Swelling along brain's axons may be true culprit in Alzheimer's disease yale nature
Eight-month-old 5xFAD mice were anaesthetized with ketamine/xylazine solution and hair was removed from the skull area. Buprenex , dexamethasone and carprofen were given subcutaneously at this point. The mouse was placed on a heating pad during the surgery and anaesthesia was checked periodically. Povidone-iodine solution was applied to the skin and cleaned with ethanol, and eye ointment was applied to the eyes.
. We injected 10 µl of Aβ solution into each mouse and collected the brain the next day. Brain tissue was prepared for immunohistochemistry analysis.Six-to-eight-month-old 5xFAD mice were injected with GCaMP6 virus through the subarachnoid space on one hemisphere to label cortical neurons and measure local axonal conduction properties. For measurements of interhemispheric axonal conduction, GCaMP6f virus was injected stereotaxically with the following coordinates: bregma ).
The ROI was identified under a two-photon microscope. GCaMP6-labelled neurons were imaged through excitation at a wavelength of 920 nm. A limited field of view was used to improve the sampling rate. GCaMP6s was imaged at 2 Hz and GCaMP6f was imaged at 10 to 20 Hz. Only axons that displayed spontaneous calcium transients at least once per minute were used for analysis. For stimulated calcium events, mice were anaesthetized using 0.5% isoflurane.
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