RESUMEN

The ability to directly measure acetylcholine (ACh) release is an essential step toward understanding its physiological function. Here we optimized the GRABACh (GPCR-activation-based ACh) sensor to achieve substantially improved sensitivity in ACh detection, as well as reduced downstream coupling to intracellular pathways. The improved version of the ACh sensor retains the subsecond response kinetics, physiologically relevant affinity and precise molecular specificity for ACh of its predecessor. Using this sensor, we revealed compartmental ACh signals in the olfactory center of transgenic flies in response to external stimuli including odor and body shock. Using fiber photometry recording and two-photon imaging, our ACh sensor also enabled sensitive detection of single-trial ACh dynamics in multiple brain regions in mice performing a variety of behaviors.

Asunto(s)

Acetilcolina/metabolismo , Técnicas Biosensibles/métodos , Encéfalo/metabolismo , Animales , Animales Modificados Genéticamente , Conducta Animal/fisiología , Colinérgicos/farmacología , Drosophila/genética , Drosophila/metabolismo , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Células HEK293 , Humanos , Ratones , Cuerpos Pedunculados/metabolismo , Neuronas/metabolismo , Corteza Olfatoria/metabolismo , Receptor Muscarínico M3/genética , Receptor Muscarínico M3/metabolismo , Corteza Somatosensorial/metabolismo