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To achieve simultaneous measurement of multiple cellular events in molecularly defined groups of neurons in vivo, we designed a spectrometer-based fiber photometry system that allows for spectral unmixing of multiple fluorescence signals recorded from deep brain structures in behaving animals. Using green and red Ca2+ indicators differentially expressed in striatal direct- and indirect-pathway neurons, we were able to simultaneously monitor the neural activity in these two pathways in freely moving animals. We found that the activities were highly synchronized between the direct and indirect pathways within one hemisphere and were desynchronized between the two hemispheres. We further analyzed the relationship between the movement patterns and the magnitude of activation in direct- and indirect-pathway neurons and found that the striatal direct and indirect pathways coordinately control the dynamics and fate of movement.
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•Spectral unmixing of multi-color fluorescence signals in vivo•Versatile, inexpensive, and easy to assemble from off-the-shelf parts•Dual-sensor recordings reveal synchronized activity between dual striatal pathways•Direct and indirect pathways coordinately control the dynamics and fate of movement
Using a new method for simultaneous multi-color fluorescence measurements, Meng et al. show that neural activities are synchronized between two parallel striatal pathways, and the magnitude of activation in these two pathways coordinately determines the dynamics and fate of movement.