A genetically encoded sensor for in vivo imaging of orexin neuropeptides

Orexins (also called hypocretins) are hypothalamic neuropeptides that carry out essential functions in the central nervous system; however, little is known about their release and range of action in vivo owing to the limited resolution of current detection technologies. Here we developed a genetically encoded orexin sensor (OxLight1) based on the engineering of circularly permutated green fluorescent protein into the human type-2 orexin receptor. In mice OxLight1 detects optogenetically evoked release of endogenous orexins in vivo with high sensitivity. Photometry recordings of OxLight1 in mice show rapid orexin release associated with spontaneous running behavior, acute stress and sleep-to-wake transitions in different brain areas. Moreover, two-photon imaging of OxLight1 reveals orexin release in layer 2/3 of the mouse somatosensory cortex during emergence from anesthesia. Thus, OxLight1 enables sensitive and direct optical detection of orexin neuropeptides with high spatiotemporal resolution in living animals. OxLight1 is a genetically encoded sensor for the orexin neuropeptides. It has been applied in fiber photometry recordings and two-photon imaging in mice during a variety of behaviors.

• Publication year

  2022

• Venue

  Nature Methods

• Publication date

  2022-02-01

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1038/s41592-021-01390-2PMID 35145320PMCID 8831244
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Structures of active-state orexin receptor 2 rationalize peptide and small-molecule agonist recognition and receptor activation

  2021cited by this paper
• A genetically encoded sensor for measuring serotonin dynamics

  2021cited by this paper
• Next-generation GRAB sensors for monitoring dopaminergic activity in vivo

  2020cited by this paper
• An optimized acetylcholine sensor for monitoring in vivo cholinergic activity

  2020cited by this paper
• Role of spontaneous and sensory orexin network dynamics in rapid locomotion initiation

  2020cited by this paper
• An expanded palette of dopamine sensors for multiplex imaging in vivo

  2020cited by this paper
• Aversive state processing in the posterior insular cortex

  2019cited by this paper
• The catalytic function of the gephyrin‐binding protein IQSEC3 regulates neurotransmitter‐specific matching of pre‐ and post‐synaptic structures in primary hippocampal cultures

  2018cited by this paper
• Ultrafast neuronal imaging of dopamine dynamics with designed genetically encoded sensors

  2018cited by this paper
• Mini G protein probes for active G protein–coupled receptors (GPCRs) in live cells

  2018cited by this paper
• A genetically encoded fluorescent sensor for rapid and specific in vivo detection of norepinephrine

  2018cited by this paper
• The neurobiological basis of narcolepsy

  2018cited by this paper
• PARALLEL CIRCUITS FROM THE BED NUCLEI OF STRIA TERMINALIS TO THE LATERAL HYPOTHALAMUS DRIVE OPPOSING EMOTIONAL STATES

  2018cited by this paper
• Deciphering the molecular mechanism responsible for GCaMP6m's Ca2+-dependent change in fluorescence

  2017cited by this paper
• Projection-Target-Defined Effects of Orexin and Dynorphin on VTA Dopamine Neurons.

  2017cited by this paper
• Mapping the Hypocretin/Orexin Neuronal System: An Unexpectedly Productive Journey

  2017cited by this paper
• Accumbal D2 cells orchestrate innate risk-avoidance according to orexin signals

  2017cited by this paper
• Suite2p: beyond 10,000 neurons with standard two-photon microscopy

  2016cited by this paper
• Awake dynamics and brain-wide direct inputs of hypothalamic MCH and orexin networks

  2016cited by this paper
• Slow Bursting Neurons of Mouse Cortical Layer 6b Are Depolarized by Hypocretin/Orexin and Major Transmitters of Arousal

  2016cited by this paper
• High-accuracy reference standards for two-photon absorption in the 680-1050 nm wavelength range.

  2016cited by this paper
• Sensitive red protein calcium indicators for imaging neural activity

  2016cited by this paper
• Design and performance of an ultra-flexible two-photon microscope for in vivo research.

  2015cited by this paper
• Hypocretin (orexin) facilitates reward by attenuating the antireward effects of its cotransmitter dynorphin in ventral tegmental area

  2014cited by this paper
• Motivational activation: a unifying hypothesis of orexin/hypocretin function

  2014cited by this paper
• Clustal Omega, accurate alignment of very large numbers of sequences.

