ZFIN Image: Xu et al., 2016, Fig. 4

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Figure Caption

Fig. 4

LPC Promotes the Entry of Microglial Precursors into the Brain

(A) A schematic diagram of the dorsal view of zebrafish head. The red square indicates the region where LPC or ATPγS is injected. Normally the upper half-brain and lower half-brain are injected with LPC or ATPγS and control buffer, respectively.

(B) Dorsal view of the ATPγS-injected brain of Tg(Xla.Tubb:bcl-2;mpeg1:loxP-DsRedx-loxP-GFP) embryos at 4-5 hpi. The upper half-brain is injected with ATPγS and the lower half-brain is injected with control buffer. DsRedx⁺ cells represent microglia. The midbrain is labeled by dashed lines.

(C) Quantification of microglia in ATPγS-injected brain of Tg(Xla.Tubb:bcl-2;mpeg1:loxP-DsRedx-loxP-GFP) embryos at 4-5 hpi. n = 8 for control and ATPγS injection. DsRedx⁺ cells represent microglia. Error bars represent the mean ± SEM.

(D) Dorsal view of the LPC-injected brain of Tg(Xla.Tubb:bcl-2;mpeg1:loxP-DsRedx-loxP-GFP) embryos at 4-5 hpi. The upper half-brain is injected with LPC and the lower half-brain is injected with control buffer. DsRedx⁺ cells represent microglia. White arrows indicate LPC-induced microglia in the brain. The midbrain is labeled by dashed lines.

(E) Quantification of microglia in the LPC-injected brain of Tg(Xla.Tubb:bcl-2;mpeg1:loxP-DsRedx-loxP-GFP) embryos at 4-5 hpi. n = 17 for control and LPC injection. Error bars represent the mean ± SEM. ***p < 0.001.

(F) Quantification shows that the number of microglia in the LPC-injected brain of Tg(Xla.Tubb:bcl-2;mpeg1:loxP-DsRedx-loxP-GFP) embryos is drastically decreased by 21 hpi. DsRedx⁺ cells represent microglia. n = 5. Error bars represent the mean ± SEM. **p < 0.01.

(G) Time-lapse imaging pictures show that the microglia in the LPC-injected brain of Tg(mpeg1:loxP-DsRedx-loxP-eGFP) embryos gradually migrate out of the brain. DsRedx⁺ cells represent microglia. White arrowheads indicate one microglia migrating out of the brain. Dashed lines indicate the midbrain.

(H) Dorsal view images of the midbrain of Tg(mpeg1:loxP-DsRedx-loxP-eGFP) control and gpr132b morphants (MO). DsRedx⁺ cells represent microglia. Dashed lines indicate the midbrain.

(I) Quantification of the number of microglia number in the midbrain of Tg(mpeg1:loxP-DsRedx-loxP-eGFP) control and gpr132b morphants (MO). n = 14 for WT control and n = 15 for MO. Error bars represent the mean ± SEM. ***p < 0.001.

See also Figure S4 and Movie S4.

Figure Data

Expression / Labeling details

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Acknowledgments

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Reprinted from Developmental Cell, 38(2), Xu, J., Wang, T., Wu, Y., Jin, W., Wen, Z., Microglia Colonization of Developing Zebrafish Midbrain Is Promoted by Apoptotic Neuron and Lysophosphatidylcholine, 214-22, Copyright (2016) with permission from Elsevier. Full text @ Dev. Cell