Neuroimaging measurements of functional connectivity are commonly interpreted as an index of reciprocal interareal communication. However, direct testing of this hypothesis has been lacking. Using chemogenetics, electrophysiology and resting-state fMRI in the mouse, we show that acute and chronic silencing of the prefrontal cortex result in paradoxical rsfMRI overconnectivity of the mouse default mode network (DMN) and increased delta activity, an effect relayed to wider cortical territories by polymodal thalamic areas. Our results challenge prevailing interpretations of functional connectivity and implicate a critical contribution of sub-cortical rhythm generators to the establishment of large-scale functional coupling.
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