Multicolor Spinal Cord Neural Imaging

Pain is a complex, multidimensional percept that initiates appropriate protective behaviors by integrating sensory information from the spinal cord with ongoing brain states. To drive new mechanistic understanding of spinal cord physiology in awake, behaving animals, we introduced surgical innovations, a redesigned imaging chamber, and improved experimental and computational methods. These modifications enabled spinal cord imaging for months to over a year. We monitored individual axons, identified a somatotopic map, imaged bilateral noxious stimulus-provoked neural dynamics of spinal cord projection neurons in behaving animals, and observed months-long microglial changes after nerve injury. Then, using the Inscopix nVue LScape module, we conducted simultaneous, multicolor monitoring of both sides of the spinal cord in freely moving mice, allowing us to monitor injury-induced alterations in behavior and to correlate these behaviors with neural activity. Most striking are the profound differences between anesthetized and awake imaging, with the latter revealing spontaneous and movement-related activity, including ipsilateral and contralateral responses to peripheral stimuli. Our studies underscore the significance of recording spinal cord activity, long-term, in the behaving animal.

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