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Wiegert, J. S.

Publications and source records attributed to Wiegert, J. S..

At least 19 recordsLinked to original sources

Interplay of pupil-linked arousal and cortical decision computations in mice

Influential frameworks hold that brainstem neuromodulatory systems are rapidly recruited in a top-down manner during decision-making, adapting brain-wide activity and cognitive behavior in real time. Here, we examined the relationship between decision-related frontal cortical activity and pupil responses (a peripheral proxy of neuromodulatory activity) in mice accumulating fluctuating auditory evidence to choose among two options. We found two components of preparatory activity during decision formation, prior to motor action: a non-selective component shared between hemispheres and a choice-selective component that reflected decision uncertainty. Both components were mirrored in pupil responses during the task, which, in turn, scaled with decision uncertainty during decision formation and with uncertainty-modulated prediction errors after experiencing the outcome. Pupil responses also predicted an adjustment of choice behavior at the next trial. Our results point to a recurrent interplay between cortical decision computation and pupil-linked neuromodulation. HIGHLIGHTSO_LIMice switch between evidence integration and heuristic strategies during auditory-informed decisions C_LIO_LIPupil responses track decision uncertainty and uncertainty-modulated prediction errors C_LIO_LIDecision-related frontal cortical activity is coupled with pupil responses C_LIO_LIPupil responses predict an adjustment of future choice behavior C_LI IN SHORTMaheu et al. show that the phasic transitions in the arousal state mirror distinct components of cortical activity, including one directly involved in decision computations. In turn, these pupil-linked arousal responses predicted the strategy adopted at the next trial.

neuroscience↗

Serotonergic and noradrenergic interactions in pupil-linked arousal

Changes in central arousal state shape cortical computations underlying perception, thought, and action. Variations in arousal are accompanied by fluctuations in pupil size. In turn, pupil dynamics are often used as a marker of noradrenaline release from neurons of the locus coeruleus. The serotonergic system of the dorsal raphe also contributes to the brainstem control of waking states. However, little is known about its relationship to noradrenergic activity and arousal dynamics in awake animals. Here, we simultaneously accessed both systems in awake mice and unraveled their unique and joint contributions to pupil-linked arousal. Serotonergic and noradrenergic systems co-fluctuated, and serotonergic dorsal raphe neurons affected pupil size partly via noradrenergic populations in the locus coeruleus. Yet, part of the serotonergic control of pupil dynamics was independent of the locus coeruleus. Our findings challenge common assumptions about the neuromodulatory control of pupil dynamics and illuminate the interplay between distinct neurochemical systems within the arousal network of the brainstem.

neuroscience↗

Functional Connectivity Shapes Spine Stability in the Hippocampus

Synaptic connections between neurons determine the flow of information in the brain. Changes in synaptic weight, along with synapse formation and pruning, reshape the functional connectivity of neural circuits--key mechanisms underlying learning and memory. However, the relationship between functional strength and the structural dynamics of individual glutamatergic synapses in the living mammalian brain remains poorly understood. Specifically, how spine morphology and stability relate to functional adaptations is unclear. Here, we repeatedly recorded excitatory postsynaptic calcium transients in single postsynaptic spines of CA1 neurons in response to optogenetic stimulation of presynaptic CA3 cells in awake, head-fixed mice for over 2 weeks. We found that functional connectivity predicted both the structural stability and spatial proximity of synaptic inputs. Spines with large responses exhibited larger volume and higher stability compared to non-responding spines. Over time, responses were highly variable at individual synapses, but stable at the dendritic level, suggesting that dendritic branches receive stable input despite large fluctuations at individual synapses.

neuroscience↗

Schaffer commissural inputs regulate the stability of CA1 spatial representation

Neuronal representations of the world are dynamic. A striking example is the rapid remapping of the hippocampal spatial code, which occurs even when the environment and behavior remain unchanged. CA3 input to CA1 has been shown to exert a key role in triggering synaptic plasticity in place-encoding CA1 cells -- a phenomenon which could provide the cellular foundations for the remapping of the hippocampal code for space. However, how CA3 input directly contributes to place field formation and remapping of the place code in CA1 remains incompletely understood. By combining longitudinal two-photon calcium imaging of CA1 place cells with optogenetic stimulation of presynaptic CA3 neurons in mice running on a linear treadmill, we demonstrate that CA3 transiently modulates the code for allocentric space in CA1. Activation of CA3 cells both induced a small pool of new CA1 place cells and altered the pre-existing CA1 place code. The latter occurred through an unbiased shift of existing place fields specifically, and not through modifications of place field precision or a loss of place tuning in pre-existing place cells. All CA3-driven changes in the CA1 place code were transient and almost completely disappeared by the next day. Taken together, stimulation of CA3 input to CA1 cells alters CA1-encoded spatial representations, leading to a transient decrease in place field precision rather than an overrepresentation of the stimulation zone. HIGHLIGHTSO_LIOptogenetic CA3 stimulation with two-photon CA1 imaging in behaving mice C_LIO_LIStimulation of CA3 input induces a small pool of new CA1 place cells C_LIO_LIPre-existing CA1 place fields shift without loss of tuning or precision C_LIO_LICA3-driven remapping is unbiased in space and largely disappears by next day C_LI

