual , Animais , Córtex Cerebral/metabolismo , Interneurônios/fisiologia , Locomoção , Camundongos , Parvalbuminas/metabolismo , Células Piramidais/fisiologia , Córtex Visual/fisiologiaRESUMO
BACKGROUND: Selective Plane Illumination Microscopy (SPIM) is a fluorescence imaging technique that allows volumetric imaging at high spatio-temporal resolution to monitor neural activity in live organisms such as larval zebrafish. A major challenge in the construction of a custom SPIM microscope using a scanned laser beam is the control and synchronization of the various hardware components. NEW METHOD: We present an open-source software, µSPIM Toolset, built around the widely adopted MicroManager platform, that provides control and acquisition functionality for a SPIM. A key advantage of µSPIM Toolset is a series of calibration procedures that optimize acquisition for a given set-up, making it relatively independent of the optical design of the microscope or the hardware used to build it. RESULTS: µSPIM Toolset allows imaging of calcium activity throughout the brain of larval zebrafish at rates of 100 planes per second with single cell resolution. COMPARISON WITH EXISTING METHODS: Several designs of SPIM have been published but are focused on imaging of developmental processes using a slower setup with a moving stage and therefore have limited use for functional imaging. In comparison, µSPIM Toolset uses a scanned beam to allow imaging at higher acquisition frequencies while minimizing disturbance of the sample. CONCLUSIONS: The µSPIM Toolset provides a flexible solution for the control of SPIM microscopes and demonstrated its utility for brain-wide imaging of neural activity in larval zebrafish.
Assuntos
Microscopia , Peixe-Zebra , Animais , Iluminação , Imagem Óptica , SoftwareRESUMO
At various stages of the visual system, visual responses are modulated by arousal. Here, we find that in mice this modulation operates as early as in the first synapse from the retina and even in retinal axons. To measure retinal activity in the awake, intact brain, we imaged the synaptic boutons of retinal axons in the superior colliculus. Their activity depended not only on vision but also on running speed and pupil size, regardless of retinal illumination. Arousal typically reduced their visual responses and selectivity for direction and orientation. Recordings from retinal axons in the optic tract revealed that arousal modulates the firing of some retinal ganglion cells. Arousal had similar effects postsynaptically in colliculus neurons, independent of activity in the other main source of visual inputs to the colliculus, the primary visual cortex. These results indicate that arousal modulates activity at every stage of the mouse visual system.
Assuntos
Nível de Alerta/fisiologia , Axônios/fisiologia , Neurônios/fisiologia , Orientação Espacial/fisiologia , Células Ganglionares da Retina/fisiologia , Colículos Superiores/fisiologia , Animais , Axônios/metabolismo , Locomoção , Camundongos , Neurônios/citologia , Neurônios/metabolismo , Trato Óptico , Terminações Pré-Sinápticas/metabolismo , Células Ganglionares da Retina/citologia , Células Ganglionares da Retina/metabolismo , Colículos Superiores/diagnóstico por imagem , Colículos Superiores/metabolismo , Vias Visuais/fisiologia , VigíliaRESUMO
Animals must quickly adapt food-seeking strategies to locate nutrient sources in dynamically changing environments. Learned associations between food and environmental cues that predict its availability promote food-seeking behaviors. However, when such cues cease to predict food availability, animals undergo "extinction" learning, resulting in the inhibition of food-seeking responses. Repeatedly activated sets of neurons, or "neuronal ensembles," in the dorsal medial prefrontal cortex (dmPFC) are recruited following appetitive conditioning and undergo physiological adaptations thought to encode cue-reward associations. However, little is known about how the recruitment and intrinsic excitability of such dmPFC ensembles are modulated by extinction learning. Here, we used in vivo 2-Photon imaging in male Fos-GFP mice that express green fluorescent protein (GFP) in recently behaviorally activated neurons to determine the recruitment of activated pyramidal and GABAergic interneuron dmPFC ensembles during extinction. During extinction, we revealed a persistent activation of a subset of interneurons which emerged from a wider population of interneurons activated during the initial extinction session. This activation pattern was not observed in pyramidal cells, and extinction learning did not modulate the excitability properties of activated pyramidal cells. Moreover, extinction learning reduced the likelihood of reactivation of pyramidal cells activated during the initial extinction session. Our findings illuminate novel neuronal activation patterns in the dmPFC underlying extinction of food-seeking, and in particular, highlight an important role for interneuron ensembles in this inhibitory form of learning.
