The scatterplot colormap indicates the pixel density. reduction of the phosphorylated tau. Conclusion: Findings offered herein reinforce the premise of ultrasound in reducing tau pathology and thus curbing the progression of Alzheimer’s disease. and are the pixel intensities of the reddish and green channel respectively while and are the mean background values of the reddish and green channel respectively. The equations describing the coefficients were implemented in MATLAB. Following the isolation of the CA1 sector from the surrounding tissue, the dependency coefficients were calculated using the natural intensities as inputs. Pixels outside the region of interest were assigned the arithmetic representation for Not-a-Number (NaN) that does not affect the calculation of the coefficients. Statistical analysis All values are expressed as means standard error of the mean (SEM). Differences between the means of the sham and treated groups as well as the ipsilateral and contralateral to the sonication sides were analyzed using 2-way repeated steps ANOVA. Statistical analysis was performed around the mean value obtained per animal; however, all samples (2-3 slices per mouse brain) are offered in the bar plots included in the study. Longitudinal analysis of the BBB opening volume and differences in the means distributed over the hippocampal subfields were analyzed with multiple Student’s t assessments. The P values in both analyses were Pidotimod adjusted based on the Holm Sidak post hoc correction. All statistical analyses were performed using Prism 8 (Graphpad Software, San Diego, CA, USA) and the null hypothesis was rejected at the 0.05 level. Throughout the manuscript the F value in respect to the associated degrees of freedom is provided with the P value adjusted to the corrected multiple t-tests. The significance levels correspond to: Pidotimod * P<=0.05, ** P<=0.01, *** P<=0.0005, **** P<0.0001. Results FUS-induced BBB opening did not differ in volume between groups and across weeks Confirmation of the BBB Pidotimod opening and volume assessment was performed prior to the immunohistochemical analysis to evaluate potential differences in the opening volume. Representative coronal planes of the BBB opening at Bregma -2.70 0.56 are shown in Physique ?Physique22 for each group per week of sonication. Quantification of the opening volume (transgenic; wild-type, week 1: 54.8 0.9 mm3; 52.83 3.9 mm3, week 2: 52.46 2.9 mm3; 54.98 2.6 mm3, week 3: 54.22 3.3 mm3; 55.48 1.6 mm3, week 4: 52.5 2.5 mm3; 54.49 4.3 mm3) did not reveal any significant differences in the opening size across weeks suggesting that repeated ultrasound application did not compromise the integrity of the barrier at least at this age (3.5-4.5 months old). Moreover, the comparable BBB opening volumes of transgenic to wild-type animals indicated a similar response of the BBB to the ultrasound without evidence of edematous incidences as indicated by the T2-weighted images (Physique S3). Open in a separate window Physique 2 Volumetric analysis of the blood-brain barrier opening from your contrast-enhanced T1-weighted MR coronal images. The volumes of the transgenic animals were at the order of 54.8 2.02 mm3, 52.46 6.59 mm3, 54.22 7.44 mm3 and 52.5 5.64 mm3, while 52,83 8.81 mm3, 54.98 5.97 mm3, 55.48 3.75 mm3and 54.49 9.76 mm3 of the wild-type mice Cd200 for the four consecutive weeks. Longitudinal analysis did not show any significant difference across weeks. Additionally, the opening volumes did not differ between transgenic and wild-type animals within the same week interval. FUS-induced BBB opening does not compromise neuronal integrity FUS applied within the security regime has been proven beneficial 26,35,53 while vascular incidences have been reported in sonication protocols including overlapping regions and repeated applications 43. Despite operating in a safe ultrasound parametric space evaluated in previous studies 33,53, we were interested in determining any potential effects of the technique around the neuronal integrity. Therefore, two slices per transgenic brain were counterstained for neuronal cells with the anti-NeuN (neuron-specific nuclear protein) and anti–tubulin antibodies (neuron-specific microtubule element) along with brain slices from a wild-type.