neuroscience↗

Impulse initiation in engrafted pluripotent stem cell-derived cardiomyocytes can stimulate the recipient heart.

Transplantation of pluripotent stem cell-derived cardiomyocytes is a novel promising cell-based therapeutic approach for patients with heart failure. However, engraftment arrhythmias are a predictable life-threatening complication and represent a major hurdle for clinical translation. Catheter-based electrophysiological analysis suggested that the ventricular arrhythmias were caused by an automaticity of the transplanted cells, but whether impulse generation by transplanted cardiomyocytes can propagate to the host myocardium and override the recipient rhythm has not been directly assessed experimentally. We used optogenetics to specifically activate engrafted cardiomyocytes, which resulted in impulse generation in the engrafted cardiomyocytes and stimulated the recipient heart (4/9 hearts). Thus, our study shows that transplanted cardiomyocytes can electrically couple to the host myocardium and stimulate the recipient heart, providing experimental evidence that cardiomyocyte automaticity can serve as a trigger for ventricular arrhythmias.

pharmacology and toxicology↗

A bistable inhibitory OptoGPCR for multiplexed optogenetic control of neural circuits

Information is transmitted between brain regions through the release of neurotransmitters from long-range projecting axons. Understanding how the activity of such long-range connections contributes to behavior requires efficient methods for reversibly manipulating their function. Chemogenetic and optogenetic tools, acting through endogenous G-protein coupled receptor (GPCRs) pathways, can be used to modulate synaptic transmission, but existing tools are limited in sensitivity, spatiotemporal precision, or spectral multiplexing capabilities. Here we systematically evaluated multiple bistable opsins for optogenetic applications and found that the Platynereis dumerilii ciliary opsin (PdCO) is an efficient, versatile, light-activated bistable GPCR that can suppress synaptic transmission in mammalian neurons with high temporal precision in-vivo. PdCO has superior biophysical properties that enable spectral multiplexing with other optogenetic actuators and reporters. We demonstrate that PdCO can be used to conduct reversible loss-of-function experiments in long-range projections of behaving animals, thereby enabling detailed synapse-specific functional circuit mapping.

neuroscience↗

Dissociating instructive from permissive roles of brain circuits with reversible neural activity manipulations

Recent work has demonstrated that both permanent lesions and acute inactivation experiments can lead to erroneous conclusions about the causal role of brain areas in specific behaviors, casting serious doubt on major avenues by which neuroscientists study the brain. To overcome this challenge, we developed a three-stage optogenetic approach which leverages the ability to precisely control the temporal period of regional inactivation with either brief or sustained illumination, enabling investigators to dissociate between putative permissive and instructive roles of brain areas in behavior. We applied this approach to the mouse primary visual cortex (V1) to probe whether V1 is permissive or instructive for the detection low contrast stimuli. Acute inactivation of V1 drastically suppressed performance, but during persistent inactivation, the animals contrast detection recovered to pre-silencing levels. This recovery was itself reversible, as returning the animals to intermittent V1 inactivation reinstated the behavioral deficit. These results argue that V1 is the default circuit mice use to detect visual stimuli, but in its absence, other regions can compensate for it. This novel, temporally controllable optogenetic perturbation paradigm should be useful in other brain circuits to assess whether they are instructive or permissive in a brain function or behavior.

neuroscience↗

All-optical closed-loop voltage clamp for precise control of muscles and neurons in live animals

Excitable cells can be stimulated or inhibited by optogenetics. Since optogenetic actuation regimes are often static, neurons and circuits can quickly adapt, allowing perturbation, but not true control. Hence, we established an optogenetic voltage-clamp (OVC). The voltage-indicator QuasAr2 provides information for fast, closed-loop optical feedback to the bidirectional optogenetic actuator BiPOLES. Voltage-dependent fluorescence is held within tight margins, thus clamping the cell to distinct potentials. We established the OVC in muscles and neurons of Caenorhabditis elegans, and transferred it to rat hippocampal neurons in slice culture. Fluorescence signals were calibrated to electrically measured potentials, and wavelengths to currents, enabling to determine optical I/V-relationships. The OVC reports on homeostatically altered cellular physiology in mutants and on Ca2+-channel properties, and can dynamically clamp spiking. Combining non-invasive imaging with control capabilities of electrophysiology, the OVC facilitates high-throughput, contact-less electrophysiology in individual cells and paves the way for true optogenetic control in behaving animals.

