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Fig. S1
Neuronal injuries and the formation of Ca2+ waves, related to Figure 3
(A1 and A2) Dorsal views of a 3 dpf larval brain, showing neurons (NBT::DsRed) before (A1) and after (A2) injury. (B and C) Histological sections of an un-injured (B) and injured brain (C). The lesion is marked with a white arrowhead. (D) Dorsal view of a 3 dpf larval brain showing microglia in green (pU1::Gal4-UAS-TagRFP) and the vasculature in blue (Fli::eGFP). (E1 and E2) Time lapse of Ca2+ signaling (beta-actin::GCaMP3.1) in a Thapsigargin injected brain before (E1) and after (E2) laser ablation (white arrowhead). (F) Microglia (pU1::Gal4-UAS-GFP) engulf neurons (NBT::DsRed) as shown by the red neuronal material present within the cell. (G) P2Y12 morphant microglia engulf red neuronal material as control cells. (H1 and H2) Time lapse of Ca2+ signaling (beta-actin::GCaMP3.1) in a P2Y12-Mo injected embryo before (H1) and after (H2) laser ablation (white arrowhead). (I1-I3) Time lapse of the microglial response (pU1::Gal4-UAS-GFP) towards bacteria (E. coli) in a P2Y12-Mo injected embryo (Mov.S5). (J) Percentage of microglia in a 100 μm wide circle around the side of injection having engulfed bacteria in control and P2Y12 injected embryos 1 min and 90 min post infection. Error bars represent standard deviations. (K) RT-PCR showing the splice effect of the Duox-Mo. (L1 and L2) Time lapse of Ca2+ signaling (beta-actin::GCaMP3.1) in a CBX injected brain before (L1) and after (L2) laser ablation (white arrowhead). (M1 and M2) microglia (pU1::Gal4-UAS-GFP) before (M1) and 120 min after (M2) central brain injury (white arrowhead) in CBX injected brains. Scale bars always: 20 μm. Images (A, D-H, L, M) were done using an Olympus FV 1000 with a 40X/NA1.15 objective. Images (I1-I3) were done using an Andor Spinning Disk Confocal with a 20x/NA0.7 objective. Images (B, C) were done using a Zeiss Axiophot with a 20X/NA0.5 objective.
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Reprinted from Developmental Cell, 22(6), Sieger, D., Moritz, C., Ziegenhals, T., Prykhozhij, S., and Peri, F., Long-Range Ca(2+) Waves Transmit Brain-Damage Signals to Microglia, 1138-1148, Copyright (2012) with permission from Elsevier. Full text @ Dev. Cell