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1.
Cell Rep ; 43(3): 113943, 2024 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-38483907

RESUMO

The maturation of engrams from recent to remote time points involves the recruitment of CA1 neurons projecting to the anterior cingulate cortex (CA1→ACC). Modifications of G-protein-coupled receptor pathways in CA1 astrocytes affect recent and remote recall in seemingly contradictory ways. To address this inconsistency, we manipulated these pathways in astrocytes during memory acquisition and tagged c-Fos-positive engram cells and CA1→ACC cells during recent and remote recall. The behavioral results were coupled with changes in the recruitment of CA1→ACC projection cells to the engram: Gq pathway activation in astrocytes caused enhancement of recent recall alone and was accompanied by earlier recruitment of CA1→ACC projecting cells to the engram. In contrast, Gi pathway activation in astrocytes resulted in the impairment of only remote recall, and CA1→ACC projecting cells were not recruited during remote memory. Finally, we provide a simple working model, hypothesizing that Gq and Gi pathway activation affect memory differently, by modulating the same mechanism: CA1→ACC projection.


Assuntos
Astrócitos , Memória de Longo Prazo , Memória de Longo Prazo/fisiologia , Memória/fisiologia , Rememoração Mental/fisiologia , Neurônios/fisiologia , Giro do Cíngulo/fisiologia , Hipocampo/fisiologia
2.
STAR Protoc ; 5(1): 102840, 2024 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-38280198

RESUMO

Here, we present a protocol for marking engram cells to efficiently measure reactivation levels and their projection pathways. We describe steps for genetic manipulation utilizing transgenic mice and viral infections, labeling engram cells, and a modified version of CLARITY, a tissue-clearing technique. This protocol can be adapted to various research inquiries that involve assessing the overlap of cell populations and uncovering novel long-range connectivity pathways. For complete details on the use and execution of this protocol, please refer to Refaeli et al. (2023).1.


Assuntos
Camundongos Transgênicos , Animais , Camundongos
3.
Curr Biol ; 33(18): 3942-3950.e3, 2023 09 25.
Artigo em Inglês | MEDLINE | ID: mdl-37586373

RESUMO

Remote memories play an important role in how we perceive the world, and they are rooted throughout the brain in "engrams": ensembles of cells that are formed during acquisition. Upon their reactivation, a specific memory can be recalled.1,2,3,4,5,6,7,8,9,10,11,12 Many studies have focused on the ensembles in CA1 of the hippocampus and the anterior cingulate cortex (ACC). However, the evolution of these components during systems' consolidation has not yet been comprehensively addressed.13,14,15,16 By applying transgenic approaches for ensemble identification, CLARITY, retro-AAV, and pseudo-rabies virus for circuit mapping, and chemogenetics for functional interrogation, we addressed the dynamics of recent and remote CA1 ensembles. We expected both stability (as they represent the same memory) and maturation (over time). Indeed, we found that CA1 engrams remain stable between recent and remote recalls, and the inhibition of engrams for recent recall during remote recall functionally impairs memory. We also found that new cells in the remote recall engram in the CA1 are not added randomly during maturation but differ according to their connections. First, we show in two ways that the anterograde CA1 → ACC engram cell projection grows larger. Finally, in the retrograde projections, the ACC reduces input to CA1 engram cells, whereas input from the entorhinal cortex and paraventricular nucleus of the thalamus increases. Our results shine fresh light on systems' consolidation by providing a deeper understanding of engram stability and maturation in the transition from recent to remote memory.


Assuntos
Hipocampo , Memória de Longo Prazo , Hipocampo/fisiologia , Memória de Longo Prazo/fisiologia , Rememoração Mental/fisiologia , Córtex Entorrinal , Giro do Cíngulo/fisiologia
4.
Nature ; 609(7928): 772-778, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-36045289

RESUMO

Astrocytic calcium dynamics has been implicated in the encoding of sensory information1-5, and modulation of calcium in astrocytes has been shown to affect behaviour6-10. However, longitudinal investigation of the real-time calcium activity of astrocytes in the hippocampus of awake mice is lacking. Here we used two-photon microscopy to chronically image CA1 astrocytes as mice ran in familiar or new virtual environments to obtain water rewards. We found that astrocytes exhibit persistent ramping activity towards the reward location in a familiar environment, but not in a new one. Shifting the reward location within a familiar environment also resulted in diminished ramping. After additional training, as the mice became familiar with the new context or new reward location, the ramping was re-established. Using linear decoders, we could predict the location of the mouse in a familiar environment from astrocyte activity alone. We could not do the same in a new environment, suggesting that the spatial modulation of astrocytic activity is experience dependent. Our results indicate that astrocytes can encode the expected reward location in spatial contexts, thereby extending their known computational abilities and their role in cognitive functions.