neuroscience↗

A flexible and versatile system for multicolor fiber photometry and optogenetic manipulation

Fiber photometry is a technique of growing popularity in neuroscientific research. It is widely used to infer brain activity by recording calcium dynamics in genetically defined populations of neurons. Aside from the wide variety of calcium indicators, other genetically encoded biosensors have recently been engineered to measure membrane potential, neurotransmitter release, pH, or various cellular metabolites, such as ATP or cAMP. Due to the spectral characteristics of these molecular tools, different assemblies of optical hardware are usually needed to reveal the full potential of different biosensors. In addition, the combination of multiple biosensors in one experiment often requires the investment in more complex equipment, which limits the flexibility of the experimental design. Such constraints often hamper a straightforward implementation of new molecular tools, evaluation of their performance in vivo, and design of new experimental paradigms - especially if the financial budget is a limiting factor. Here, we propose a novel approach for fiber photometry recordings, based on a multimode optical fused-fiber coupler (FFC) for both light delivery and collection. Recordings can readily be combined with optogenetic manipulations in a single device without the requirement for dichroic beam-splitters. In combination with a multi-color light source and appropriate emission filters, our approach offers remarkable flexibility in experimental design and facilitates the implication of new molecular tools in vivo at minimal cost. The ease of assembly, operation, characterization, and customization of this platform holds the potential to foster the development of experimental strategies for multicolor fused fiber photometry (FFP) combined with optogenetics far beyond its current state. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=186 SRC="FIGDIR/small/484590v1_ufig1.gif" ALT="Figure 1"> View larger version (45K): org.highwire.dtl.DTLVardef@13abbdcorg.highwire.dtl.DTLVardef@10384d7org.highwire.dtl.DTLVardef@67938org.highwire.dtl.DTLVardef@1005aee_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LIFused Fiber Photometry (FFP) enables straightforward monitoring and manipulation of brain activity C_LIO_LIFFP allows monitoring of indicators with virtually any spectral characteristics C_LIO_LIFFP is compatible with simultaneous optogenetic manipulation C_LIO_LIEasy assembly, characterization, and customization C_LI

neuroscience↗

Temporally extended and reversible neuronal silencing with Aion

Optogenetic silencing allows to reveal the necessity of selected neuronal populations for various neurophysiological functions ranging from specific behaviors to neuronal network activity. An ideal optogenetic silencing tool should be switchable between active and inactive states with precise timing while preserving its activity in the absence of light until switched to an inactive state. Although bistable anion-conducting channelrhodopsins (ACRs) were previously engineered to reach this goal, their conducting state life time was limited to only a few minutes and some ACRs were not fully switchable. Here we report Aion, a bistable ACR displaying a long-lasting open state with a spontaneous closing time constant close to 15 min. Moreover, Aion can be switched between the open and closed state with millisecond precision using blue and orange light, respectively. The long conducting state enables overnight silencing of neurons with minimal light exposure. We further generated trafficking-optimized versions of Aion, which show enhanced membrane localization and allow precisely timed, long-lasting all-optical control of nociceptive responses in larvae of Drosophila melanogaster.

neuroscience↗

Targeting Noradrenergic Neurons of the Locus Coeruleus: A Comparison of Model Systems and Strategies

The locus coeruleus (LC) norepinephrine (NE) system is involved in a variety of physiological and pathophysiological processes. Refining our understanding of LC function largely relies on selective transgene expression in LC-NE neurons, allowing targeted manipulation and readout of noradrenergic neurons. Here, we performed a side-by-side comparison of the most commonly used strategies to genetically target the LC, including different cre driver lines and promoter-mediated transgene expression. We report differences between these strategies in terms of transgene expression efficacy and specificity. Parallelly, we found no behavioral alterations in cre-expressing mice of any mouse line compared to wild-type littermates. Finally, to further facilitate the investigation of LC-NE function, we created a suite of constructs, including a reporter protein, a calcium indicator, and a light-driven cation channel, whose expression is mediated by the previously described PRSx8 promoter. These constructs allow identification, monitoring, and manipulation of LC-NE activity either in wild-type mice, or in combination with tissue-specific manipulations of different cre driver lines. The results of our study are crucial for the interpretation of previous experiments using the respective targeting strategies, as well as for the design of future studies. BlurbThis study reports substantial heterogeneity in transgene expression patterns when using different strategies to virally transduce noradrenergic neurons of the locus coeruleus, highlighting the need to carefully validate each experimental approach in order to avoid misinterpretations when studying locus coeruleus function. Highlights- Heterogeneous expression patterns with different targeting strategies for LC-NE neurons - Substantial ectopic transgene expression observed in some approaches - No behavioral alterations in heterozygous cre mice as compared to wild-types - PRSx8-driven reporters, indicators and actuators as a resource to investigate LC function

