Fig. 2

Differential vascular expression of CYP genes under basal and BNF-induced conditions.

(A) Stereomicroscope images of whole mount ISH for cyp1a1 and cyp1b1 in 52 hpf WT embryos after 4 hrs exposure to DMSO (A) or 1 uM BNF (B). Strong upregulation of cyp1a1 in skin can be detected. cyp1b1 is also induced in the skin, but to a lesser extent. Numbers indicate embryos with indicated expression pattern/total embryos analyzed. Scale bar is 500 um. (C). Camera lucida drawing of 48 hpf zebrafish embryo from Kimmel, et al 1995 [36]. Dashed lines indicate plane of images to visualize central arteries (CtAs, red) and hindbrain vessels (blue). Confocal image shows dorsal view of hindbrain vasculature of live embryo at 48 hpf (scale bar is 200 um). Cartoon schematic depicts arrangement of hindbrain vessels. (D-I) High magnification images of basal cyp1a1/b1 expression at 52 hpf in the hindbrain (D, G) and CtAs (E, H) of the head vasculature in embryos treated with DMSO. Schematics in F,I summarize hindbrain expression patterns of both genes. (J-O) High magnification images of BNF-induced cyp1a1/b1 expression at 52 hpf in the hindbrain (J, M) and CtAs (K, N) of the head vasculature in embryos treated with 1 uM BNF. Schematics in L,O summarize hindbrain expression patterns of both genes. Analysis of cyp1a1 expression was performed in ahr2 mutant embryos to permit visualization of hindbrain vasculature. Yellow arrowheads mark CtAs. Scale bar is 100 um in all high magnification images. Abbreviations–AHR: aryl hydrocarbon receptor, BA: basilar artery, BCA: basal communicating artery, BNF: beta-naphthoflavone, CtA: central artery, CYP: cytochrome p450, DMSO: dimethylsulfoxide, hpf: hours post fertilization, ISH: in situ hybridization, PCS: posterior communicating segments, PHBC: primordial hindbrain channel, WT: wildtype.