ZFIN Image: Xu et al., 2015, Fig. 3

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Fig. 3

Embryonic/Larval and Adult Microglia in Zebrafish CNS Arise from Distinct Origins

(A) A timeline shows the experimental setup. The RBI-, VDA-, and PBI-region-labeled fish were raised and collected at 4 dpf, 15 dpf, 1.5 months, 3 months, and 1 year for analysis.

(B) Immunohistochemistry showed that the RBI- but not VDA- and PBI-region-derived cells (coro1a-GFP⁺ cells) constitute the major population of microglia (Lcp1⁺ cells) in the brains of 4-dpf zebrafish (dorsal view, rostral to the left, a part of the optic tectum).

(C) Immunohistochemistry showed that the VDA- but not RBI- and PBI-derived cells (coro1a-GFP⁺ cells) constitute the major population of microglia (Lcp1⁺ cells) in the brains of 3-month-old adult zebrafish (sectioned brain).

(D) Quantification of the contribution of the RBI-, VDA-, and PBI-region -derived microglia in the developing and adult zebrafish brain. For the RBI-labeled 4-dpf, 15-dpf, 1.5-month, 3-month, and 1-year adult fish, n = 8, 8, 5, 8, and 10, respectively; for the VDA-region-labeled 4-dpf, 15-dpf, 1.5-month, 3-month, and 1-year adult fish, n = 5, 6, 5, 8, and 9, respectively; for the PBI-labeled 4-dpf, 15-dpf, 1.5-month, and 3-month adult fish, n = 7, 6, 7, and 8 respectively. Error bars represent mean ± SEM. p < 0.05; p < 0.01; p < 0.001.

(E) Immunohistochemistry showed the contribution of the RBI-, VDA-, and PBI-region-derived cells (coro1a-GFP⁺ cells) to microglia (Lcp1⁺ cells) in the sectioned 1.5-month zebrafish spinal cord (indicated by the dashed lines). White arrows indicate the coro1a-GFP⁺ microglia in the spinal cord.

(F) Immunohistochemistry showed the contribution of the RBI-, VDA-, and PBI-region-derived cells (coro1a-GFP⁺ cells) to microglia (Lcp1⁺ cells) in the whole-mount 3-month adult zebrafish spinal cord. White arrows indicate the coro1a-GFP⁺ microglia in the spinal cord.

(G) Quantification of the contribution of RBI-, VDA-, and PBI-region-derived microglia in 1.5-month, 3-month, and 1-year adult zebrafish spinal cord. For the RBI-labeled 1.5-month, 3-month, and 1-year adult fish, n = 6, 12, and 8, respectively; for the VDA-region-labeled 1.5-month, 3-month, and 1-year adult fish, n = 4, 5, and 9, respectively; for the PBI-labeled 1.5-month and 3-month adult fish, n = 5 and 6, respectively. Error bars represent mean ± SEM. p < 0.05; p < 0.01; p < 0.001.