Extended Data Fig. 8: Glutamate-induced microglia–neuron interactions.

a, Location of microglia process contacts onto jRCaMP1b-expressing neurons in the whisker barrel cortex of Mg^WT mice (images shown in Fig. 2d). Each data point represents the average percentage of all microglia–neuron contacts quantified in an individual mouse. Data are mean ± s.e.m. n = 6 mice. AIS, axonal initial segment. b, Number of microglia in the FOV during in vivo 2P time-lapse imaging of microglial motilities and neuronal activity in awake Mg^WT and Mg^PTX mice. Data are mean ± s.e.m. n = 8 mice per genotype. Not significant, (P = 0.114) by two-tailed unpaired t-test. c, Correlation analysis of intraneuronal Ca²⁺ accumulation and number of microglia in awake Mg^WT and Mg^PTX mice. n = 8 mice per genotype. R² = 0.0059; deviation from zero = not significant (P = 0.856) by linear regression analysis (dotted red line). d, Gene expression of metabotropic (Grm) and ionotropic (Grin) receptors and glutamate transporters (Slc) in Mg^PTX microglia compared to Mg^WT microglia. Data from microglia from n = 3 mice/genotype. Dotted lines indicate 1.5-fold change threshold levels. ^*P = 0.0245; n.s., not significant by unpaired two-tailed t-test. e, In vivo 2P time-lapse imaging of focal (arrows) glutamate uncaging-induced neuronal Ca²⁺ transients (jRCaMP1b, red) and directed microglia motilities (green) in the cortex of awake Mg^WT mice. Scale bar, 25 µm. Representative images are shown for n = 3 mice.

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