Fast and sensitive GCaMP calcium indicators for imaging neural populations

Nature, Год журнала: 2023, Номер 615(7954), С. 884 - 891

Опубликована: Март 15, 2023

Abstract Calcium imaging with protein-based indicators 1,2 is widely used to follow neural activity in intact nervous systems, but current protein sensors report at timescales much slower than electrical signalling and are limited by trade-offs between sensitivity kinetics. Here we large-scale screening structure-guided mutagenesis develop optimize several fast sensitive GCaMP-type 3–8 . The resulting ‘jGCaMP8’ sensors, based on the calcium-binding calmodulin a fragment of endothelial nitric oxide synthase, have ultra-fast kinetics (half-rise times 2 ms) highest for reported calcium sensor. jGCaMP8 will allow tracking large populations neurons relevant computation.

Язык: Английский

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Dense connectomic reconstruction in layer 4 of the somatosensory cortex

Science, Год журнала: 2019, Номер 366(6469)

Опубликована: Окт. 24, 2019

The dense circuit structure of mammalian cerebral cortex is still unknown. With developments in three-dimensional electron microscopy, the imaging sizable volumes neuropil has become possible, but reconstruction connectomes limiting step. We reconstructed a volume ~500,000 cubic micrometers from layer 4 mouse barrel cortex, ~300 times larger than previous reconstructions cortex. connectomic data allowed extraction inhibitory and excitatory neuron subtypes that were not predictable geometric information. quantified imprints consistent with Hebbian synaptic weight adaptation, which yielded upper bounds for fraction saturated long-term potentiation. These establish an approach locally phenotyping neuronal circuitry

Язык: Английский

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Cellular Mechanisms of Conscious Processing

Trends in Cognitive Sciences, Год журнала: 2020, Номер 24(10), С. 814 - 825

Опубликована: Авг. 24, 2020

Recent breakthroughs in neurobiology indicate that the time is ripe to understand how cellular-level mechanisms are related conscious experience. Here, we highlight biophysical properties of pyramidal cells, which allow them act as gates control evolution global activation patterns. In states, this cellular mechanism enables complex sustained dynamics within thalamocortical system, whereas during unconscious such signal propagation prohibited. We suggest hallmark processing flexible integration bottom-up and top-down data streams at level. This provides foundation for Dendritic Information Theory, a novel neurobiological theory consciousness.

Язык: Английский

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Spatiotemporal constraints on optogenetic inactivation in cortical circuits

eLife, Год журнала: 2019, Номер 8

Опубликована: Ноя. 18, 2019

Optogenetics allows manipulations of genetically and spatially defined neuronal populations with excellent temporal control. However, neurons are coupled other over multiple length scales, the effects localized thus spread beyond targeted neurons. We benchmarked several optogenetic methods to inactivate small regions neocortex. Optogenetic excitation GABAergic produced more effective inactivation than light-gated ion pumps. Transgenic mice expressing light-dependent chloride channel GtACR1 most potent inactivation. Generally, substantially photostimulation light, caused by strong coupling between cortical Over some range light intensity, inhibitory reduced activity in these neurons, together pyramidal a signature inhibition-stabilized neural networks ('paradoxical effect'). The offset was followed rebound dose-dependent manner, limiting resolution. Our data offer guidance for design vivo optogenetics experiments.

Язык: Английский

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Circuitry Underlying Experience-Dependent Plasticity in the Mouse Visual System

