Two-Photon Photoactivation of Rubi-Gaba for Studying the Role of the Antisecretory Factor in the Modulation of the GABAA Receptor in Rat Cerebellar Granule Cells In Vitro

Caged compounds, widely diffused in neurophysiology, are molecules biologically or functionally inert until a short pulse of light enables them. Illumination releases the caged effector allowing the activation, manipulation, and control of selective biological functions and variations. in particular, 3D two-photon excitation microscopy, coupled with the electrophysiological technique of the patch-clamp in whole-cell configuration, provides a useful tool for studying several neurobiological processes such as the localized, accurate, and specific receptor response of a well-defined neuronal area. Here, we report this method to figure out the role of the protein Antisecretory Factor (AF) in the modulation of the GABA A receptor by using the caged neurotransmitter RuBi-GABA, rapidly photoreleased, in femtoliter volumes, in a precise and confined region of a neuron at a defined concentration. AF acts in vivo by inhibiting …