neuroscience↗

Contractile force of transplanted cardiomyocytes contributes to heart function after injury

Transplantation of pluripotent stem cell-derived cardiomyocytes represents an innovative therapeutic strategy for heart failure. Studies in small and large animals have demonstrated functional recovery of left ventricular function after cardiomyocyte transplantation1-4, and first clinical studies are currently underway5. Yet, the mechanism of action underlying graft-induced benefit is unknown6. Here we demonstrate that transplanted cardiomyocytes actively contribute to heart function. We transplanted cardiomyocytes with an optogenetic off-on switch in a guinea pig cardiac injury model. Light-induced inhibition of engrafted cardiomyocyte contractility resulted in a rapid decrease of left ventricular function that was fully reversible with the offset of photostimulation. Hence, our optogenetic approach demonstrated that transplanted cardiomyocytes actively participate in heart function, supporting the hypothesis that the delivery of new force-generating myocardium can serve as a regenerative therapeutic strategy.

pharmacology and toxicology↗

Improved two-photon imaging of GPCR-based optogenetic neurotransmitter sensors using orthogonally polarized excitation

Fluorescent proteins such as GFP are best excited by light that is polarized parallel to the dipole axis of the fluorophore. In most cases, fluorescent proteins are randomly oriented, resulting in unbiased images even when polarized light is used for excitation, e.g. in two-photon microcopy. Here we reveal a surprisingly strong polarization sensitivity in a class of GPCR-based neurotransmitter sensors where the fluorophore is anchored on both ends. In tubular structures such as dendrites, this effect led to a complete loss of membrane signal in dendrites running parallel to the polarization direction of the excitation beam. Our data reveal a major problem for two-photon measurements of neurotransmitter concentration that has not been recognized by the neuroscience community. To remedy the sensitivity to dendritic orientation, we designed an optical device that generates interleaved pulse trains of orthogonal polarization, removing the orientation bias from images. The passive device, which we inserted in the beam path of an existing two-photon microscope, also removed the strong direction bias in second harmonic generation (SHG) images. We conclude that for optical measurements of transmitter concentration with GPCR-based sensors, orthogonally polarized excitation is essential.

neuroscience↗

Optogenetic silencing of neurotransmitter release with a naturally occurring invertebrate rhodopsin

Information is carried between brain regions through neurotransmitter release from axonal presynaptic terminals. Understanding the functional roles of defined neuronal projection pathways in cognitive and behavioral processes requires temporally precise manipulation of their activity in vivo. However, existing optogenetic tools have low efficacy and off-target effects when applied to presynaptic terminals, while chemogenetic tools are difficult to control in space and time. Here, we show that a targeting-enhanced mosquito homologue of the vertebrate encephalopsin (eOPN3) can effectively suppress synaptic transmission through the Gi/o signaling pathway. Brief illumination of presynaptic terminals expressing eOPN3 triggers a lasting suppression of synaptic output that recovers spontaneously within minutes in vitro as well as in vivo. In freely moving mice, eOPN3-mediated suppression of dopaminergic nigrostriatal afferents leads to an ipsiversive rotational bias. We conclude that eOPN3 can be used to selectively suppress neurotransmitter release at synaptic terminals with high spatiotemporal precision, opening new avenues for functional interrogation of long-range neuronal circuits in vivo.

neuroscience↗

The vesicular release probability sets the strength of individual Schaffer collateral synapses

Information processing in the brain is controlled by quantal release of neurotransmitters, a tightly regulated process. From ultrastructural analysis, it is known that presynaptic boutons along single axons differ in the number of vesicles docked at the active zone. It is not clear whether the probability of these vesicles to get released (pves) is homogenous or also varies between individual boutons. Here, we optically measure evoked transmitter release at individual Schaffer collateral synapses at different calcium concentrations, using the genetically encoded glutamate sensor iGluSnFR. Fitting a binomial model to measured response amplitude distributions allowed us to extract the quantal parameters N, pves, and q. We find that Schaffer collateral boutons typically release single vesicles under low pves conditions and switch to multivesicular release in high calcium saline. Analyzing the variability of quantal parameters, we conclude that the vesicular release probability rather than the number of readily releasable vesicles or their transmitter content determines the potency of individual boutons.

