Figure 1
Figure 1. Zebrafish MCs structurally and functionally resemble their mammalian counterparts. H&E staining. (A) Intestine. (B) Gill. Periodic acid-Schiff (PAS). (C) Intestine. (D) Gill. Toluidine blue. (E) Intestine. (F) Gill. MCs in each panel indicated by ▶. Transmission electron microscope (Phillips 300; FEI, Hillsboro, OR) images of (G) a zebrafish intestinal MC (20 000× magnification) and (H) a mouse bone marrow–derived MC (26 000× magnification). Zebrafish MCs demonstrate a positive reaction to (I) a polyclonal antibody raised against human CD117 (C-KIT) antigen (Dako Cytomation, Carpinteria, CA) and (J) a monoclonal anti–human MC tryptase antibody (gills; Dako Cytomation). Biotinylated Universal Linker (Dako Cytomation) secondary antibody and 3,3′-diaminobenzidine for chromogenic detection (hematoxylin counterstain; 100× objective). RNA ISH using digoxigenin-labeled RNA antisense probe to zebrafish cpa5 demonstrates positive staining in (K) intestinal MCs and (L) pancreas; MCs indicated by ▶. Adult zebrafish injected intraperitoneally with 10 μg of compound 48/80 demonstrate (N) MC degranulation compared with (M) saline-injected controls. PAS staining, 100× objective; MCs in each panel indicated by ▶. (O) Increased plasma tryptase levels compared with saline-injected control fish. Presented as means plus SEM of 3 experiments with 4 to 6 fish per group; *P < .05 (t test). Cytospin of FACS-sorted, FITC-labeled, Fast Red–stained cpa5+ cells isolated from zebrafish embryos at 7 dpf demonstrate morphology consistent with MCs. (P) Toluidine blue. (Q) Wright-Giemsa. (R) Green channel (FITC). (S) Red channel (Fast Red; also Figure S2). (A-F, I-N) Images obtained at 100×/1.3 NA oil-immersion objective with a Nikon Eclipse E600, Nikon DXM 1200 camera, and ACT-1 software (all Nikon, Tokyo, Japan). Panels assembled using Adobe Photoshop CS3 Extended version 10.0 (Adobe Systems, San Jose, CA). (P-S) Images obtained at 100×/1.4 NA oil immersion objective with a Zeiss Axioplan 2 (Zeiss, Jena, Germany) and Axiocam HRc digital camera, Axiovision 4.6 with multi-channel fluorescence module (Diagnostic Instruments, Sterling Heights, MI).

Zebrafish MCs structurally and functionally resemble their mammalian counterparts. H&E staining. (A) Intestine. (B) Gill. Periodic acid-Schiff (PAS). (C) Intestine. (D) Gill. Toluidine blue. (E) Intestine. (F) Gill. MCs in each panel indicated by ▶. Transmission electron microscope (Phillips 300; FEI, Hillsboro, OR) images of (G) a zebrafish intestinal MC (20 000× magnification) and (H) a mouse bone marrow–derived MC (26 000× magnification). Zebrafish MCs demonstrate a positive reaction to (I) a polyclonal antibody raised against human CD117 (C-KIT) antigen (Dako Cytomation, Carpinteria, CA) and (J) a monoclonal anti–human MC tryptase antibody (gills; Dako Cytomation). Biotinylated Universal Linker (Dako Cytomation) secondary antibody and 3,3′-diaminobenzidine for chromogenic detection (hematoxylin counterstain; 100× objective). RNA ISH using digoxigenin-labeled RNA antisense probe to zebrafish cpa5 demonstrates positive staining in (K) intestinal MCs and (L) pancreas; MCs indicated by ▶. Adult zebrafish injected intraperitoneally with 10 μg of compound 48/80 demonstrate (N) MC degranulation compared with (M) saline-injected controls. PAS staining, 100× objective; MCs in each panel indicated by ▶. (O) Increased plasma tryptase levels compared with saline-injected control fish. Presented as means plus SEM of 3 experiments with 4 to 6 fish per group; *P < .05 (t test). Cytospin of FACS-sorted, FITC-labeled, Fast Red–stained cpa5+ cells isolated from zebrafish embryos at 7 dpf demonstrate morphology consistent with MCs. (P) Toluidine blue. (Q) Wright-Giemsa. (R) Green channel (FITC). (S) Red channel (Fast Red; also Figure S2). (A-F, I-N) Images obtained at 100×/1.3 NA oil-immersion objective with a Nikon Eclipse E600, Nikon DXM 1200 camera, and ACT-1 software (all Nikon, Tokyo, Japan). Panels assembled using Adobe Photoshop CS3 Extended version 10.0 (Adobe Systems, San Jose, CA). (P-S) Images obtained at 100×/1.4 NA oil immersion objective with a Zeiss Axioplan 2 (Zeiss, Jena, Germany) and Axiocam HRc digital camera, Axiovision 4.6 with multi-channel fluorescence module (Diagnostic Instruments, Sterling Heights, MI).

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