  2014cited by this paper
• The role of orexin in motivated behaviours

  2014cited by this paper
• Activation of central orexin/hypocretin neurons by dietary amino acids.

  2011cited by this paper
• Circular Polymerase Extension Cloning of Complex Gene Libraries and Pathways

  2009cited by this paper
• A KEY ROLE FOR OREXIN IN PANIC ANXIETY

  2009cited by this paper
• General anaesthesia: from molecular targets to neuronal pathways of sleep and arousal

  2008cited by this paper
• The neural circuit of orexin (hypocretin): maintaining sleep and wakefulness

  2007cited by this paper
• Orexin‐A is composed of a highly conserved C‐terminal and a specific, hydrophilic N‐terminal region, revealing the structural basis of specific recognition by the orexin‐1 receptor

  2006cited by this paper
• Tandem-pore K+ channels mediate inhibition of orexin neurons by glucose.

  2006cited by this paper
• A role for lateral hypothalamic orexin neurons in reward seeking

  2005cited by this paper
• Behavioral correlates of activity in identified hypocretin/orexin neurons.

  2005cited by this paper
• Distinct narcolepsy syndromes in Orexin receptor-2 and Orexin null mice: molecular genetic dissection of Non-REM and REM sleep regulatory processes.

  2003cited by this paper
• Distinct Recognition of OX1 and OX2Receptors by Orexin Peptides

  2003cited by this paper
• Genetic ablation of orexin neurons in mice results in narcolepsy, hypophagia, and obesity.

  2001cited by this paper
• Differential expression of orexin receptors 1 and 2 in the rat brain

  2001cited by this paper
• Hypocretin (orexin) deficiency in human narcolepsy.

  2000cited by this paper
• Reduced Number of Hypocretin Clinical Study Neurons in Human Narcolepsy

  2000cited by this paper
• A mutation in a case of early onset narcolepsy and a generalized absence of hypocretin peptides in human narcoleptic brains

  2000cited by this paper
• Characterization of recombinant human orexin receptor pharmacology in a Chinese hamster ovary cell‐line using FLIPR

  1999cited by this paper
• Hypothalamic Hypocretin (Orexin): Robust Innervation of the Spinal Cord

  1999cited by this paper
• Hypothalamic Hypocretin (Orexin): Robust Innervation of the Spinal Cord

  1999cited by this paper
• The sleep disorder canine narcolepsy is caused by a mutation in the hypocretin (orexin) receptor 2 gene.

  1999cited by this paper
• Solution structure of a new hypothalamic neuropeptide, human hypocretin-2/orexin-B.

  1999cited by this paper
• Narcolepsy in orexin knockout mice: molecular genetics of sleep regulation.

  1999cited by this paper
• Orexins and orexin receptors: a family of hypothalamic neuropeptides and G protein-coupled receptors that regulate feeding behavior.

  1998cited by this paper
• The hypocretins: hypothalamus-specific peptides with neuroexcitatory activity.

  1998cited by this paper
• Neurons Containing Hypocretin (Orexin) Project to Multiple Neuronal Systems

  1998cited by this paper
• Orexins and orexin receptors: a family of hypothalamic neuropeptides and G protein-coupled receptors that regulate feeding behavior.

  1998cited by this paper
• Measurement of two-photon excitation cross sections of molecular fluorophores with data from 690 to 1050 nm

  1996cited by this paper

• Empathy and prosocial behavior powered by orexin-driven theta oscillations.

  2026cites this paper
• Rising Stars: In vivo monitoring of neurochemical dynamics by genetically encoded neuromodulator sensors.

  2026cites this paper
• Next-generation multicolor indicators for in vivo imaging of norepinephrine.

  2026cites this paper
• Current and emerging tools for studying animal models of sleep and autism.

  2025cites this paper
• Development of a genetically encoded melanocortin sensor for high sensitivity intravital imaging

  2025cites this paper
• Development of a genetically encoded melanocortin sensor for high sensitivity in vivo imaging

  2025cites this paper
• Apelin as a CNS-specific pathway for fenestrated capillary formation in the choroid plexus

  2025influential citation
• Spatiotemporal dynamics of CXCL10 encode contextual immune information revealed by the genetically encoded fluorescent sensor.

