Fig. 4: Spatial resolution of in vivo sonogenetic therapy in V1 cortical neurons. | Nature Nanotechnology

Fig. 4: Spatial resolution of in vivo sonogenetic therapy in V1 cortical neurons.

From: Ectopic expression of a mechanosensitive channel confers spatiotemporal resolution to ultrasound stimulations of neurons for visual restoration

Fig. 4

a, Image of a rat brain expressing G22S MscL–tdTomato (red) in V1. b, Confocal stack projection of a sagittal brain slice expressing G22S MscL–tdTomato (red) and labelled with anti-NeuN antibody (green) and DAPI (blue). The layers of V1 are delineated by the dashed white lines. Magnification of layer 4 of V1 (lower right). c, Densities of NeuN-positive, MscL-positive and double-labelled cells for three brain slices. d, Schematic of the setup used for in vivo electrophysiological recordings and US stimulation. µEcoG electrode array placed on V1 of an MscL-transfected rat (top right). e, Visual-evoked cortical potentials in response to a 100 ms flash (left). Sonogenetic-evoked potentials for 15 MHz US stimuli of various durations (middle). Absence of US responses in an NT rat to a 15 MHz stimulus (right). The black traces represent the mean evoked potentials over 100 trials, individually illustrated by the grey traces. The black arrows indicate the stimulus onsets. f, Duration of sonogenetic µEcog responses for the stimuli of different durations (10 ms, n = 58 trials; 20 ms, n = 32 trials; 50 ms, n = 56 trials on 6 animals). g–i, N1 peak amplitudes for increasing US pressure (g), increasing duration (h) and increasing frequency (i) (n = 6 rats). j,k, Pseudocolour activation maps for the stimuli of increasing US pressure (j) and for a horizontal displacement of the US transducer by 0.8 mm (k) (the arrow indicates the direction of displacement). Each black dot represents an electrode of the array. The colour bar represents the N1 peak amplitude in microvolts. l, Mean activated areas for various US pressure values (n = 6 animals). m, Relative displacement of the activation centre to the previous position following movement of the US transducer by 0.4 mm. Here p = 1 × 10–12, one-sample two-tailed t-test, n = 37 positions on 6 animals (mean, 0.29 ± 0.16 mm (s.d.)). Data are presented as mean values ± s.e.m. Scale bars, 200 and 50 μm (b); 300 μm (j and k).

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