Scanning small spots of laser light allows mapping of synaptic circuits in brain slices from transgenic mice expressing channelrhodopsin-2 (ChR2). The laser spots photostimulate presynaptic neurons expressing ChR2, while postsynaptic responses can be monitored in neurons that do not express ChR2. Correlating the location of the light spot with the amplitude of the postsynaptic response elicited at that location yields maps of the spatial organization of the synaptic circuits. This approach yields maps within minutes, which is several orders of magnitude faster than can be achieved with conventional paired electrophysiological methods. We have applied this high-speed technique to map local circuits in many brain regions. In cerebral cortex, we observed that maps of excitatory inputs to pyramidal cells were qualitatively different from those measured for interneurons within the same layers of the cortex. In cerebellum, we have used this approach to quantify the convergence of molecular layer interneurons on to Purkinje cells. The number of converging interneurons is reduced by treatment with gap junction blockers, indicating that electrical synapses between interneurons contribute substantially to the spatial convergence. Remarkably, gap junction blockers affect convergence in sagittal cerebellar slices but not in coronal slices, indicating sagittal polarization of electrical coupling between interneurons. By measuring limb movement or other forms of behavioral output, this approach also can be used in vivo to map brain circuits non-invasively. In summary, ChR2-mediated high-speed mapping promises to revolutionize our understanding of brain circuitry.

Here we characterize several new lines of transgenic mice useful for optogenetic analysis of brain circuit function. These mice express optogenetic probes, such as enhanced halorhodopsin or several different versions of channelrhodopsins, behind various neuron-specific promoters. These mice permit photoinhibition or photostimulation both in vitro and in vivo. Our results also reveal the important influence of fluorescent tags on optogenetic probe expression and function in transgenic mice.

Studies of the brain promise to be revolutionized by new experimental strategies that harness the combined power of optical techniques and genetics. We have mapped the circuitry of the mouse brain by using both optogenetic actuators that control neuronal activity and optogenetic sensors that detect neuronal activity. Using the light-activated cation channel, channelrhodopsin-2, to locally photostimulate neurons allows high-speed mapping of local and long-range circuitry. For example, with this approach we have mapped local circuits in the cerebral cortex, cerebellum and many other brain regions. Using the fluorescent sensor for chloride ions, Clomeleon, allows imaging of the spatial and temporal dimensions of inhibitory circuits in the brain. This approach allows imaging of both conventional "phasic" synaptic inhibition as well as unconventional "tonic" inhibition. The combined use of light to both control and monitor neural activity creates unprecedented opportunities to explore brain function, screen pharmaceutical agents, and potentially to use light to ameliorate psychiatric and neurological disorders.