  2025cites this paper
• In vivo measurement of an Apelin gradient with a genetically encoded APLNR conformation biosensor

  2025cites this paper
• Orexinergic projections to substantia innominata mediate arousal and analgesia.

  2025cites this paper
• [Real-time measurement of neuromodulators using GRAB sensors].

  2025cites this paper
• Mapping and decoding neuropeptide signaling networks in nervous system function

  2025cites this paper
• An integrated microfluidic and fluorescence platform for probing in vivo neuropharmacology.

  2025cites this paper
• Genetically encoded sensors illuminate in vivo detection for neurotransmission: Development, application, and optimization strategies

  2025cites this paper
• Comparative perspectives on neuropeptide function and social isolation.

  2025cites this paper
• Live imaging of paracrine signaling: Advances in visualization and tracking techniques

  2025cites this paper
• A high-performance GRAB sensor reveals differences in the dynamics and molecular regulation between neuropeptide and neurotransmitter release

  2025cites this paper
• Orexin population activity precisely reflects net body movement across behavioral and metabolic states

  2025cites this paper
• Multi-fiber array-based photometry system for multi-regional functional mapping in the mouse brain

  2025cites this paper
• Neuromodulator and neuropeptide sensors and probes for precise circuit interrogation in vivo

  2024cites this paper
• Neuropeptide signaling network of Caenorhabditis elegans: from structure to behavior

  2024cites this paper
• Orexinergic projections to substantia innominata mediate arousal and analgesia

  2024cites this paper
• A light at the end of the axon: genetically encoded fluorescent indicators shine light on the dopamine system

  2024cites this paper
• Imaging different cell populations in the mouse olfactory bulb using the genetically encoded voltage indicator ArcLight

  2024cites this paper
• Molecular Spies in Action: Genetically Encoded Fluorescent Biosensors Light up Cellular Signals

  2024cites this paper
• Engineering, applications, and future perspectives of GPCR‐based genetically encoded fluorescent indicators for neuromodulators

  2024cites this paper
• Probing PAC1 receptor activation across species with an engineered sensor

  2024cites this paper
• A Bright Future? A Perspective on Class C GPCR Based Genetically Encoded Biosensors

  2024cites this paper
• Orexin Neurons to Sublaterodorsal Tegmental Nucleus Pathway Prevents Sleep Onset REM Sleep-Like Behavior by Relieving the REM Sleep Pressure.

  2024cites this paper
• Genetically Encoded Sensors for the In Vivo Detection of Neurochemical Dynamics.

  2024cites this paper
• Orexin neurons track temporal features of blood glucose in behaving mice

  2024cites this paper
• A new GRAB sensor reveals differences in the dynamics and molecular regulation between neuropeptide and neurotransmitter release

  2024cites this paper
• Probing neuronal activity with genetically encoded calcium and voltage fluorescent indicators.

  2024cites this paper
• International Union of Basic and Clinical Pharmacology CXIV: Orexin Receptor Function, Nomenclature and Pharmacology

  2024cites this paper
• Unlocking opioid neuropeptide dynamics with genetically encoded biosensors

  2024cites this paper
• Orexin neurons mediate temptation-resistant voluntary exercise

  2024cites this paper
• Hypocretin in the nucleus accumbens shell modulates social approach in female but not male California mice

  2024cites this paper
• Single-Cell Profiling Uncovers Evolutionary Divergence of Hypocretin/Orexin Neuronal Subpopulations

  2024cites this paper
• Applications of Lightsheet Fluorescence Microscopy by High Numerical Aperture Detection Lens

  2024cites this paper
• Contribution of hypothalamic orexin (hypocretin) circuits to pathologies of motivation

  2024cites this paper
• Lights, fiber, action! A primer on in vivo fiber photometry

  2023cites this paper
• A multicolor suite for deciphering population coding in calcium and cAMP in vivo

  2023cites this paper
• Targeting Orexin Receptors for the Treatment of Insomnia: From Physiological Mechanisms to Current Clinical Evidence and Recommendations

  2023cites this paper
• Fluorescent proteins and genetically encoded biosensors.

  2023cites this paper
• Understanding Neuropeptide Transmission in the Brain by Optical Uncaging and Release.

  2023cites this paper
• Dopamine neurons in the ventral tegmental area modulate REM sleep.

