Role of FUS+MB recruited neutrophils in phagocytosis of Aβ

To determine whether neutrophils recruited by FUS+MB BBB treatments assisted in the clearance of Aβ plaques, 7-month-old animals of the TgCRND8 mouse model of AD were administered 3 to 5 biweekly (1 treatment every 2 weeks) MRgFUS+MB BBB treatments, targeted to the bilateral hippocampi 14. MRgFUS+MB BBB treatments were conducted as previously described. The TgCRND8 mouse model is an early onset model of AD that overexpresses a double mutant form of human APP 695 (Swedish (KM670/671NL) + Indiana (V717F) mutations) 33. By 7 months-of-age, TgCRND8 mice exhibit prolific thioflavin S-positive and dense-core plaque deposition, cognitive impairment, activated microglia and astrocytes 34, and synaptic loss 35. Four TgCRND8 animals were given biweekly FUS+MB BBB treatments, and 5 TgCRND8 animals were given sham treatment, consisting of the same preparation and anesthesia exposure but without sonication. Animals were sacrificed on the tenth week, two weeks following the fifth FUS+MB or sham treatment, at 9.5-months-old.

Animals were perfused and brain tissue was harvested as described above. For immunohistochemical analyses, the following primary antibodies were used: anti-mouse Ly6G for neutrophils (127602; BioLegend, San Diego, USA), 6F3D for Aβ plaques (Dako, Glostrup, Denmark), and chicken anti-GFAP for astrocytes (ab4674; Abcam, Cambridge, USA). Five brain sections were analyzed per animal. Stained brain sections were scanned with a confocal microscope and analyzed with FIJI.

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