Assuntos
Sinais (Psicologia) , Córtex Pré-Frontal , Animais , Condicionamento Operante , Extinção Psicológica , Interneurônios , Masculino , Camundongos , Neurônios , RecompensaRESUMO
KEY POINTS: Motion artefacts associated with motor behaviour are an inevitable problem of multiphoton imaging in awake behaving animals, particularly when imaging synapses. Correction of axial motion artefacts usually requires volumetric imaging resulting in slower rates of acquisition. We describe a method to correct z-motion artefacts that is easy to implement and allows population imaging of synaptic activity while scanning a single plane in a standard multiphoton microscope. The method uses a reference volume acquired in two colour channels - an activity reporter and an anatomical marker of blood vessels. The procedure estimates the z-displacement in every frame and applies an intensity correction in which the z intensity profile for each synapse is modelled as a Moffat function. We demonstrate that the method allows synaptic calcium signals to be collected from populations of synaptic boutons in mouse primary visual cortex during locomotion. ABSTRACT: Functional imaging of head-fixed, behaving mice using two-photon imaging of fluorescent activity reporters has become a powerful tool for studying the function of the brain. Motion artefacts are an inevitable problem during such experiments and are routinely corrected for in x and y dimensions. However, axial (z) shifts of several microns can also occur, leading to intensity fluctuations in structures such as synapses that are small compared to the axial point-spread function of the microscope. Here we present a simple strategy to correct z-motion artefacts arising over the course of a time-series experiment in a single optical plane. Displacement in z was calculated using dye-filled blood vessels as an anatomical marker, providing high contrast images and accuracy to within â¼0.1 µm. The axial profiles of ROIs corresponding to synapses were described using a Moffat function and this 'ROI-spread function' used to correct activity traces on an ROI-by-ROI basis. We demonstrate the accuracy and utility of the procedures in simulation experiments using fluorescent beads and then apply them to correcting measurements of synaptic activity in populations of vasoactive-intestinal peptide (VIP) interneurons expressing the synaptic reporter SyGCaMP6f. Correction of z-motion artefacts had a substantial impact on the apparent correlation between synaptic activity and running speed, demonstrating the importance of correcting these when performing imaging experiments in awake mice.
Assuntos
Artefatos , Diagnóstico por Imagem , Animais , Encéfalo , Cabeça , Camundongos , Movimento (Física)RESUMO
How do sensory systems disambiguate events in the external world from signals generated by the animal's own motor actions? One strategy is to use an "efference copy" of the motor command to inhibit the sensory input caused by active behavior [1]. But does inhibition of self-generated inputs also block transmission of external stimuli? We investigated this question in the lateral line, a sensory system that allows fish and amphibians to detect water currents and that contributes to behaviors such as rheotaxis [2] and predator avoidance [3, 4]. This mechanical sense begins in hair cells grouped into neuromasts dotted along the animal's body [5]. Each neuromast contains two populations of hair cells, activated by deflection in either the anterior or posterior direction [6], as well as efferent fibers that are active during motor behavior to suppress afferents projecting to the brain [7-12]. To test how far the efference copy signal modulates responses to external stimuli, we imaged neural and synaptic activity in larval zebrafish during fictive swimming. We find that efferents transmit a precise copy of the motor signal and a single spike in the motor nerve can be associated with â¼50% inhibition of glutamate release. The efference copy signal acted with high selectivity on hair cells polarized to be activated by posterior deflections, as would occur during forward motion. During swimming, therefore, "push-pull" encoding of stimulus direction by afferents of opposite polarity is disrupted while still allowing a subset of hair cells to detect stimuli originating in the external world.
