Manganese-enhanced MRI of layer-specific activity in the visual cortex from awake and free-moving rats

Cortical responses to visual stimulation have been studied extensively in the rodent, but often require post-stimulation ex vivo examination of the tissue. Here, we test the hypothesis that visual stimulus-dependent cortical activity from awake and free-moving rats can be encoded following systemically administered MnCl2, and activity subsequently readout using manganese-enhanced MRI (MEMRI), a technique that can be performed without sacrificing the animal. Unanesthetized Sprague-Dawley rats, with or without systemic injection of MnCl2, were maintained for 8 h in either a visually stimulating environment or darkness. To identify vision-dependent changes in cortical activity, animals were anesthetized and cortices were examined by 3D RARE MEMRI. Mean signal intensities in sub-cortical regions (e.g., superior colliculus and the lateral geniculate), and cortical regions (primary and accessory visual cortices) were compared. Cortex linearization was performed to aid in layer-specific signal intensity comparisons. Manganese administration alone globally increased signal intensity in the brain (P < 0.0001). In visually stimulated and unstimulated rats, layer-specific analysis revealed that stimulated rats had on average significantly (P < 0.05) higher signal intensities in layers IV and V of the primary visual cortex, as well as in deeper portions of the superficial superior colliculus, relative to dark adapted rats. Such differences went undetected without layer-specific analysis. We demonstrate, for the first time, the feasibility of layer-specific stimulus-dependant non-invasive MEMRI readout after encoding activity in awake and free moving rats. Future MEMRI studies are envisioned that measure the effects on cortical activity of sensory stimulation, as well as normal development, disease, plasticity, and therapy in longitudinal studies.