Journal Article

A genetically encoded sensor for in vivo imaging of orexin neuropeptides

Dernic, Jan, Adamantidis, Antoine R., Tyagarajan, Shiva K., Figueiredo, Marta, Ravotto, Luca, Tsai, Yuan-Chen, Duffet, Loïc, Panniello, Mariangela, Patriarchi, Tommaso, Zych, Anna D., Stoeber, Miriam, Fellin, Tommaso, Weber, Bruno, Zhou, Xuehan, Schmidt, Markus H., Radoux-Mergault, Arthur, Gogolla, Nadine, Bracey, Edward, Kosar, Seher, Burdakov, Denis, Viberti, Bianca

2022

Overview

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.

• Publisher: Nature Publishing Group US,Nature Publishing Group
• Subject: 631/1647/1888/2249 / 631/1647/245/2225 / 631/378/548 / Anesthesia / Animals / Behavior, Animal / Bioinformatics / Biological Microscopy / Biological Techniques / Biomedical and Life Sciences / Biomedical Engineering/Biotechnology / Brain - metabolism / Central nervous system / Female / Fluorescence / Genetic code / Green fluorescent protein / HEK293 Cells / Humans / Hypothalamus / Imaging / Life Sciences / Male / Mice / Mice, Inbred C57BL / Molecular Imaging - methods / Neuroimaging / Neuropeptides / Orexin Receptors - genetics / Orexin Receptors - metabolism / Orexins / Orexins - analysis / Orexins - genetics / Orexins - pharmacology / Photometry / Photons / Proteomics / Recombinant Proteins - genetics / Recombinant Proteins - metabolism / Reproducibility of Results / Sensors / Sleep - physiology / Sleep and wakefulness / Somatosensory cortex