Assuntos
Sistema da Linha Lateral/fisiologia , Mecanorreceptores/fisiologia , Natação/fisiologia , Peixe-Zebra/fisiologia , AnimaisRESUMO
Animals selectively respond to environmental cues associated with food reward to optimize nutrient intake. Such appetitive conditioned stimulus-unconditioned stimulus (CS-US) associations are thought to be encoded in select, stable neuronal populations or neuronal ensembles, which undergo physiological modifications during appetitive conditioning. These ensembles in the medial prefrontal cortex (mPFC) control well-established, cue-evoked food seeking, but the mechanisms involved in the genesis of these ensembles are unclear. Here, we used male Fos-GFP mice that express green fluorescent protein (GFP) in recently behaviorally activated neurons, to reveal how dorsal mPFC neurons are recruited and modified to encode CS-US memory representations using an appetitive conditioning task. In the initial conditioning session, animals did not exhibit discriminated, cue-selective food seeking, but did so in later sessions indicating that a CS-US association was established. Using microprism-based in vivo 2-Photon imaging, we revealed that only a minority of neurons activated during the initial session was consistently activated throughout subsequent conditioning sessions and during cue-evoked memory recall. Notably, using ex vivo electrophysiology, we found that neurons activated following the initial session exhibited transient hyperexcitability. Chemogenetically enhancing the excitability of these neurons throughout subsequent conditioning sessions interfered with the development of reliable cue-selective food seeking, indicated by persistent, nondiscriminated performance. We demonstrate how appetitive learning consistently activates a subset of neurons to form a stable neuronal ensemble during the formation of a CS-US association. This ensemble may arise from a pool of hyperexcitable neurons activated during the initial conditioning session.SIGNIFICANCE STATEMENT Appetitive conditioning endows cues associated with food with the ability to guide food-seeking, through the formation of a food-cue association. Neuronal ensembles in the mPFC control established cue-evoked food-seeking. However, how neurons undergo physiological modifications and become part of an ensemble during conditioning remain unclear. We found that only a minority of dorsal mPFC neurons activated on the initial conditioning session became consistently activated during conditioning and memory recall. These initially activated neurons were also transiently hyperexcitable. We demonstrate the following: (1) how stable neuronal ensemble formation in the dorsal mPFC underlies appetitive conditioning; and (2) how this ensemble may arise from hyperexcitable neurons activated before the establishment of cue-evoked food seeking.
Assuntos
Comportamento Apetitivo/fisiologia , Rememoração Mental/fisiologia , Neurônios/fisiologia , Córtex Pré-Frontal/fisiologia , Animais , Condicionamento Clássico , Sinais (Psicologia) , Masculino , Camundongos , Camundongos Transgênicos , Plasticidade Neuronal/fisiologiaRESUMO
Most neurons transmit information digitally using spikes that trigger the release of synaptic vesicles with low probability. The first stages of vision and hearing are distinct in that they operate with analog signals, but it is unclear how these are recoded for synaptic transmission. By imaging the release of glutamate in live zebrafish, we demonstrate that ribbon synapses of retinal bipolar cells encode contrast through changes in both the frequency and amplitude of release events. Higher contrasts caused multiple vesicles to be released within an event, and such coding by amplitude often continued after the rate code had reached a maximum frequency. Glutamate packets equivalent to five vesicles transmitted four times as many bits of information per vesicle compared with those released individually. By discretizing analog signals into sequences of numbers up to about 11, ribbon synapses can increase the dynamic range, temporal precision and efficiency with which visual information is transmitted.
Assuntos
Células Bipolares da Retina/fisiologia , Transmissão Sináptica/fisiologia , Vesículas Sinápticas/fisiologia , Vias Visuais/fisiologia , Potenciais de Ação , Animais , Genes Reporter , Ácido Glutâmico/fisiologia , Fusão de Membrana , Técnicas de Patch-Clamp , Detecção de Sinal Psicológico , Peixe-Zebra/fisiologiaRESUMO
Ribbon synapses transmit information in sensory systems, but their development is not well understood. To test the hypothesis that ribbon assembly stabilizes nascent synapses, we performed simultaneous time-lapse imaging of fluorescently-tagged ribbons in retinal cone bipolar cells (BCs) and postsynaptic densities (PSD95-FP) of retinal ganglion cells (RGCs). Ribbons and PSD95-FP clusters were more stable when these components colocalized at synapses. However, synapse density on ON-alpha RGCs was unchanged in mice lacking ribbons (ribeye knockout). Wildtype BCs make both ribbon-containing and ribbon-free synapses with these GCs even at maturity. Ribbon assembly and cone BC-RGC synapse maintenance are thus regulated independently. Despite the absence of synaptic ribbons, RGCs continued to respond robustly to light stimuli, although quantitative examination of the responses revealed reduced frequency and contrast sensitivity.
