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Fig. 6. Spatial expression of the ferritin H subunit mRNA in the middle region of the tail in the wild type (a, b) and the mutant (c, d) at stage 48. Whole mount in situ hybridization (WISH) was performed by using sense (a, c) or antisense (b, d) digoxigenin (DIG)-labeled RNA probes. Tadpoles were bleached to remove melanin before hybridization in this experiment. With a sense probe of the ferritin H subunit mRNA, no staining was observed in the tail of both the wild type (a) and the mutant (c) in the negative control. Use of an antisense probe in WISH detected strong staining in the lateral lines (arrowheads) of both the wild type (b) and the mutant (d). In addition, specific expression of the ferritin H subunit mRNA was detected in white pigment cells (d, large arrows), which were present around the dorsal side of the spinal cord (sc) in the mutant (nc notochord). Although melanophores were present around the dorsal side of the spinal cord in the wild type, no staining was observed in melanophores (b). Note that staining was also detected in some epidermal cells (small arrows) in both the wild type (b) and the mutant (d). Bar 100 μm |
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Fig. 7. Expression of the ferritin H subunit mRNA in white pigment cells but not in melanophores. Photographs of the posterior region of the wild type tail (a-d) and the mutant tail (e-h) at stage 48. WISH was performed with sense (a, b, e, f) or antisense (c, d, g, h) DIG-labeled RNA probes (sc spinal cord). Tadpoles were bleached after BM purple staining. The same fields before (a, c, e, g) and after (b, d, f, h) bleaching are shown. In the negative control, with a sense probe of the ferritin H subunit mRNA, no staining was observed in the tail of the wild type (a, b) or the mutant (e, f). Before bleaching, dendritic black melanophores (a) were distinguished from punctate white pigment cells (e), which appeared brown under transmitted light. Melanin was bleached effectively in both melanophores (b) and white pigment cells (f). Staining with an antisense probe indicated that white pigment cells (g, h, arrows), but not melanophores (c, d), expressed the ferritin H subunit mRNA. Note that staining was also detected in the dorsal longitudinal anastomosing vessel (asterisks) in both the wild type (c, d) and the mutant (g, h). Bar 100 μm |
