Mapping brain networks in awake mice using combined optical neural control and fMRI. J Neurophysiol 105: 1393-1405, 2011. First published December 15, 2010 doi:10.1152/jn.00828.2010. Behaviors and brain disorders involve neural circuits that are widely distributed in the brain. The ability to map the functional connectivity of distributed circuits, and to assess how this connectivity evolves over time, will be facilitated by methods for characterizing the network impact of activating a specific subcircuit, cell type, or projection pathway. We describe here an approach using high-resolution blood oxygenation level-dependent (BOLD) functional MRI (fMRI) of the awake mouse brain-to measure the distributed BOLD response evoked by optical activation of a local, defined cell class expressing the light-gated ion channel channelrhodopsin-2 (ChR2). The utility of this opto-fMRI approach was explored by identifying known cortical and subcortical targets of pyramidal cells of the primary somatosensory cortex (SI) and by analyzing how the set of regions recruited by optogenetically driven SI activity differs between the awake and anesthetized states. Results showed positive BOLD responses in a distributed network that included secondary somatosensory cortex (SII), primary motor cortex (MI), caudoputamen (CP), and contralateral SI (c-SI). Measures in awake compared with anesthetized mice (0.7% isoflurane) showed significantly increased BOLD response in the local region (SI) and indirectly stimulated regions (SII, MI, CP, and c-SI), as well as increased BOLD signal temporal correlations between pairs of regions. These collective results suggest opto-fMRI can provide a controlled means for characterizing the distributed network downstream of a defined cell class in the awake brain. Opto-fMRI may find use in examining causal links between defined circuit elements in diverse behaviors and pathologies.Functional MRI (fMRI) based on the blood oxygenation leveldependent (BOLD) signal (Kwong et al. 1992;Ogawa et al. 1990Ogawa et al. , 1992) is widely used to indirectly measure neural activity in distributed brain networks in humans and nonhuman primates. A recent study has shown that optogenetic strategies, using activation of channelrhodopsin-2 (ChR2)-expressing neurons (Boyden et al. 2005), can be used with high-field MRI imaging to evoke BOLD signals in the anesthetized rodent (Lee et al. 2010). We present data obtained using optogenetic activation of a specific cell class in the awake animal, mapping and characterizing the distributed network responses that result. These opto-fMRI studies collectively open up a wide array of opportunities for exploring the relation between BOLD responses and neural activity and to use rodent models to assess the impact of causal manipulations on distributed brain networks.The recent paper of Lee et al. (2010) These results raise the possibility that opto-fMRI could serve as a tool to explore properties of distributed brain networksfor example, enabling researchers to focus their electrophysiolo...