Effective sensory communications require massive interconnected interhemispheric cortico-cortical projections. However, little is known at present regarding the exact interactions that occur bilaterally between sensory cortices across the brain hemispheres and how they influence cortical processing. Here, we employ optogenetic stimulation of the whisker-related thalamic excitatory neurons in combination with the somatosensory forepaw stimulation. We demonstrate a novel platform to investigate the interhemispheric interactions underlying cortical sensory processing.
Animal preparation: 3μl AAV5-CaMKIIα::ChR2(H134R)-mCherry was injected to the right VPM of adult rats (200-250g, SD strain, male, n = 6). Four weeks after injection, an opaque optical fiber cannula (d=450μm) was implanted at the injection site as a means to deliver optical stimulation.
Optogenetic and Forepaw Stimulation: Blue light (473nm) was presented to animals expressing ChR2 with a 4-pulse paradigm (10ms pulse width, interstimulus interval, ISI=125ms or 50ms, 40mW/mm2). To map out the responses evoked by VPM optogenetic stimulation, blue light was presented once every 30s, and repeated 20 times. Two needle electrodes were subcutaneously inserted into the right forepaw: one between the first and second digits and the other between the third and fourth digits. The electrodes were fixed using surgical tape and the stimulus effectiveness was first qualitatively confirmed by digit twitching. Electrical stimulation was given via a constant voltage stimulator at 8V with a 4Hz square wave and 3ms pulse duration in a block design paradigm (5s ON, 25s OFF) to evoke robust BOLD responses in primary somatosensory cortex (S1). To investigate the effects of optogenetic stimulation of VPM on the forepaw somatosensory processing, the optogenetic and forepaw stimulation were paired with one another.
fMRI Acquisition and Analysis: fMRI data was acquired at 7T using GE-EPI (FOV=32×32mm2, matrix=64×64, α=56°, TE/TR=20/1000ms, and 10 contiguous slices with 1mm thickness). Data were preprocessed before applying GLM and coherence analysis to identify significant BOLD responses (P<0.001).
BOLD activations evoked by either optogenetic stimulation or forepaw electrical stimulation: 4-pulse optogenetic stimulation at the VPM thalamocortical excitatory neurons activated the ipsilateral primary somatosensory cortex (S1). Optogenetic stimulation with the ISI of 125ms evoked a stronger BOLD response (2.5% vs 1%) with a prolonged BOLD profile when compared to the 4-pulse OG stimulation with an ISI of 50ms. Moreover, BOLD activation in contralateral S1 barrel field (S1BF) was observed when the ISI was 125ms, but not 50ms. As expected, electrical stimulation at forepaw evoked robust positive BOLD response in the contralateral forelimb region of S1 (S1FL).
Optogenetically-evoked contralateral S1 activation suppress the forepaw evoked response: When optogenetic stimulation was presented together with the forepaw electrical stimulation, we observed that the responses in S1FL was significantly reduced only when the ISI of optogenetic stimulation was 125ms, and not 50ms. This suggests the optogenetic stimulation with an ISI of 125ms is capable of evoking BOLD response at the contralateral S1BF, which could likely influence the somatosensory activation in S1FL evoked by forepaw stimulation.
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