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Fig. 1 SI. Design of in vivo and ex vivo experiments of the study of loud music-induced opening of the blood-brain barrier (OBBB): (1) three days before experiments the polyethylene catheter (PE-10 tip, Scientific Commodities Inc., Lake Havasu City, Arizona) was implanted into the femoral vein for injection of tracers (Evans Blue dye, fluorescein isothiocyanate dextran 70 kDa, gadolinium-diethylene-triamine-pentaacetic acid - Gd-DTPA); (2) – the optical window [1] was prepared for in vivo real time fluorescent microscopy of the BBB permeability to the Evans Blue Albumin Complex (EBAC, 68.5 kDa) in awake behavior mice (see session “In vivo real time fluorescent microscopy of extravasation of Evans Blue”); (3) – afterward mice (n=15 in each group) were underwent to the intermittent music (70-90-100 dB, 11-10,000 Hz, Scorpions “Still loving you”) during 2h (60 sec sound and 60 sec – pause) (see session “Experimental design of music effect on the BBB permeability”); (4) – Evans Blue dye was injected via catheter immediately after music-off and then we performed in vivo real time fluorescent microscopy of OBBB for EBAC in awake behavior mice during 5 hrs; (5) The EBAC leakage was detected as bright fluorescence around the cerebral microvessels; (6) after in vivo real time fluorescent microscopy of OBBB for EBAC, all mice were decapitated, their brain removed and analyzed using spectrofluorometric assay of EBAC extravasation (see session “Spectrofluorometric assay of EBAC extravasation”); (7) additionally, magnetic resonance imaging (MRI) was used for the study of the BBB permeability for Gd-DTPA; (8) also ex vivo confocal imaging and in vivo real-time two-photon laser scanning microscopy of FITC-dextran extravasation was performed; (9) the time points for injections of the model compounds in relation to the music intervention and the brain collection.; Fig. 2 SI. The frequency range of music (Scorpions “Still Loving You”): Frequencies in the range of 11-10,000 Hz and maximal intensity around 100 Hz; Fig. 3 SI. The measure of sound level in the cages of animals during listening of loud music. To produce the loud music, we used loudspeaker generated sound 100 dB. The location of cage with mice from loudspeaker was: 1) at 1m - 100 dB; 2) at 2m - 90 dB; 3) at 32m - 70 dB in according with information referred here [https://lgmproducts.com/technical-information/sound-pressure-levels-in-dba]. The sound energy was measured directly in a cage of animals using a sound level meter (Megeon 92130, Russia); Fig. 4 SI. The effect of loud music on the BBB permeability to EBAC (µg/g tissue) in different brain fields: *** - p |