Neuron, Год журнала: 2020, Номер 106(1), С. 21 - 36

Опубликована: Апрель 1, 2020

Since the discovery of ocular dominance plasticity, neuroscientists have understood that changes in visual experience during a discrete developmental time, critical period, trigger robust cortex. State-of-the-art tools used to probe connectivity with cell-type-specific resolution expanded understanding circuit underlying experience-dependent plasticity. Here, we review circuitry mouse, describing projections from retina thalamus, between thalamus and cortex, within We discuss how development leads precise identify synaptic loci, which can be altered by activity or experience. Plasticity extends features beyond dominance, involving subcortical cortical regions, connections inhibitory interneurons. Experience-dependent plasticity contributes alignment networks spanning Disruption this may underlie aberrant sensory processing some neurodevelopmental disorders. Vision is so inseparable our conception world "seeing believing." Sight crucial recognize friends, find savory food, plan movement. While aspects system form utero, circuits significant spontaneous activity- reorganization early life. Developmental refinement perturbations small errors, imparting uniqueness based on without an exhaustive molecular code specifying each connection. occur at specific time points. The best-known sensitive periods (SPs) (CPs). SPs are windows when neuronal exhibit modification. Such easily evoked greater magnitude SP, but smaller extent other ages. CPs subset SPs, distinguished strict temporal onset closure fact period required for subsequent normal function (Voss, 2013Voss P. Sensitive deprivation.Front. Psychol. 2013; 4: 664Crossref PubMed Scopus (25) Google Scholar). This change not reversible except CP. CP was described Hubel Wiesel. In binocular responsiveness input deprived eye attenuated weeks monocular deprivation (MD). (OD), called (ODP), only occurs development. Moreover, include reduction acuity eye, amblyopia. ODP reversed CP, recovery possible once closes. Growing evidence suggests entirely extinguished closure. Substantial adult outside activated, often stimulus paradigms distinct those effective (Espinosa Stryker, 2012Espinosa J.S. Stryker M.P. Development primary cortex.Neuron. 2012; 75: 230-249Abstract Full Text PDF (294) Scholar, Fong et al., 2016Fong M.F. Mitchell D.E. Duffy K.R. Bear Rapid effects enabled temporary inactivation retinas.Proc Natl Acad Sci U S A. 2016; 113: 14139-14144Crossref (14) Hensch Quinlan, 2018Hensch T.K. Quinlan E.M. Critical amblyopia.Vis. Neurosci. 2018; 35: E014PubMed Sato 2008Sato M. Distinctive plasticity.J. 2008; 28: 10278-10286Crossref (153) Thus, perhaps more accurately defined as occurring SP. ambiguity highlights difficulty distinguishing especially monitor developed. Different loci along pathway vary their sensitivity (Kang 2013Kang E. Durand S. LeBlanc J.J. Chen C. Fagiolini Visual absence experience.J. 33: 17789-17796Crossref (42) Scholar), one synapse enduring effect overall because its influence downstream synapses irreversibly. Homeostatic responses also activated compensate synapse, another (Whitt 2014Whitt J.L. Petrus Lee H.K. homeostatic neocortex.Neuropharmacology. 2014; 78: 45-54Crossref (39) end, indelibly Therefore, purpose review, define window there response followed clear diminishing same stimuli. Wiesel, has served model studying Changes following MD measured comparing (by single unit recordings, [VEPs], vivo imaging) stimulation eye. These straightforward measurements made canonical experimental system. More recently, enabling measurement such orientation preference suggest many show occasionally different rules. implies traditionally circuitry, thalamic relay cells, developmentally regulated unrelated OD shifts. Considerable effort been devoted basis (Fagiolini 2004Fagiolini Fritschy J.M. Löw K. Möhler H. Rudolph U. Specific GABAA plasticity.Science. 