Assuntos
Astrócitos , Região CA1 Hipocampal , Recompensa , Animais , Astrócitos/fisiologia , Região CA1 Hipocampal/citologia , Região CA1 Hipocampal/fisiologia , Cálcio/metabolismo , Ingestão de Líquidos , Camundongos , Água
5.
J Vis Exp ; (181)2022 03 31.
Artigo em Inglês | MEDLINE | ID: mdl-35435896

RESUMO

Combining viral vector transduction and tissue clearing using the CLARITY method makes it possible to simultaneously investigate several types of brain cells and their interactions. Viral vector transduction enables the marking of diverse cell types in different fluorescence colors within the same tissue. Cells can be identified genetically by activity or projection. Using a modified CLARITY protocol, the potential sample size of astrocytes and neurons has grown by 2-3 orders of magnitude. The use of CLARITY allows the imaging of complete astrocytes, which are too large to fit in their entirety in slices, and the examination of the somata with all their processes. In addition, it provides the opportunity to investigate the spatial interaction between astrocytes and different neuronal cell types, namely, the number of pyramidal neurons in each astrocytic domain or the proximity between astrocytes and specific inhibitory neuron populations. This paper describes, in detail, how these methods are to be applied.


Assuntos
Astrócitos , Neurônios , Astrócitos/metabolismo , Encéfalo/metabolismo , Neurônios/fisiologia , Células Piramidais
6.
Neuron ; 110(3): 363-365, 2022 02 02.
Artigo em Inglês | MEDLINE | ID: mdl-35114108

RESUMO

This NeuroView is intended for graduate students who are not sure how to choose or what to expect from a mentor as well as mentors who are uncertain what to give mentees. Two principal investigators and a current mentee will share their perspectives on this bidirectional relationship.


Assuntos
Tutoria , Mentores , Humanos , Avaliação de Programas e Projetos de Saúde , Estudantes
7.
Glia ; 69(10): 2378-2390, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34117643

RESUMO

The mounting evidence for the involvement of astrocytes in neuronal circuits function and behavior stands in stark contrast to the lack of detailed anatomical description of these cells and the neurons in their domains. To fill this void, we imaged >30,000 astrocytes in hippocampi made transparent by CLARITY, and determined the elaborate structure, distribution, and neuronal content of astrocytic domains. First, we characterized the spatial distribution of >19,000 astrocytes across CA1 lamina, and analyzed the morphology of thousands of reconstructed domains. We then determined the excitatory somatic content of CA1 astrocytes, and measured the distance between inhibitory neuronal somata to the nearest astrocyte soma. We find that on average, there are almost 14 pyramidal neurons per domain in the CA1, increasing toward the pyramidal layer midline, compared to only five excitatory neurons per domain in the amygdala. Finally, we discovered that somatostatin neurons are found in close proximity to astrocytes, compared to parvalbumin and VIP inhibitory neurons. This work provides a comprehensive large-scale quantitative foundation for studying neuron-astrocyte interactions.


Assuntos
Astrócitos , Hipocampo , Neurônios/fisiologia , Células Piramidais/fisiologia
8.
Curr Opin Neurobiol ; 67: 131-137, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33260057

RESUMO

For decades, the study of memory has been neuron-centric, yet neurons do not function in isolation. Today we know that neuronal activity is modulated by the environment within which it occurs, and is subject to modulation by different types of glial cells. In this review we summarize recent findings on the functional roles of astrocytes and oligodendrocytes, two major types of glia cells in the adult brain, in memory formation and its cellular underpinnings across multiple time points. We will discuss the different methods that are being used to investigate the astrocytic and oligodendroglial involvement in memory. We shall focus on chemogenetics and optogenetics, which support genetically specificity and high spatiotemporal resolution, attributes that are particularly well suited to the investigation of the contribution of unique cell types at the different stages of memory formation.