Assuntos
Células Fotorreceptoras Retinianas Cones/fisiologia , Sinapses/metabolismo , Transmissão Sináptica/fisiologia , Animais , Células Cultivadas , Proteína 4 Homóloga a Disks-Large/genética , Proteína 4 Homóloga a Disks-Large/metabolismo , Microscopia Intravital/métodos , Luz , Proteínas Luminescentes/química , Proteínas Luminescentes/genética , Camundongos , Camundongos Transgênicos , Microscopia de Fluorescência/métodos , Estimulação Luminosa , Cultura Primária de Células , Células Bipolares da Retina/fisiologia , Células Ganglionares da Retina/fisiologia , Imagem com Lapso de Tempo/métodos , Proteína Vermelha FluorescenteRESUMO
Sensory systems must reduce the transmission of redundant information to function efficiently. One strategy is to continuously adjust the sensitivity of neurons to suppress responses to common features of the input while enhancing responses to new ones. Here we image the excitatory synaptic inputs and outputs of retinal ganglion cells to understand how such dynamic predictive coding is implemented in the analysis of spatial patterns. Synapses of bipolar cells become tuned to orientation through presynaptic inhibition, generating lateral antagonism in the orientation domain. Individual ganglion cells receive excitatory synapses tuned to different orientations, but feedforward inhibition generates a high-pass filter that only transmits the initial activation of these inputs, removing redundancy. These results demonstrate how a dynamic predictive code can be implemented by circuit motifs common to many parts of the brain.
Assuntos
Inibição Neural/fisiologia , Células Bipolares da Retina/fisiologia , Células Ganglionares da Retina/fisiologia , Percepção Espacial/fisiologia , Sinapses/fisiologia , Visão Ocular/fisiologia , Animais , Proteínas de Escherichia coli , Ácido Glutâmico/metabolismo , Proteínas de Fluorescência Verde , Larva , Imagem Óptica , Orientação Espacial , Proteínas Recombinantes de Fusão , Retina , Células Bipolares da Retina/metabolismo , Células Ganglionares da Retina/metabolismo , Sinapses/metabolismo , Vias Visuais , Peixe-ZebraRESUMO
Hair cells transmit mechanical information by converting deflection of the hair bundle into synaptic release of glutamate. We have investigated this process in the lateral line of larval zebrafish (male and female) to understand how stimuli are encoded within a neuromast. Using multiphoton microscopy in vivo, we imaged synaptic release of glutamate using the reporter iGluSnFR as well as deflections of the cupula. We found that the neuromast is composed of a functionally diverse population of hair cells. Half the hair cells signaled cupula motion in both directions from rest, either by increasing glutamate release in response to a deflection in the positive direction or by reducing release in the negative direction. The relationship between cupula deflection and glutamate release demonstrated maximum sensitivity at displacements of just â¼40 nm in the positive direction. The remaining hair cells only signaled motion in one direction and were less sensitive, extending the operating range of the neuromast beyond 1 µm. Adaptation of the synaptic output was also heterogeneous, with some hair cells generating sustained glutamate release in response to a steady deflection of the cupula and others generating transient outputs. Finally, a distinct signal encoded a return of the cupula to rest: a large and transient burst of glutamate release from hair cells unresponsive to the initial stimulus. A population of hair cells with these different sensitivities, operating ranges, and adaptive properties will allow the neuromast to encode weak stimuli while maintaining the dynamic range to signal the amplitude and duration of stronger deflections.SIGNIFICANCE STATEMENT Hair cells transmit information about mechanical stimuli by converting very small deflections of their hair bundle into changes in the release of the neurotransmitter glutamate. We have measured this input/output relation in the live fish using a fluorescent protein and find that different hair cells vary in their mechanical sensitivity and the time course of their response. These variations will allow the fish to sense the timing and duration of both very weak stimuli (â¼40 nm deflections) and strong stimuli (â¼1 µm), underlying the ability of the fish to avoid predators and maintain its body position in flowing water.
Assuntos
Sistema da Linha Lateral/fisiologia , Mecanorreceptores/fisiologia , Estimulação Física , Células Receptoras Sensoriais/fisiologia , Peixe-Zebra/fisiologia , Animais , Feminino , Ácido Glutâmico/fisiologia , Processamento de Imagem Assistida por Computador , Larva , Sistema da Linha Lateral/citologia , Masculino , Percepção de Movimento/fisiologia , Neuroimagem , Sinapses/fisiologiaRESUMO
Sensory processing can be tuned by a neuron's integration area, the types of inputs, and the proportion and number of connections with those inputs. Integration areas often vary topographically to sample space differentially across regions. Here, we highlight two visual circuits in which topographic changes in the postsynaptic retinal ganglion cell (RGC) dendritic territories and their presynaptic bipolar cell (BC) axonal territories are either matched or unmatched. Despite this difference, in both circuits, the proportion of inputs from each BC type, i.e., synaptic convergence between specific BCs and RGCs, remained constant across varying dendritic territory sizes. Furthermore, synapse density between BCs and RGCs was invariant across topography. Our results demonstrate a wiring design, likely engaging homotypic axonal tiling of BCs, that ensures consistency in synaptic convergence between specific BC types onto their target RGCs while enabling independent regulation of pre- and postsynaptic territory sizes and synapse number between cell pairs.
