In Vivo Optogenetic Stimulation of Neocortical Excitatory Neurons Drives Brain-State-Dependent Inhibition

Abstract Introduction The neural circuits controlling rapid eye movement (REM) sleep, and in particular the role of the medulla in regulating this brain state, remains an active area of study. Previous electrophysiological recordings in the dorsomedial medulla (DM) and electrical stimulation experiments suggested an important role of this area in the control of REM sleep. However the identity of the involved neurons and their precise role in REM sleep regulation are still unclear. Methods The properties of DM GAD2 neurons in mice were investigated through stereotaxic injection of CRE-dependent viruses in conjunction with implantation of electrodes for electroencephalogram (EEG) and electromyogram (EMG) recordings and optic fibers. Experiments included in vivo calcium imaging (fiber photometry) across sleep and wake states, optogenetic stimulation of cell bodies, chemogenetic excitation and suppression (DREADDs), and connectivity mapping using viral tracing and optogenetics. Results Imaging the calcium activity of DM GAD2 neurons in vivo indicates that these neurons are most active during REM sleep. Optogenetic stimulation of DM GAD2 neurons reliably triggered transitions into REM sleep from NREM sleep. Consistent with this, chemogenetic activation of DM GAD2 neurons increased the amount of REM sleep while inhibition suppressed its occurrence and enhanced NREM sleep. Anatomical tracing revealed that DM GAD2 neurons project to several areas involved in sleep / wake regulation including the wake-promoting locus coeruleus (LC) and the REM sleep-suppressing ventrolateral periaquaductal gray (vlPAG). Optogenetic activation of axonal projections from DM to LC, and DM to vlPAG was sufficient to induce REM sleep. Conclusion These experiments demonstrate that DM inhibitory neurons expressing GAD2 powerfully promote initiation of REM sleep in mice. These findings further characterize the dorsomedial medulla as a critical structure involved in REM sleep regulation and inform future investigations of the REM sleep circuitry. Support R01 HL149133

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0154 Role of Noradrenergic Projection to the Preoptic Area in Regulation of Arousal

SLEEP ◽

10.1093/sleep/zsaa056.152 ◽

2020 ◽

Vol 43 (Supplement_1) ◽

pp. A61-A61

Author(s):

H Antila ◽

I Kwak ◽

I Covarrubias ◽

J Baik ◽

J Hong ◽

...

Keyword(s):

Preoptic Area ◽

Brain Regions ◽

Brain State ◽

Research Fellowship ◽

Optogenetic Stimulation ◽

Long Time ◽

Brain States ◽

Stimulation Of

Abstract Introduction Locus coeruleus (LC) is a noradrenergic nucleus in the brainstem involved in the regulation of attention, arousal, mood and sensory gating. LC projects to multiple brain regions and recent development of novel systems neuroscience tools allows the dissection of projection-specific LC function in more detail. One of the regions with noradrenergic projection is the preoptic area of the hypothalamus (POA). POA has been shown to contain neurons that are important for regulating sleep, and we have examined the function of the LC projection to the POA in sleep and arousal. Methods We used optogenetics, chemogenetics, fiber photometry and in vivo electrophysiology to study the function of LC noradrenergic projection to the POA. Results Norepinephrine release in the POA fluctuates with brain state changes indicating that the LC to POA projection may be involved in regulating sleep and arousal. Optogenetic stimulation of LC fibers in the POA promotes wakefulness. Furthermore, optogenetic stimulation of the LC fibers in the POA modulates the activity of sleep- and wake-active neurons. Conclusion We have identified the role of the LC noradrenergic projection to the POA in the regulation of brain states. Stimulation of the LC fibers in the POA promotes wakefulness and modulates the activity dynamics of sleep- and wake-active neurons in the POA. Our results provide more detailed information about the role of this specific projection, which has been known to exist for a long time, but with insufficient in vivo evidence of its precise function. Support Sigrid Juselius foundation, Alfred P. Sloan Research Fellowship in Neuroscience, The Whitehall foundation grant, McCabe Fund Award, NARSAD Young Investigator Award.

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Differential cell-type dependent brain state modulations of sensory representations in the non-lemniscal mouse inferior colliculus

Communications Biology ◽

10.1038/s42003-019-0602-4 ◽

2019 ◽

Vol 2 (1) ◽

Cited By ~ 1

Author(s):

Chenggang Chen ◽

Sen Song

Keyword(s):

Inferior Colliculus ◽

Calcium Imaging ◽

Inhibitory Neurons ◽

Brain State ◽

Spectral Tuning ◽

State Dependent ◽

Excitatory Neurons ◽

Sensory Responses ◽

Highly Correlated

Abstract Sensory responses of the neocortex are strongly influenced by brain state changes. However, it remains unclear whether and how the sensory responses of the midbrain are affected. Here we addressed this issue by using in vivo two-photon calcium imaging to monitor the spontaneous and sound-evoked activities in the mouse inferior colliculus (IC). We developed a method enabling us to image the first layer of non-lemniscal IC (IC shell L1) in awake behaving mice. Compared with the awake state, spectral tuning selectivity of excitatory neurons was decreased during isoflurane anesthesia. Calcium imaging in behaving animals revealed that activities of inhibitory neurons were highly correlated with locomotion. Compared with stationary periods, spectral tuning selectivity of excitatory neurons was increased during locomotion. Taken together, our studies reveal that neuronal activities in the IC shell L1 are brain state dependent, whereas the brain state modulates the excitatory and inhibitory neurons differentially.