2004; 303: 1681-1683Crossref (332) means label, monitor, manipulate these components independently understand basic where results changes. mouse powerful model. Despite zone lower 1994Fagiolini Pizzorusso T. Berardi N. Domenici L. Maffei Functional postnatal rat cortex role experience: dark rearing deprivation.Vision Res. 1994; 34: 709-720Crossref (491) mice similar primates cats. clearly day 19 (P19) P32 (Gordon 1996Gordon J.A. mouse.J. 1996; 16: 3274-3286Crossref Mice offer possibility genetic access neuron populations ODP, including subsets excitatory (Gerfen 2013Gerfen C.R. Paletzki R. Heintz GENSAT BAC cre-recombinase driver lines study functional organization cerebral basal ganglia circuits.Neuron. 80: 1368-1383Abstract (242) Scholar) neurons (Taniguchi 2011Taniguchi He Wu Kim Paik Sugino Kvitsiani D. Fu Y. Lu J. Lin al.A resource Cre targeting GABAergic 2011; 71: 995-1013Abstract (822) approaches allow targeted recording, optical manipulation (Madisen 2012Madisen Mao Koch Zhuo Berenyi Fujisawa Hsu Y.W. Garcia 3rd, A.J. Gu X. Zanella toolbox Cre-dependent optogenetic transgenic light-induced activation silencing.Nat. 15: 793-802Crossref (616) calcium indicators (Chen 2013Chen T.W. Wardill T.J. Sun Pulver S.R. Renninger S.L. Baohan Schreiter E.R. Kerr R.A. Orger M.B. Jayaraman V. al.Ultrasensitive fluorescent proteins imaging activity.Nature. 499: 295-300Crossref (2349) advances focusing across species, mechanisms reviewed elsewhere (Hensch Kiorpes, 2015Kiorpes primates: neural periods.Dev. Neurobiol. 2015; 1080-1090Crossref refer work areas, extrapolating Studies revealed apply than simply OD. With variety manipulations quantifying receptive field (RF) properties, orientation, luminance, spatial frequency tuning, findings rules differ ODP. studies demonstrate To instantiated, detailed map needed, long-range major brain structures (Figures 1 2) cell types 3 4). true neocortex, contains types, organized layers, multiple per layer. A near-complete census (Sugino 2019Sugino Clark Schulmann Shima Wang Hunt D.L. Hooks B.M. Tränkner Chandrashekar Picard al.Mapping transcriptional diversity genetically anatomically brain.eLife. 2019; 8: e38619Crossref (3) Tasic 2018Tasic B. Yao Z. Graybuck L.T. Smith K.A. Nguyen T.N. Bertagnolli Goldy Garren Economo M.N. Viswanathan al.Shared transcriptomic neocortical areas.Nature. 563: 72-78Crossref (167) promises full range subtypes. provides parts list local circuit. Given profoundly plasticity? Our aim place order context, assessing whether hierarchy earlier way nodes thalamocortical (TC) afferents compared intracortical L4 L2/3.Figure 2Thalamocortical OrganizationShow caption(A) includes dorsal ventral subdivisions (dLGN vLGN, blue purple). Core shell areas dLGN receive inputs RGC types. Higher-order LP (red).(B) Thalamic axons V1 arborize laminae.View Large Image Figure ViewerDownload Hi-res image Download (PPT)Figure 3Inhibitory Excitatory Local Connectivity CortexShow Inhibitory interneurons laminar distribution 5-HT3AR+ concentrated L1 L2/3, VIP+ PV+ SST+ present all (after 2010Lee Hjerling-Leffler Zagha Fishell G. Rudy largest group superficial expresses ionotropic serotonin receptors.J. 2010; 30: 16796-16808Crossref (323) Xu 2010Xu Roby K.D. Callaway Immunochemical characterization neurons: three chemically classes cells.J. Comp. Neurol. 518: 389-404Crossref (256) Subtypes highlighted table right.(B) bipolar disinhibit specifically inhibiting neurons. cells tonically inhibit pyramidal (Pyr) apical dendrites. Tonic inhibition released become active (Pfeffer 2013Pfeffer C.K. Xue Huang Z.J. Scanziani Inhibition cortex: logic molecularly interneurons.Nat. 1068-1076Crossref (527) Input VIP (black magenta arrows) areas. L4, less abundant.(C D) Major granular supragranular given layer nearby layers (Xu 2016Xu Olivas N.D. Ikrar Peng Holmes T.C. Nie Q. Shi Primary shows laminar-specific balanced connectivity.J. Physiol. 594: 1891-1910Crossref Ascending (left) descending (right), thicknesses proportional connection strength. Laminae shown red. Strength illustration (C) matrix (D), pre- postsynaptic labeled.