neuroscience↗

BiPOLES: a tool for bidirectional dual-color optogenetic control of neurons

Optogenetic manipulation of neuronal activity has become an indispensable experimental strategy in neuroscience research. A large repertoire of excitatory and inhibitory tools allows precise activation or inhibition of genetically targetable neuronal populations. However, an optogenetic tool for reliable bidirectional control of neuronal activity allowing both up- and downregulation of the same neurons in a single experiment is still missing. Here we report BiPOLES, an optogenetic tool for potent excitation and inhibition of the same population of neurons with light of two different colors. BiPOLES consists of an inhibitory, blue-light-sensitive anion-conducting channelrhodopsin fused to an excitatory, red-light-sensitive cation-conducting channelrhodopsin in a single, trafficking-optimized tandem protein. BiPOLES enables multiple new applications including potent dual-color spiking and silencing of the same neurons in vivo and dual-color optogenetic control of two independent neuronal populations.

neuroscience↗

Anesthetics uniquely decorrelate hippocampal network activity, alter spine dynamics and affect memory consolidation

General anesthesia is characterized by reversible loss of consciousness accompanied by transient amnesia. Yet, long-term memory impairment is an undesirable side-effect. How different types of general anesthetics (GAs) affect the hippocampus, a brain region central to memory formation and consolidation, is poorly understood. Using extracellular recordings, chronic 2-photon imaging and behavioral analysis, we monitor the effects of isoflurane (Iso), medetomidine/midazolam/fentanyl (MMF), and ketamine/xylazine (Keta/Xyl) on network activity and structural spine dynamics in the hippocampal CA1 area of adult mice. GAs robustly reduced spiking activity, decorrelated cellular ensembles, albeit with distinct activity signatures, and altered spine dynamics. CA1 network activity under all three anesthetics was different to natural sleep. Iso anesthesia most closely resembled unperturbed activity during wakefulness and sleep, and network alterations recovered more readily than with Keta/Xyl and MMF. Correspondingly, memory consolidation was impaired after exposure to Keta/Xyl and MMF, but not Iso. Thus, different anesthetics distinctly alter hippocampal network dynamics, synaptic connectivity, and memory consolidation, with implications for GA strategy appraisal in animal research and clinical settings.

neuroscience↗

Long vs short-term synaptic learning rules after optogenetic spike-timing-dependent plasticity

Spike-timing-dependent plasticity (STDP) is a candidate mechanism for information storage in the brain, but the whole-cell recordings required for the experimental induction of STDP are typically limited to one hour. This mismatch of time scales is a long-standing weakness in synaptic theories of memory. Here we use spectrally separated optogenetic stimulation to fire precisely timed action potentials (spikes) in CA3 and CA1 pyramidal cells. Twenty minutes after optogenetic induction of STDP (oSTDP), we observed timing-dependent depression (tLTD) and timing-dependent potentiation (tLTP), depending on the sequence of spiking. As oSTDP does not require electrodes, we could also assess the strength of these paired connections three days later. At this late time point, late tLTP was observed for both causal (CA3 before CA1) and anti-causal (CA1 before CA3) timing, but not for asynchronous activity patterns ({Delta}t = 50 ms). Blocking activity after induction of oSTDP prevented stable potentiation. Our results confirm that neurons wire together if they fire together, but suggest that synaptic depression after anti-causal activation (tLTD) is a transient phenomenon. HighlightsO_LIOptogenetic induction of spike-timing-dependent plasticity at Schaffer collateral synapses C_LIO_LICausal pairing induces potentiation whereas anti-causal pairing induces depression during patch-clamp recordings. C_LIO_LIThree days after optogenetic induction, the consequence of STDP is potentiation (tLTP) irrespective of spiking order. C_LIO_LILate tLTP requires ongoing activity in the days following oSTDP. C_LI Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=138 SRC="FIGDIR/small/863365v4_ufig1.gif" ALT="Figure 1"> View larger version (19K): org.highwire.dtl.DTLVardef@1937ad5org.highwire.dtl.DTLVardef@16566f8org.highwire.dtl.DTLVardef@4b4a73org.highwire.dtl.DTLVardef@104fc97_HPS_FORMAT_FIGEXP M_FIG C_FIG

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