Assuntos
Astrócitos , Memória , Neuroglia , Neurônios , Oligodendroglia
9.
Nat Neurosci ; 23(10): 1229-1239, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32747787

RESUMO

Remote memories depend on coordinated activity in the hippocampus and frontal cortices, but the timeline of these interactions is debated. Astrocytes sense and modify neuronal activity, but their role in remote memory is scarcely explored. We expressed the Gi-coupled designer receptor hM4Di in CA1 astrocytes and discovered that astrocytic manipulation during learning specifically impaired remote, but not recent, memory recall and decreased activity in the anterior cingulate cortex (ACC) during retrieval. We revealed massive recruitment of ACC-projecting CA1 neurons during memory acquisition, which was accompanied by the activation of ACC neurons. Astrocytic Gi activation disrupted CA3 to CA1 communication in vivo and reduced the downstream response in the ACC. In behaving mice, it induced a projection-specific inhibition of CA1-to-ACC neurons during learning, which consequently prevented ACC recruitment. Finally, direct inhibition of CA1-to-ACC-projecting neurons spared recent and impaired remote memory. Our findings suggest that remote memory acquisition involves projection-specific functions of astrocytes in regulating CA1-to-ACC neuronal communication.


Assuntos
Astrócitos/fisiologia , Giro do Cíngulo/fisiologia , Hipocampo/fisiologia , Aprendizagem/fisiologia , Memória/fisiologia , Neurônios/fisiologia , Animais , Condicionamento Clássico/fisiologia , Medo/fisiologia , Masculino , Rememoração Mental/fisiologia , Camundongos Endogâmicos C57BL , Vias Neurais/fisiologia
10.
Neuron ; 105(1): 9-11, 2020 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-31951529

RESUMO

Adult oligodendrogenesis is regulated by neuronal activity and learning. Can it affect memory processes? In this issue of Neuron, Steadman et al. (2020) found that newly generated oligodendrocytes are crucial for memory acquisition and consolidation and required for the neuronal coupling between brain regions known to be involved in memory.


Assuntos
Consolidação da Memória , Animais , Aprendizagem , Memória , Camundongos , Neuroglia , Oligodendroglia
11.
Glia ; 68(9): 1692-1728, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-31958188

RESUMO

Development, physiological functions, and pathologies of the brain depend on tight interactions between neurons and different types of glial cells, such as astrocytes, microglia, oligodendrocytes, and oligodendrocyte precursor cells. Assessing the relative contribution of different glial cell types is required for the full understanding of brain function and dysfunction. Over the recent years, several technological breakthroughs were achieved, allowing "glio-scientists" to address new challenging biological questions. These technical developments make it possible to study the roles of specific cell types with medium or high-content workflows and perform fine analysis of their mutual interactions in a preserved environment. This review illustrates the potency of several cutting-edge experimental approaches (advanced cell cultures, induced pluripotent stem cell (iPSC)-derived human glial cells, viral vectors, in situ glia imaging, opto- and chemogenetic approaches, and high-content molecular analysis) to unravel the role of glial cells in specific brain functions or diseases. It also illustrates the translation of some techniques to the clinics, to monitor glial cells in patients, through specific brain imaging methods. The advantages, pitfalls, and future developments are discussed for each technique, and selected examples are provided to illustrate how specific "gliobiological" questions can now be tackled.


Assuntos
Astrócitos , Neuroglia , Humanos , Microglia , Neurônios , Oligodendroglia
12.
Curr Biol ; 28(17): 2752-2762.e7, 2018 09 10.
Artigo em Inglês | MEDLINE | ID: mdl-30122531

RESUMO

A barrage of information constantly assaults our senses, of which only a fraction is relevant at any given point in time. However, the neural circuitry supporting the suppression of irrelevant sensory distractors is not completely understood. The claustrum, a circuit hub with vast cortical connectivity, is an intriguing brain structure, whose restrictive anatomy, thin and elongated, has precluded functional investigation. Here, we describe the use of Egr2-CRE mice to access genetically defined claustral neurons. Utilizing conditional viruses for anterograde axonal labeling and retrograde trans-synaptic tracing, we validated this transgenic model for accessing the claustrum and extended the known repertoire of claustral input/output connectivity. Addressing the function of the claustrum, we inactivated CLEgr2+ neurons, chronically as well as acutely, in mice performing an automated two-alternative forced-choice behavioral task. Strikingly, inhibition of CLEgr2+ neurons did not significantly impact task performance under varying delay times and cue durations, but revealed a selective role for the claustrum in supporting performance in the presence of an irrelevant auditory distractor. Further investigation of behavior, in the naturalistic maternal pup-retrieval task, replicated the result of sensitization to an auditory distractor following inhibition of CLEgr2+ neurons. Initiating investigation into the underlying mechanism, we found that activation of CLEgr2+ neurons modulated cortical sensory processing, suppressing tone representation in the auditory cortex. This functional study, utilizing selective genetic access, implicates the claustrum in supporting resilience to distraction, a fundamental aspect of attention.


