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              <p>The P-glycoprotein (P-gp/ABCB1) is a major efflux transporter which impedes the brain delivery of many drugs across the blood-brain barrier (BBB). Focused ultrasound with microbubbles (FUS) enables BBB disruption, which immediate and delayed impact on P-gp function remains unclear. Positron emission tomography (PET) imaging using the radiolabeled substrate [$^{11}$C]metoclopramide provides a sensitive and translational method to study P-gp function at the living BBB.A FUS protocol was devised in rats to induce a substantial and targeted disruption of the BBB in the left hemisphere. BBB disruption was confirmed by the Evan's Blue extravasation test or the minimally-invasive contrast-enhanced MRI. The expression of P-gp was measured 24 h or 48 h after FUS using immunostaining and fluorescence microscopy. The brain kinetics of [$^{11}$C]metoclopramide was studied by PET at baseline, and both immediately or 24 h after FUS, with or without half-maximum P-gp inhibition (tariquidar 1 mg/kg). In each condition (n = 4–5 rats per group), brain exposure of [11C]metoclopramide was estimated as the area-under-the-curve (AUC) in regions corresponding to the sonicated volume in the left hemisphere, and the contralateral volume. Kinetic modeling was performed to estimate the uptake clearance ratio (R1) of [$^{11}$C] metoclopramide in the sonicated volume relative to the contralateral volume.In the absence of FUS, half-maximum P-gp inhibition increased brain exposure (+135.0 ± 12.9%, p &lt; 0.05) but did not impact R$_1$ (p &gt; 0.05). Immediately after FUS, BBB integrity was selectively disrupted in the left hemisphere without any detectable impact on the brain kinetics of [$^{11}$C]metoclopramide compared with the baseline group (p &gt; 0.05) or the contralateral volume (p &gt; 0.05). 24 h after FUS, BBB integrity was fully restored while P-gp expression was maximally down-regulated (−45.0 ± 4.5%, p &lt; 0.001) in the sonicated volume. This neither impacted AUC nor R1 in the FUS + 24 h group (p &gt; 0.05). Only when P-gp was inhibited with tariquidar were the brain exposure (+130 ± 70%) and R$_1$ (+29.1 ± 15.4%) significantly increased in the FUS + 24 h/tariquidar group, relative to the baseline group (p &lt; 0.001).We conclude that the brain kinetics of [$^{11}$C]metoclopramide specifically depends on P-gp function rather than BBB integrity. Delayed FUS-induced down-regulation of P-gp function can be detected. Our results suggest that almost complete down-regulation is required to substantially enhance the brain delivery of P-gp substrates.</p>
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