Assuntos
Células Ganglionares da Retina/metabolismo , Sinapses/metabolismo , Animais , Axônios/metabolismo , Dendritos/metabolismo , Glutamatos/metabolismo , Camundongos , Células Bipolares da Retina/metabolismo , Peixe-Zebra/metabolismoRESUMO
Understanding how neurons encode and compute information is fundamental to our study of the brain, but opportunities for hands-on experience with neurophysiological techniques on live neurons are scarce in science education. Here, we present Spikeling, an open source in silico implementation of a spiking neuron that costs £25 and mimics a wide range of neuronal behaviours for classroom education and public neuroscience outreach. Spikeling is based on an Arduino microcontroller running the computationally efficient Izhikevich model of a spiking neuron. The microcontroller is connected to input ports that simulate synaptic excitation or inhibition, to dials controlling current injection and noise levels, to a photodiode that makes Spikeling light sensitive, and to a light-emitting diode (LED) and speaker that allows spikes to be seen and heard. Output ports provide access to variables such as membrane potential for recording in experiments or digital signals that can be used to excite other connected Spikelings. These features allow for the intuitive exploration of the function of neurons and networks mimicking electrophysiological experiments. We also report our experience of using Spikeling as a teaching tool for undergraduate and graduate neuroscience education in Nigeria and the United Kingdom.
Assuntos
Potenciais de Ação/fisiologia , Modelos Neurológicos , Neurônios/fisiologia , Neurociências/educação , Neurociências/instrumentação , Animais , Relações Comunidade-Instituição , Simulação por Computador , Desenho de Equipamento , Rede Nervosa/fisiologia , Redes Neurais de Computação , Sinapses/fisiologiaRESUMO
Glutamate is the major excitatory neurotransmitter in the brain. Its release and eventual recycling are key to rapid sustained neural activity. We have paired the gfap promoter region with the glutamate reporter molecule, iGluSnFR, to drive expression in glial cells throughout the nervous system. Tg(gfap:iGluSnFR) is expressed on the glial membrane of Müller glia cells in the retina, which rapidly respond to stimulation and the release of extracellular glutamate. As glial cells are associated with most, if not all, synapses, Tg(gfap:iGluSnFR) is a novel and exciting tool to measure neuronal activity and extracellular glutamate dynamics in many regions of the nervous system.
Assuntos
Proteínas de Escherichia coli/metabolismo , Ácido Glutâmico/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Sistema Nervoso/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Peixe-Zebra/metabolismo , Animais , Animais Geneticamente Modificados/crescimento & desenvolvimento , Animais Geneticamente Modificados/metabolismo , Regulação da Expressão Gênica , Sistema Nervoso/citologia , Neuroglia/citologia , Neuroglia/metabolismo , Regiões Promotoras Genéticas , Retina/citologia , Retina/metabolismo , Peixe-Zebra/crescimento & desenvolvimentoRESUMO
In daylight, the input to the retinal circuit is provided primarily by cone photoreceptors acting as band-pass filters, but the retinal output also contains neuronal populations transmitting sustained signals. Using in vivo imaging of genetically encoded calcium reporters, we investigated the circuits that generate these sustained channels within the inner retina of zebrafish. In OFF bipolar cells, sustained transmission was found to depend on crossover inhibition from the ON pathway through GABAergic amacrine cells. In ON bipolar cells, the amplitude of low-frequency signals was regulated by glycinergic amacrine cells, while GABAergic inhibition regulated the gain of band-pass signals. We also provide the first functional description of a subset of sustained ON bipolar cells in which synaptic activity was suppressed by fluctuations at frequencies above â¼0.2 Hz. These results map out the basic circuitry by which the inner retina generates sustained visual signals and describes a new function of crossover inhibition.