Assuntos
Atenção/fisiologia , Gânglios da Base/fisiologia , Neurônios/fisiologia , Animais , Comportamento Animal/fisiologia , Proteína 2 de Resposta de Crescimento Precoce/genética , Proteína 2 de Resposta de Crescimento Precoce/metabolismo , Feminino , Técnicas de Introdução de Genes , Integrases/genética , Integrases/metabolismo , Camundongos , Camundongos Endogâmicos , Vias Neurais/fisiologia
13.
Cell ; 174(1): 59-71.e14, 2018 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-29804835

RESUMO

Astrocytes respond to neuronal activity and were shown to be necessary for plasticity and memory. To test whether astrocytic activity is also sufficient to generate synaptic potentiation and enhance memory, we expressed the Gq-coupled receptor hM3Dq in CA1 astrocytes, allowing their activation by a designer drug. We discovered that astrocytic activation is not only necessary for synaptic plasticity, but also sufficient to induce NMDA-dependent de novo long-term potentiation in the hippocampus that persisted after astrocytic activation ceased. In vivo, astrocytic activation enhanced memory allocation; i.e., it increased neuronal activity in a task-specific way only when coupled with learning, but not in home-caged mice. Furthermore, astrocytic activation using either a chemogenetic or an optogenetic tool during acquisition resulted in memory recall enhancement on the following day. Conversely, directly increasing neuronal activity resulted in dramatic memory impairment. Our findings that astrocytes induce plasticity and enhance memory may have important clinical implications for cognitive augmentation treatments.


Assuntos
Potenciação de Longa Duração , Memória , Neurônios/metabolismo , Animais , Astrócitos/citologia , Astrócitos/efeitos dos fármacos , Astrócitos/metabolismo , Cálcio/metabolismo , Clozapina/análogos & derivados , Clozapina/farmacologia , Subunidades alfa Gq-G11 de Proteínas de Ligação ao GTP/genética , Subunidades alfa Gq-G11 de Proteínas de Ligação ao GTP/metabolismo , Hipocampo/citologia , Potenciação de Longa Duração/efeitos dos fármacos , Masculino , Memória/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , N-Metilaspartato/farmacologia , Neurônios/efeitos dos fármacos , Optogenética , Técnicas de Patch-Clamp , Proteínas Proto-Oncogênicas c-fos/metabolismo , Estresse Psicológico , Potenciais Sinápticos/efeitos dos fármacos
14.
Brain Res Bull ; 141: 35-43, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-28939475

RESUMO

Remote memories, weeks to decades long, are usually the ones most important to the organism, as the longevity of a memory is tightly connected to its significance. Retrograde amnesia studies in human patients as well as lesions and immediate early gene expression investigation in animal models, suggested that the hippocampus has a time dependent role in memory consolidation. Namely, that as a memory matures it becomes independent of the hippocampus and instead depends on extra-hippocampal areas. However, accumulating evidence implies that this temporal segregation is not as rigid as originally proposed. In this review we will focus on the integration of new methods, such as chemogenetics, optogenetics and calcium imaging, which enable genetic specificity as well as high temporal and spatial resolution. Using these methods, recent studies have started to resolve the inconsistencies of past findings by observing and manipulating neural ensembles in different brain regions. We then discuss how these techniques can be applied to investigate the cellular underpinnings of memory across multiple time points, and employed to study the contribution of various cell types to remote memory.


