Examination of microglia and DC-like cells in myeloid–deficient mutant lines.
A-D. Immunofluorescence on transverse brain sections from Tg(p2ry12:p2ry12-GFP) transgenic adult wild-type (A-D), irf8-/- (E-H), csf1ra-/- (I-L), csf1rb-/- (M-P) and csf1ra-/-; csf1rb-/- (csf1rDM) (Q-T) fish, co-stained with anti-GFP (green) and Lcp1 (magenta) antibodies. A, E, I, M, Q. For each genotype, illustrative case of the merge of the two channels, allowing to discriminate in the parenchyma GFP+; Lcp1+ microglia (white arrowheads) from GFP-; Lcp1+ DC-like cells (yellow arrowheads). Single channels high magnification of the insets (dashed frame) in A (B-D), E (F-H), I (J-L), M (N-P) and Q (R-T). Outline yellow arrowheads indicate the absence of GFP signal in corresponding yellow arrowhead pointed cells. Scale bar in (A), (E), (I), (M) and (Q) represents 100 µm and scale bar in other images 50 µm. U-V. Quantification of the cell density for GFP+ Lcp1+ microglia and GFP-Lcp1+ DC-like cells in the dorsal (U), ventral (V) and whole area (W) of the brain for each genotype (n=3). Data in U-W are mean ± SEM. *P<0.05, **P<0.01 (Klustal-Wallis test with Dunn’s post-hoc).