Extended Data Fig. 8: Prolonged, broad two-photon optogenetic activation of glutamatergic neurons dilates penetrating arterioles, depending on aSMC’s NMDA receptors.
(a) Schematic graph of the experimental workflow of broad 2 P optogenetics (b) Two-photon image of a p-arteriole (labeled with Hydrazide Alex 633, white arrows) at a cortical depth of 128 μm and surrounding ChR2-mCherry positive glutamatergic axons (green arrows). (c) Still frame images of p-arterioles before and during photostimulation in the littermate control (Grin1f/f) and aSMC-cKOGrin1 mice (SMACreER:Grin1f/f). The GCamp6s intensity further enhanced in the same axons when illumination power switched from 25 mW (white arrow) to 80 mW laser (green arrow). The white dashed lines outline the basal arteriole calibers. (d) Time course of changes in p-arteriolar diameter in response to 1100-nm laser at 25 mW and the following 960-nm laser with increasing power in c. (e) Maximum dilation amplitudes in c (magenta, n = 8 p-arterioles from 6 mice) or cistern magna injection of D-AP5 (100 μM, green, n = 4 p-arterioles from 3 mice) in control mice or aSMC-cKOGrin1 mice (orange, n = 6 p-arterioles from 5 mice). Data are shown as the mean ± s.e.m.; nested, one-way ANOVA after a post hoc Bonferroni multiple comparison adjustments (e).