Assuntos
Encéfalo/citologia , Encéfalo/diagnóstico por imagem , Técnicas Genéticas , Memória de Longo Prazo/fisiologia , Memória de Curto Prazo/fisiologia , Animais , Encéfalo/fisiologia , Encéfalo/fisiopatologia , Humanos , Imagens com Corantes Sensíveis à Voltagem
15.
Neuroscience ; 370: 14-26, 2018 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-28571720

RESUMO

Astrocytes have been generally believed to perform mainly homeostatic and supportive functions for neurons in the central nervous system. Recently, a growing body of evidence suggests previously unrecognized and surprising functions for astrocytes, including regulation of synaptic formation, transmission and plasticity, all of which are considered as the infrastructure for information processing and memory formation and stabilization. This review discusses the involvement of astrocytes in memory functions and the possible mechanisms that may underlie it. We review the important breakthroughs obtained in this field, as well as some of the controversies that arose from the past difficulty to manipulate these cells in a cell type-specific and non-invasive manner. Finally, we present new research avenues based on the advanced tools becoming available in recent years: optogenetics and chemogenetics, and the potential ways in which these tools may further illuminate the role of astrocytes in memory processes.


Assuntos
Astrócitos/fisiologia , Memória/fisiologia , Animais , Humanos , Plasticidade Neuronal/fisiologia
16.
Cereb Cortex ; 27(6): 3457-3470, 2017 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-28407141

RESUMO

Hippocampus, a temporal lobe structure involved in learning and memory, receives information from all sensory modalities. Despite extensive research on the role of sensory experience in cortical map plasticity, little is known about whether and how sensory experience regulates functioning of the hippocampal circuits. Here, we show that 9 ± 2 days of whisker deprivation during early mouse development depresses activity of CA3 pyramidal neurons by several principal mechanisms: decrease in release probability, increase in the fraction of silent synapses, and reduction in intrinsic excitability. As a result of deprivation-induced presynaptic inhibition, CA3-CA1 synaptic facilitation was augmented at high frequencies, shifting filtering properties of synapses. The changes in the AMPA-mediated synaptic transmission were accompanied by an increase in NR2B-containing NMDA receptors and a reduction in the AMPA/NMDA ratio. The observed reconfiguration of the CA3-CA1 connections may represent a homeostatic adaptation to augmentation in synaptic activity during the initial deprivation phase. In adult mice, tactile disuse diminished intrinsic excitability without altering synaptic facilitation. We suggest that sensory experience regulates computations performed by the hippocampus by tuning its synaptic and intrinsic characteristics.


Assuntos
Potenciais Pós-Sinápticos Excitadores/fisiologia , Hipocampo/fisiologia , Rede Nervosa/fisiologia , Neurônios/fisiologia , Privação Sensorial/fisiologia , Transmissão Sináptica/fisiologia , Fatores Etários , Animais , Animais Recém-Nascidos , Corticosterona/sangue , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Comportamento Exploratório/fisiologia , Técnicas In Vitro , Aprendizagem em Labirinto/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , N-Metilaspartato/metabolismo , Rede Nervosa/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Neurotransmissores/farmacologia , Receptores de N-Metil-D-Aspartato/metabolismo , Transmissão Sináptica/efeitos dos fármacos , Vibrissas/inervação , Ácido alfa-Amino-3-hidroxi-5-metil-4-isoxazol Propiônico/metabolismo
17.
Cell Transplant ; 25(7): 1371-80, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26132738

RESUMO

Compelling evidence suggests that transplantation of neural stem cells (NSCs) from multiple sources ameliorates motor deficits after stroke. However, it is currently unknown to what extent the electrophysiological activity of grafted NSC progeny participates in the improvement of motor deficits and whether excitatory phenotypes of the grafted cells are beneficial or deleterious to sensorimotor performances. To address this question, we used optogenetic tools to drive the excitatory outputs of the grafted NSCs and assess the impact on local circuitry and sensorimotor performance. We genetically engineered NSCs to express the Channelrhodopsin-2 (ChR2), a light-gated cation channel that evokes neuronal depolarization and initiation of action potentials with precise temporal control to light stimulation. To test the function of these cells in a stroke model, rats were subjected to an ischemic stroke and grafted with ChR2-NSCs. The grafted NSCs identified with a human-specific nuclear marker survived in the peri-infarct tissue and coexpressed the ChR2 transgene with the neuronal markers TuJ1 and NeuN. Gene expression analysis in stimulated versus vehicle-treated animals showed a differential upregulation of transcripts involved in neurotransmission, neuronal differentiation, regeneration, axonal guidance, and synaptic plasticity. Interestingly, genes involved in the inflammatory response were significantly downregulated. Behavioral analysis demonstrated that chronic optogenetic stimulation of the ChR2-NSCs enhanced forelimb use on the stroke-affected side and motor activity in an open field test. Together these data suggest that excitatory stimulation of grafted NSCs elicits beneficial effects in experimental stroke model through cell replacement and non-cell replacement, anti-inflammatory/neurotrophic effects.