(E) L2/3 L4.View 4Developmental Cortical Near Period (CP)Show For illustrated. illustrated black. (blue), (burgundy), (green) color coded, neuromodulatory acetylcholine (purple). TC age annotated bottom. interneuron precede (Jiang 2010Jiang Sohya Sarihi Yanagawa Tsumoto Laminar-specific maturation transmission susceptibility related endocannabinoid cortex.J. 14261-14272Crossref (33) PV correlated matures (Cossell 2015Cossell Iacaruso Muir D.R. Houlton Sader E.N. Ko Hofer S.B. Mrsic-Flogel T.D. strength cortex.Nature. 399-403Crossref (191) 2013Ko Cossell Baragli Antolik Clopath emergence microcircuits 496: 96-100Crossref (198) neurons, initially strong, grows weaker loses cholinergic modulation (Yaeger 2019Yaeger C.E. Ringach Trachtenberg J.T. Neuromodulatory control localized dendritic spiking 567: 100-104Crossref Scholar).(B) Mature summarized.View (PPT) (A) (red). (B) laminae. right. abundant. (C labeled. (E) L4. summarized. mammals begins retina, retinal ganglion (RGC) target number superior colliculus (SC), lateral geniculate nucleus (dLGN), suprachiasmatic (SCN; Figures 1A–1C). Blue red figure illustrate corresponding ipsilateral contralateral eyes, eye-specific segregation dLGN. projection rounded patch surrounded larger area input. Some overlap exists (Figure 1B; Howarth 2014Howarth Walmsley Brown T.M. Binocular integration nuclei.Curr. Biol. 24: 1241-1247Abstract subdivided into core region, projecting principally 4 (L4) (V1, VISp reference atlas; Kuan 2015Kuan Li Lau Feng Bernard Sunkin S.M. Zeng Dang Hawrylycz Ng Neuroinformatics Allen Mouse Brain Atlas.Methods. 73: 4-17Crossref (72) largely targets (Cruz-Martín 2014Cruz-Martín El-Danaf R.N. Osakada F. Sriram Dhande O.S. P.L. Ghosh Huberman A.D. dedicated links direction-selective 507: 358-361Crossref (151) posterior (LP) (similar pulvinar primates), projects strongly L1, deeper L5A. structure output (Bennett 2019Bennett Gale S.D. Garrett M.E. Newton M.L. Murphy G.J. Olsen Higher-Order Circuits Channel Parallel Streams Information Mice.Neuron. 102: 477-492.e5Abstract (23) Two intrinsic interneurons, former outnumber latter ∼4:1. Intrinsic make dendrodendritic axodendritic onto (triads; Bickford 2010Bickford Slusarczyk Dilger E.K. Krahe T.E. Kucuk Guido W. Synaptic nucleus.J. 622-635Crossref (52) Morgan 2016Morgan Berger Wetzel A.W. Lichtman J.W. Fuzzy Logic Network Thalamus.Cell. 165: 192-206Abstract (88) Rafols Valverde, 1973Rafols Valverde mouse. Golgi electron microscopic study.J. 1973; 150: 303-332Crossref two corticothalamic (CT) well reticular (TRNs). project sending collaterals TRN. Interneurons parvalbumin positive (PV+), whereas PV−, although complete (Kalish 2018Kalish B.T. Cheadle Hrvatin Nagy M.A. Rivera Crow Gillis Kirchner Greenberg Single-cell transcriptomics developing reveals insights assembly refinement.Proc. Natl. Acad. Sci. USA. 115: E1051-E1060Crossref (17) CT inputs, turn, send TRNs. integrates, modifies, relays information passing V1. Experience-Dependent Thalamus. cat, much (Wiesel Hubel, 1963aWiesel D.H. striate kittens vision eye.J. Neurophysiol. 1963; 26: 1003-1017Crossref (1751) shrinkage somata dLGN, little (Sherman Spear, 1982Sherman Spear P.D. Organization pathways visually cats.Physiol. Rev. 1982; 62: 738-855Crossref (493) Wiesel 1963bWiesel Effects Deprivation Morphology Physiology Cells Cats Lateral Geniculate Body.J. 978-993Crossref (797) (Guzik-Kornacka 2016Guzik-Kornacka van der Bourg Vajda Joly Christ Schwab Pernet Nogo-A deletion increases optokinetic system.Brain Struct. Funct. 221: 317-329Crossref (10) led view predominantly phenomenon. contrast uncovered previously recognized. traditional divides territories, region. However, recordings dorsomedial responsive stimulation, indicating convergent both eyes (Howarth extend dendrites (Krahe 2011Krahe Henderson S.C. Morphologically regional preferences 31: 17437-17448Crossref (57) below (∼P18–P32) shifts relative depends receptor α1 subunit (Sommeijer 2017Sommeijer J.P. Ahmadlou Saiepour M.H. Seignette Min Heimel Levelt C.N. regulates critical-period thalamus.Nat. 2017; 20: 1715-1721Crossref (15) Future needed reveal potential onsets opening reversible. Notably, boutons terminating MD, even (Jaepel 2017Jaepel Hübener Bonhoeffer Rose mice.Nat. 1708-1714Crossref Bonhoeffe

