Fig. 3: Mitochondrial distribution is stable during early phases of microglial response to focal tissue injury.

A Representative image from multiphoton live imaging of NAc microglia (TdTomato) and their mitochondria (mitoGFP) in an acute brain slice from young adult MG-mitoGFP;Ai14 mouse prior to induction of focal laser injury. B TdTom+ microglia and GFP+ mitochondria from the same field of view as in A immediately following induction of focal laser lesion (T0). Ring 1 encompasses the primary lesion area, with inner and outer rings at 15 μm and 30 μm from ring 1, respectively. C–E TdTom+ microglia and GFP+ mitochondria from the same field of view as in A at 5, 10, and 20 min post laser lesions. F, G Percent change in TdTomato signal (microglial processes) within the outer and inner rings, respectively, relative to T0. One-way ANOVA with repeated measures: Outer ring within subjects comparison F_(3,24) = 1.6, P = 0.215, Outer ring between subjects comparison F_(1,8) = 0.01, P = 0.942, Inner ring within subjects comparison F_(3,24) = 2.38, P = 0.095, Inner ring between subjects comparison F_(1,8) = 4.08, P = 0.078. H, I Percent change in GFP signal (mitochondria) within the outer and inner rings, respectively, relative to T0. One-way ANOVA with repeated measures: Outer ring within subjects comparison F_(3,24) = 1.2, P = 0.318, Outer ring between subjects comparison F_(1,8) = 3.69, P = 0.091, Inner ring within subjects comparison F_(3,24) = 5.4, P = 0.005, Inner ring between subjects comparison F_(1,8) = 14.5, P = 0.005. For data in F–I, N = 3 NAc acute brain sections from 3 mice (9 NAc acute brain sections total) and data was plotted as mean +/- SEM. Mice used for these experiments were not treated with 4-hydroxytamoxifen. Source data used to generate all graphs are provided as a Source Data file.