Assuntos
Regulação para Baixo , Células-Tronco Neurais/transplante , Optogenética/métodos , Acidente Vascular Cerebral/terapia , Transmissão Sináptica , Animais , Separação Celular , Modelos Animais de Doenças , Perfilação da Expressão Gênica , Células-Tronco Embrionárias Humanas/citologia , Humanos , Inflamação/complicações , Inflamação/genética , Inflamação/terapia , Masculino , Neostriado/metabolismo , Células-Tronco Neurais/citologia , Análise de Sequência com Séries de Oligonucleotídeos , Ratos Sprague-Dawley , Rodopsina/genética , Acidente Vascular Cerebral/complicações , Acidente Vascular Cerebral/genética , Transdução Genética , Transgenes
18.
Trends Neurosci ; 37(9): 511-22, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25022518

RESUMO

Over the past 5 years, the incorporation of optogenetics into the study of memory has resulted in a tremendous leap in this field, initiating a revolution in our understanding of the networks underlying cognitive processes. This review will present recent breakthroughs in which optogenetics was applied to illuminate, both literally and figuratively, memory research, and describe the technical approach, together with the opportunities it offers. Specifically, a large body of literature has been generated, setting the foundation for deciphering the spatiotemporal organization of hippocampal-based memory processing and its underlying mechanisms, as well as the contribution of cortical and amygdalar regions to cognition.


Assuntos
Encéfalo/fisiologia , Memória/fisiologia , Optogenética/métodos , Animais , Humanos , Aprendizagem/fisiologia , Vias Neurais/fisiologia
19.
Nat Methods ; 9(12): 1171-9, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23169303

RESUMO

Optogenetics with microbial opsin genes has enabled high-speed control of genetically specified cell populations in intact tissue. However, it remains a challenge to independently control subsets of cells within the genetically targeted population. Although spatially precise excitation of target molecules can be achieved using two-photon laser-scanning microscopy (TPLSM) hardware, the integration of two-photon excitation with optogenetics has thus far required specialized equipment or scanning and has not yet been widely adopted. Here we take a complementary approach, developing opsins with custom kinetic, expression and spectral properties uniquely suited to scan times typical of the raster approach that is ubiquitous in TPLSMlaboratories. We use a range of culture, slice and mammalian in vivo preparations to demonstrate the versatility of this toolbox, and we quantitatively map parameter space for fast excitation, inhibition and bistable control. Together these advances may help enable broad adoption of integrated optogenetic and TPLSMtechnologies across experimental fields and systems.


Assuntos
Microscopia Confocal/instrumentação , Neurônios/fisiologia , Opsinas/genética , Optogenética , Animais , Células Cultivadas , Desenho de Equipamento , Masculino , Potenciais da Membrana/fisiologia , Camundongos , Fótons , Transfecção
20.
Nat Neurosci ; 15(1): 163-70, 2011 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-22138641

RESUMO

Recent advances in optogenetics have improved the precision with which defined circuit elements can be controlled optically in freely moving mammals; in particular, recombinase-dependent opsin viruses, used with a growing pool of transgenic mice expressing recombinases, allow manipulation of specific cell types. However, although optogenetic control has allowed neural circuits to be manipulated in increasingly powerful ways, combining optogenetic stimulation with simultaneous multichannel electrophysiological readout of isolated units in freely moving mice remains a challenge. We designed and validated the optetrode, a device that allows for colocalized multi-tetrode electrophysiological recording and optical stimulation in freely moving mice. Optetrode manufacture employs a unique optical fiber-centric coaxial design approach that yields a lightweight (2 g), compact and robust device that is suitable for behaving mice. This low-cost device is easy to construct (2.5 h to build without specialized equipment). We found that the drive design produced stable high-quality recordings and continued to do so for at least 6 weeks following implantation. We validated the optetrode by quantifying, for the first time, the response of cells in the medial prefrontal cortex to local optical excitation and inhibition, probing multiple different genetically defined classes of cells in the mouse during open field exploration.


Assuntos
Desenho de Equipamento , Técnicas Genéticas/instrumentação , Animais , Camundongos
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