Язык: Английский

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In vivo direct imaging of neuronal activity at high temporospatial resolution

Science, Год журнала: 2022, Номер 378(6616), С. 160 - 168

Опубликована: Окт. 13, 2022

There has been a long-standing demand for noninvasive neuroimaging methods that can detect neuronal activity at both high temporal and spatial resolution. We present two-dimensional fast line-scan approach enables direct imaging of with millisecond precision while retaining the resolution magnetic resonance (MRI). This was demonstrated through in vivo mouse brain 9.4 tesla during electrical whisker-pad stimulation. In spike recording optogenetics confirmed correlation observed MRI signal neural activity. It also captured sequential laminar-specific propagation along thalamocortical pathway. high-resolution, will open up new avenues science by providing deeper understanding brain's functional organization, including temporospatial dynamics networks.

Язык: Английский

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Cell-type-specific inhibitory circuitry from a connectomic census of mouse visual cortex

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2023, Номер unknown

Опубликована: Янв. 24, 2023

Mammalian cortex features a vast diversity of neuronal cell types, each with characteristic anatomical, molecular and functional properties. Synaptic connectivity powerfully shapes how type participates in the cortical circuit, but mapping rules at resolution distinct types remains difficult. Here, we used millimeter-scale volumetric electron microscopy 1 to investigate all inhibitory neurons across densely-segmented population 1352 cells spanning layers mouse visual cortex, producing wiring diagram connections more than 70,000 synapses. Taking data-driven approach inspired by classical neuroanatomy, classified based on relative targeting dendritic compartments other developed novel classification excitatory morphological synaptic input The between revealed class disinhibitory specialist basket cells, addition familiar subclasses. Analysis onto found widespread specificity, many interneurons exhibiting differential certain subpopulations spatially intermingled potential targets. Inhibitory was organized into “motif groups,” diverse sets that collectively target both perisomatic same Collectively, our analysis identified new organizing principles for inhibition will serve as foundation linking modern multimodal atlases diagram.

Язык: Английский

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Cholinergic and noradrenergic axonal activity contains a behavioral-state signal that is coordinated across the dorsal cortex

eLife, Год журнала: 2023, Номер 12

Опубликована: Апрель 27, 2023

Fluctuations in brain and behavioral state are supported by broadly projecting neuromodulatory systems. In this study, we use mesoscale two-photon calcium imaging to examine spontaneous activity of cholinergic noradrenergic axons awake mice order determine the interaction between arousal/movement transitions across dorsal cortex at distances separated up 4 mm. We confirm that GCaMP6s within axonal projections both basal forebrain locus coeruleus neurons track arousal, indexed as pupil diameter, changes engagement, reflected bouts whisker movement and/or locomotion. The broad coordination even distant segments indicates these systems can communicate, part, through a global signal, especially relation state. addition coordinated activity, also find evidence subpopulation may exhibit heterogeneity appears be independent our measures By monitoring interneurons cortex, found cells state-dependent (arousal/movement) activity. These results demonstrate provide prominent synchronized signal related state, therefore contribute cortical excitability.

Язык: Английский

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Somatostatin interneurons control the timing of developmental desynchronization in cortical networks

Neuron, Год журнала: 2024, Номер 112(12), С. 2015 - 2030.e5

Опубликована: Апрель 9, 2024

Synchronous neuronal activity is a hallmark of the developing brain. In mouse cerebral cortex, decorrelates during second week postnatal development, progressively acquiring characteristic sparse pattern underlying integration sensory information. The maturation inhibition seems critical for this process, but interneurons involved in crucial transition network cortex remain unknown. Using vivo longitudinal two-photon calcium imaging period that precedes change from highly synchronous to decorrelated activity, we identify somatostatin-expressing (SST+) as modulators switch mice. Modulation SST+ cells accelerates or delays decorrelation cortical process involves regulating parvalbumin-expressing (PV+) interneurons. critically link inputs with local circuits, controlling neural dynamics while modulating other into nascent circuits.

Язык: Английский

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Inhibitory specificity from a connectomic census of mouse visual cortex

Nature, Год журнала: 2025, Номер 640(8058), С. 448 - 458

Опубликована: Апрель 9, 2025

Mammalian cortex features a vast diversity of neuronal cell types, each with characteristic anatomical, molecular and functional properties1. Synaptic connectivity shapes how type participates in the cortical circuit, but mapping rules at resolution distinct types remains difficult. Here we used millimetre-scale volumetric electron microscopy2 to investigate all inhibitory neurons across densely segmented population 1,352 cells spanning layers mouse visual cortex, producing wiring diagram inhibition more than 70,000 synapses. Inspired by classical neuroanatomy, classified based on targeting dendritic compartments developed an excitatory neuron classification reconstructions whole-cell maps synaptic input. Single-cell showed class disinhibitory specialist that targets basket cells. Analysis onto found widespread specificity, many interneurons exhibiting differential spatially intermingled subpopulations. Inhibitory was organized into 'motif groups', diverse sets collectively target both perisomatic same targets. Collectively, our analysis identified new organizing principles for will serve as foundation linking contemporary multimodal atlases diagram.

Язык: Английский

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