Neutrophil number and distribution confirm the presence of inflammation in mecp2-null larvae. (A) Total numbers of Tg(mpx:eGFP)-positive neutrophils were counted in 3, 4 and 5 dpf wild-type and mecp2-null larvae using stereo fluorescent microscopy (n=12 larvae per condition pooled from two individual experiments; larvae were scored for three consecutive days). (B) Representative stereo microscopy images of 4 dpf Tg(mpx:eGFP) wild-type and mecp2-null larvae. (C) Numbers of Tg(mpx:eGFP)-positive neutrophils associated with the GI tract of 2, 3, 4 and 5 dpf wild-type and mecp2-null larvae were counted (n≥12 embryos per condition; data are representative of three individual experiments). (D) Representative confocal micrographs (maximum projection) of the GI tracts of 5 dpf Tg(mpx:eGFP) wild-type and mecp2-null larvae in which the GI tract has been delineated with a white dashed line based on the transmitted light images. (E) Representative confocal micrographs (maximum projection) of the brain region of 3 dpf Tg(mpeg1:eGFP) wild-type and mecp2-null larvae. (F) Brain-associated Tg(mpeg1:eGFP)-positive cells were counted for 3 dpf wild-type, heterozygous and mecp2-null larvae (n=7, n=11, n=6, respectively; one-way ANOVA with Tukey's post hoc test; ns, not significant; data are representative of two individual experiments). (G) Total numbers of Tg(mpeg1:eGFP)-positive cells were counted in 3, 4 and 5 dpf wild-type and mecp2-null larvae using stereo fluorescent microscopy (n=11 and n=12 embryos per condition, respectively; data are representative of two individual experiments). A Student’s t-test was used for all statistical analyses, except for the data analyzed in F, by comparing wild-type and mecp2-null numbers per day (***P<0.001; **P<0.01; *P<0.05; ns, not significant).
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