Development of a genetically encoded sensor for probing endogenous nociceptin opioid peptide release

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: June 25, 2024

Abstract Nociceptin/orphanin-FQ (N/OFQ) is a recently appreciated critical opioid peptide with key regulatory functions in several central behavioral processes including motivation, stress, feeding, and sleep. The functional relevance of N/OFQ action the mammalian brain remains unclear due to lack high-resolution approaches detect this neuropeptide appropriate spatial temporal resolution. Here we develop characterize NOPLight, genetically encoded sensor that sensitively reports changes endogenous release. We characterized affinity, pharmacological profile, spectral properties, kinetics, ligand selectivity, potential interaction intracellular signal transducers NOPLight vitro. Its functionality was established acute slices by exogeneous application chemogenetic induction release from PNOC neurons. In vivo studies fibre photometry enabled direct recording binding exogenous receptor ligands, as well detection within paranigral ventral tegmental area (pnVTA) during natural behaviors activation summary, show here can be used dynamics tissue freely behaving animals.

Language: Английский

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Multiplexed Monitoring of Neurochemicals via Electrografting-Enabled Site-Selective Functionalization of Aptamers on Field-Effect Transistors

Analytical Chemistry, Journal Year: 2022, Volume and Issue: 94(24), P. 8605 - 8617

Published: June 9, 2022

Neurochemical corelease has received much attention in understanding brain activity and cognition. Despite many attempts, the multiplexed monitoring of coreleased neurochemicals with spatiotemporal precision minimal crosstalk using existing methods remains challenging. Here, we report a soft neural probe for neurochemical via electrografting-assisted site-selective functionalization aptamers on graphene field-effect transistors (G-FETs). The probes possess excellent flexibility, ultralight mass (28 mg), nearly cellular-scale dimension 50 μm × each G-FET. As demonstration, show that G-FETs electrochemically grafted molecular linkers (-COOH or -NH2) specific can be used to monitor serotonin dopamine high sensitivity (limit detection: 10 pM) selectivity (dopamine sensor >22-fold over norepinephrine; >17-fold dopamine). In addition, demonstrate feasibility simultaneous single interferences phosphate-buffered saline, artificial cerebrospinal fluid, harvested mouse tissues. stability studies maintain capability simultaneously after incubating rat fluid 96 h, although reduced response at concentrations is observed. Ex vivo mice brains suggest potential applications evoked release serotonin. developed detection methodology also adapted other neurochemicals, such as metabolites neuropeptides, by simply replacing functionalized G-FETs.

Language: Английский

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Sub-100 ms Level Ultrafast Detection and Near-Infrared Ratiometric Fluorescence Imaging of Norepinephrine in Live Neurons and Brains

Journal of the American Chemical Society, Journal Year: 2023, Volume and Issue: 145(43), P. 23832 - 23841

Published: Oct. 18, 2023

Norepinephrine (NE) is a key neurotransmitter in the central and sympathetic nervous systems, whose content fluctuates dynamically rapidly various brain regions during different physiological pathophysiological processes. However, it remains great challenge to directly visualize precisely quantify transient NE dynamics living systems with high accuracy, specificity, sensitivity, and, particular, temporal resolution. Herein, we developed series of small-molecular probes that can specifically detect through sequential nucleophilic substitution–cyclization reaction, accompanied by ratiometric near-infrared fluorescence response, within an impressively short time down 60 ms, which 3 orders magnitude faster than present probes. A unique water-promoted intermolecular proton transfer mechanism disclosed, dramatically boosted recognition kinetics ∼680 times. Benefiting from these excellent features, quantitatively imaged endogenous under external stimuli at single neuron level further revealed close correlations between fluctuations Parkinson's disease pathology acute slices live mouse brains vivo.

Language: Английский

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Advancing social behavioral neuroscience by integrating ethology and comparative psychology methods through machine learning

Neuroscience & Biobehavioral Reviews, Journal Year: 2023, Volume and Issue: 151, P. 105243 - 105243

Published: May 22, 2023

Social behavior is naturally occurring in vertebrate species, which holds a strong evolutionary component and crucial for the normal development survival of individuals throughout life. Behavioral neuroscience has seen different influential methods social behavioral phenotyping. The ethological research approach extensively investigated natural habitats, while comparative psychology was developed utilizing standardized univariate tests. advanced precise tracking tools, together with post-tracking analysis packages, recently enabled novel phenotyping method, that includes strengths both approaches. implementation such will be beneficial fundamental but also enable an increased understanding influences many factors can influence behavior, as stress exposure. Furthermore, future increase number data modalities, sensory, physiological, neuronal activity data, thereby significantly enhance our biological basis guide intervention strategies abnormalities psychiatric disorders.

Language: Английский

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Enzymatic Galvanic Redox Potentiometry for In Vivo Biosensing

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(8), P. 3672 - 3678

Published: Feb. 15, 2024

Redox potentiometry has emerged as a new platform for in vivo sensing, with improved neuronal compatibility and strong tolerance against sensitivity variation caused by protein fouling. Although enzymes show great possibilities the fabrication of selective redox potentiometry, an enzyme electrode to output open-circuit voltage (EOC) fast response remains challenging. Herein, we report concept novel enzymatic galvanic (GRP) time coupling merits high selectivity electrodes excellent biocompatibility reliability GRP sensors. With glucose biosensor illustration, use flavin adenine dinucleotide-dependent dehydrogenase recognition element carbon black potential relay station improve time. We find that rapidly responds good linear relationship between EOC logarithm concentration within range from 100 μM 2.65 mM. The shows over O2 coexisting neurochemicals, reversibility, can monitor dynamics rat brain. believe this study will pave potentiometric biosensing chemical events reliability.

Language: Английский

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