Pharmaco-fUS: Quantification of pharmacologically-induced dynamic changes in brain perfusion and connectivity by functional ultrasound imaging in awake mice

•fUS imaging estimates the functional connectivity brain networks at very high spatial resolution (100 × 100 × 400 µm) in awake and behaving mice.•Pharmaco-fUS can detect the acute modulatory effects of a drug on functional brain connectivity in awake and behaving mice, through the intact skull.•It enables the calculation of a robust ad-hoc pharmacological score of scopolamine-induced changes in connectivity patterns with a limited number of animals (five mice).•We demonstrate dose-dependent specific effects of central cholinergic modulation of functional connectivity independent from global brain perfusion changes.•This study paves the way to high-throughput and large-scale definition of quantitative biomarkers of brain connectomics alterations.