  2023cites this paper
• Optical tools for visualizing and controlling human GLP-1 receptor activation with high spatiotemporal resolution

  2023cites this paper
• Within-Mice Comparison of Microdialysis and Fiber Photometry-Recorded Dopamine Biosensor during Amphetamine Response

  2023cites this paper
• Illuminating the brain-genetically encoded single wavelength fluorescent biosensors to unravel neurotransmitter dynamics

  2023cites this paper
• Development of a genetically encoded sensor for probing endogenous nociceptin opioid peptide release

  2023cites this paper
• Past, present, and future of tools for dopamine detection.

  2023cites this paper
• Orexin recruits non‐selective cationic conductance and endocannabinoid to dynamically modulate firing of caudal pontine reticular nuclear neurones

  2023cites this paper
• Sensitive multicolor indicators for monitoring norepinephrine in vivo

  2023cites this paper
• Current and emerging methods for probing neuropeptide transmission.

  2023cites this paper
• A genetically encoded sensor for visualizing leukotriene B4 gradients in vivo

  2023cites this paper
• Unilateral Optogenetic Stimulation of Lhx6 Neurons in the Zona Incerta Increases REM sleep.

  2023cites this paper
• System-wide mapping of peptide-GPCR interactions in C. elegans

  2023cites this paper
• Deficiency of orexin signaling during sleep is involved in abnormal REM sleep architecture in narcolepsy

  2023cites this paper
• Spatial memory requires hypocretins to elevate medial entorhinal gamma oscillations.

  2023cites this paper
• Current Status and Future Strategies for Advancing Functional Circuit Mapping In Vivo

  2023cites this paper
• Optogenetic Microwell Array Screening System: A High-Throughput Engineering Platform for Genetically Encoded Fluorescent Indicators

  2023cites this paper
• A tool kit of highly selective and sensitive genetically encoded neuropeptide sensors

  2023cites this paper
• Fiber photometry-based investigation of brain function and dysfunction

  2023cites this paper
• Control and coding of pupil size by hypothalamic orexin neurons

  2022cites this paper
• Activity of GABA neurons in the zona incerta and ventral lateral periaqueductal grey is biased towards sleep.

  2022cites this paper
• A photocaged orexin-B for spatiotemporally precise control of orexin signaling

  2022cites this paper
• Orexin cells efficiently decode blood glucose dynamics to drive adaptive behavior

  2022cites this paper
• System-wide mapping of neuropeptide-GPCR interactions in C. elegans

  2022cites this paper
• Automating the High-Throughput Screening of Protein-Based Optical Indicators and Actuators.

  2022cites this paper
• Within-animal comparison of microdialysis and fiber photometry in amphetamine-exposed mice

  2022cites this paper
• Emerging approaches for decoding neuropeptide transmission.

  2022cites this paper
• GESIAP: A Versatile Genetically Encoded Sensor-based Image Analysis Program

  2022cites this paper
• Recognition and Evaporation Monitoring of Micro-Droplet VOCs Based on CVD-Grown Monolayer MoS2 through a Dielectric Screening Effect

  2022cites this paper
• New Technologies to Investigate Neuropeptides at Scale.

  2022cites this paper
• Role of the locus coeruleus and basal forebrain in arousal and attention.

  2022cites this paper
• Peptidomic Approaches and Observations in Neurodegenerative Diseases

  2022cites this paper
• Coding of reward uncertainty and probability by orexin neurons

  2022cites this paper
• Regulation of Neural Circuitry under General Anesthesia: New Methods and Findings

  2022cites this paper
• Neuropeptide System Regulation of Prefrontal Cortex Circuitry: Implications for Neuropsychiatric Disorders

  2022cites this paper
• Opto-MASS: a high-throughput engineering platform for genetically encoded fluorescent sensors enabling all-optical in vivo detection of monoamines and opioids

  2022cites this paper
• Advancements in the Quest to Map, Monitor, and Manipulate Neural Circuitry

  2022cites this paper
• When the Locus Coeruleus Speaks Up in Sleep: Recent Insights, Emerging Perspectives

  2022cites this paper
• A Molecular Landscape of Mouse Hippocampal Neuromodulation

  2022cites this paper
• Research progress and prospects of genetically encoded neuromodulator sensors

  year unknowncites this paper