Abstract

Optogenetic stimulation of the mouse cortex can be used to generate motor maps that are similar to maps derived from electrode-based stimulation. Here we present a refined set of procedures for repeated light-based motor mapping in ChR2-expressing mice implanted with a bilateral thinned-skull chronic window and a chronically implanted electroencephalogram (EEG) electrode. Light stimulation is delivered sequentially to over 400 points across the cortex, and evoked movements are quantified on-line with a three-axis accelerometer attached to each forelimb. Bilateral maps of forelimb movement amplitude and movement direction were generated at weekly intervals after recovery from cranial window implantation. We found that light pulses of ~2 mW produced well-defined maps that were centered approximately 0.7 mm anterior and 1.6 mm lateral from bregma. Map borders were defined by sites where light stimulation evoked EEG deflections, but not movements. Motor maps were similar in size and location between mice, and maps were stable over weeks in terms of the number of responsive sites, and the direction of evoked movements. We suggest that our method may be used to chronically assess evoked motor output in mice, and may be combined with other imaging tools to assess cortical reorganization or sensory-motor integration.

Highlights

  • Optogenetic stimulation is a well-suited method for studying the motor system in rodents

  • Light-induced cortical depolarization can be monitored through the chronic EEG electrode, while evoked forelimb movements are quantified on-line with lightweight accelerometers attached to the forelimbs

  • STABILITY OF CHRONIC WINDOW PREPARATION We found that the mice tolerated the bilateral cranial windows without noticeable discomfort or health complications, similar to the original polished and reinforced thinned-skull” (PoRTS) preparation (Drew et al, 2010)

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Summary

Introduction

Optogenetic stimulation is a well-suited method for studying the motor system in rodents. During light-based motor mapping (LBMM), the cortical region representing forelimb muscles can be identified by sequentially stimulating different cortical points and recording the evoked electromyographic (EMG) activity from the contralateral forelimb (Ayling et al, 2009; Hira et al, 2009) This method complements previously available tools, such as intracortical microstimulation (ICMS) where an electrode is lowered into deep cortical layers and limb or body movements are evoked through current injection (Asanuma and sakata, 1967; Li and Waters, 1991; Kleim et al, 1998; Monfils et al, 2005; Tennant et al, 2011; Young et al, 2011; Barbay et al, 2012). Relatively few studies have performed longitudinal motor cortex stimulation (Harrison et al, 2012), and none with bilateral stimulation in combination with electroencephalogram (EEG) recording

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