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Opposing Influence of Sensory and Motor Cortical Input on Striatal Circuitry and Choice Behavior

10.1101/446708 ◽

2018 ◽

Author(s):

Christian R. Lee ◽

Alex J. Yonk ◽

Joost Wiskerke ◽

Kenneth G. Paradiso ◽

James M. Tepper ◽

...

Keyword(s):

Ex Vivo ◽

Dorsal Striatum ◽

Behavioral Effect ◽

Projection Neurons ◽

Cortical Input ◽

Optogenetic Stimulation ◽

Main Input ◽

Opposing Effects ◽

Stimulation Of

SummaryThe striatum is the main input nucleus of the basal ganglia and is a key site of sensorimotor integration. While the striatum receives extensive excitatory afferents from the cerebral cortex, the influence of different cortical areas on striatal circuitry and behavior is unknown. Here we find that corticostriatal inputs from whisker-related primary somatosensory (S1) and motor (M1) cortex differentially innervate projection neurons and interneurons in the dorsal striatum, and exert opposing effects on sensory-guided behavior. Optogenetic stimulation of S1-corticostriatal afferents in ex vivo recordings produced larger postsynaptic potentials in striatal parvalbumin (PV)-expressing interneurons than D1- or D2-expressing spiny projection neurons (SPNs), an effect not observed for M1-corticostriatal afferents. Critically, in vivo optogenetic stimulation of S1-corticostriatal afferents produced task-specific behavioral inhibition, which was bidirectionally modulated by striatal PV interneurons. Optogenetic stimulation of M1 afferents produced the opposite behavioral effect. Thus, our results suggest opposing roles for sensory and motor cortex in behavioral choice via distinct influences on striatal circuitry.

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Parylene photonics: a flexible, broadband optical waveguide platform with integrated micromirrors for biointerfaces

Microsystems & Nanoengineering ◽

10.1038/s41378-020-00186-2 ◽

2020 ◽

Vol 6 (1) ◽

Cited By ~ 1

Author(s):

Jay W. Reddy ◽

Maya Lassiter ◽

Maysamreza Chamanzar

Keyword(s):

Optical Waveguides ◽

Brain Activity ◽

Low Loss ◽

Parylene C ◽

Optogenetic Stimulation ◽

Patterned Illumination ◽

The Brain ◽

532 Nm ◽

Stimulation Of

Abstract Targeted light delivery into biological tissue is needed in applications such as optogenetic stimulation of the brain and in vivo functional or structural imaging of tissue. These applications require very compact, soft, and flexible implants that minimize damage to the tissue. Here, we demonstrate a novel implantable photonic platform based on a high-density, flexible array of ultracompact (30 μm × 5 μm), low-loss (3.2 dB/cm at λ = 680 nm, 4.1 dB/cm at λ = 633 nm, 4.9 dB/cm at λ = 532 nm, 6.1 dB/cm at λ = 450 nm) optical waveguides composed of biocompatible polymers Parylene C and polydimethylsiloxane (PDMS). This photonic platform features unique embedded input/output micromirrors that redirect light from the waveguides perpendicularly to the surface of the array for localized, patterned illumination in tissue. This architecture enables the design of a fully flexible, compact integrated photonic system for applications such as in vivo chronic optogenetic stimulation of brain activity.

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Optogenetic Stimulation of Adrenergic C1 Neurons Causes Sleep State–Dependent Cardiorespiratory Stimulation and Arousal with Sighs in Rats

American Journal of Respiratory and Critical Care Medicine ◽

10.1164/rccm.201407-1262oc ◽

2014 ◽

Vol 190 (11) ◽

pp. 1301-1310 ◽

Cited By ~ 48

Author(s):

Peter G. R. Burke ◽

B. Stephen ◽

Melissa B. Coates ◽

Kenneth E. Viar ◽

Ruth L. Stornetta ◽

...

Keyword(s):

Sleep State ◽

Optogenetic Stimulation ◽

State Dependent ◽

Stimulation Of

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Network state-dependent inhibition of identified hippocampal CA3 axo-axonic cells in vivo

Nature Neuroscience ◽

10.1038/nn.3550 ◽

2013 ◽

Vol 16 (12) ◽

pp. 1802-1811 ◽

Cited By ~ 79

Author(s):

Tim J Viney ◽

Balint Lasztoczi ◽

Linda Katona ◽

Michael G Crump ◽

John J Tukker ◽

...

Keyword(s):

Network State ◽

State Dependent ◽

Dependent Inhibition ◽

Hippocampal Ca3

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Optogenetic activation of muscle contraction in vivo

10.1101/2020.07.07.192377 ◽

2020 ◽

Author(s):

Elahe Ganji ◽

C. Savio Chan ◽

Christopher W. Ward ◽

Megan L. Killian

Keyword(s):

Skeletal Muscle ◽

Electrical Stimulation ◽

Muscle Contraction ◽

Fluorescent Imaging ◽

Triceps Surae ◽

Light Activation ◽

Non Invasive ◽

Optogenetic Stimulation ◽

Stimulation Of

AbstractOptogenetics is an emerging alternative to traditional electrical stimulation to initiate action potentials in activatable cells both ex vivo and in vivo. Optogenetics has been commonly used in mammalian neurons and more recently, it has been adapted for activation of cardiomyocytes and skeletal muscle. Therefore, the aim of this study was to evaluate the stimulation feasibility and sustain isometric muscle contraction and limit decay for an extended period of time (1s), using non-invasive transdermal light activation of skeletal muscle (triceps surae) in vivo. We used inducible Cre recombination to target expression of Channelrhodopsin-2 (ChR2(H134R)-EYFP) in skeletal muscle (Acta1-Cre) in mice. Fluorescent imaging confirmed that ChR2 expression is localized in skeletal muscle and does not have specific expression in sciatic nerve branch, therefore, allowing for non-nerve mediated optical stimulation of skeletal muscle. We induced muscle contraction using transdermal exposure to blue light and selected 10Hz stimulation after controlled optimization experiments to sustain prolonged muscle contraction. Increasing the stimulation frequency from 10Hz to 40Hz increased the muscle contraction decay during prolonged 1s stimulation, highlighting frequency dependency and importance of membrane repolarization for effective light activation. Finally, we showed that optimized pulsed optogenetic stimulation of 10 Hz resulted in comparable ankle torque and contractile functionality to that of electrical stimulation. Our results demonstrate the feasibility and repeatability of non-invasive optogenetic stimulation of muscle in vivo and highlight optogenetic stimulation as a powerful tool for non-invasive in vivo direct activation of skeletal muscle.

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All-optical electrophysiology reveals brain-state dependent changes in hippocampal subthreshold dynamics and excitability

10.1101/281618 ◽

2018 ◽

Cited By ~ 16

Author(s):

Yoav Adam ◽

Jeong J. Kim ◽

Shan Lou ◽

Yongxin Zhao ◽

Daan Brinks ◽

...

Keyword(s):

High Speed ◽

Near Infrared ◽

Neuronal Excitability ◽

Signal To Noise Ratio ◽

Brain State ◽

Promising Tool ◽

State Dependent ◽

Subthreshold Voltage ◽

Targeted Gene Expression

AbstractA technology to record membrane potential from multiple neurons, simultaneously, in behaving animals will have a transformative impact on neuroscience research1. Parallel recordings could reveal the subthreshold potentials and intercellular correlations that underlie network behavior2. Paired stimulation and recording can further reveal the input-output properties of individual cells or networks in the context of different brain states3. Genetically encoded voltage indicators are a promising tool for these purposes, but were so far limited to single-cell recordings with marginal signal to noise ratio (SNR) in vivo4-6. We developed improved near infrared voltage indicators, high speed microscopes and targeted gene expression schemes which enabled recordings of supra- and subthreshold voltage dynamics from multiple neurons simultaneously in mouse hippocampus, in vivo. The reporters revealed sub-cellular details of back-propagating action potentials, correlations in sub-threshold voltage between multiple cells, and changes in dynamics associated with transitions from resting to locomotion. In combination with optogenetic stimulation, the reporters revealed brain state-dependent changes in neuronal excitability, reflecting the interplay of excitatory and inhibitory synaptic inputs. These tools open the possibility for detailed explorations of network dynamics in the context of behavior.

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Optogenetic stimulation of Dbx1 neurons enhances the respiratory‐sympathetic coupling in in vivo CIH mice

The FASEB Journal ◽

10.1096/fasebj.2021.35.s1.04446 ◽

2021 ◽

Vol 35 (S1) ◽

Author(s):

Marlusa Karlen‐Amarante ◽

Alyssa Huff ◽

Nathan Baertsch ◽

Debora Colombari ◽

Jan‐Marino Ramirez

Keyword(s):

Optogenetic Stimulation ◽

Stimulation Of

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