Figure 2
Figure 2. Phenotype of rps19-deficient embryos. (A,B) Forebrain () and eye field (▶) are reduced in morphants. In situ hybridization, pax2a probe, 16 hpf. In situ hybridization was performed on 30 morphant or wild-type embryos. (C,D) At 24 hpf, morphants have small head and eyes, pinched midbrain-hindbrain boundary, and necrotic regions in brain (). A typical morphant embryo is shown. (E) In some morphants, forebrain and eyes are absent. The frequency of this phenotype was 0% to 5%. (F) rps19-deficient embryos are smaller than wild-type. (G) Phenotypic manifestations of rps19 deficiency vary between the progeny of different pairs. Red represents normal looking; yellow, mild malformations; blue, strong malformations; purple, forebrain and eyes are absent; violet, dead. Twenty-five healthy looking morphants from each pair were selected at 6 hpf and compared at 20 hpf. Progeny of 4 pairs are shown as an example. (H,I) The level of apoptosis is higher in morphants. Acridine orange staining, 1 μg/mL, 30 minutes, 20 hpf. A representative embryo from at least 30 is shown. (J-L) Cartilage of the head is malformed in morphants. Alcian blue, 120 hpf. (M-O) The morphant fish shown in panel N has edema (); eyes are present but abnormal. The fish shown in panel O has no eyes and forebrain. Note an abnormally large ventral fin in this fish, 120 hpf. (P,Q) Heart field's position and orientation are reversed in some morphants. In situ hybridization, lft2 probe, 19 hpf. An embryo representative of the group of 30 embryos used for in situ hybridization is shown. (R-T) Position of visceral organs is also randomized in morphants. Pancreas is located on the right side in wild-type fish (R), whereas in some morphants it is on the left (S) or bilateral. (T) 120 hpf, insulin probe. (U) rps19-deficient embryos are rescued by injection of 200 pg of rps19 mRNA. The proportion of normal-looking embryos among RPS19-deficient morphants is higher in the group injected with the RNA; the proportion of embryos with severe defects is reduced as is the death rate. No effect was observed after injection of GFP mRNA: 48 hpf, 3.4 ng MOs/embryo. Red represents morphologically normal embryos; yellow, mild defects; blue, severe defects; black, death. Eighty embryos were used per group. Shown are the results representative of 3 independent experiments.

Phenotype of rps19-deficient embryos. (A,B) Forebrain () and eye field (▶) are reduced in morphants. In situ hybridization, pax2a probe, 16 hpf. In situ hybridization was performed on 30 morphant or wild-type embryos. (C,D) At 24 hpf, morphants have small head and eyes, pinched midbrain-hindbrain boundary, and necrotic regions in brain (). A typical morphant embryo is shown. (E) In some morphants, forebrain and eyes are absent. The frequency of this phenotype was 0% to 5%. (F) rps19-deficient embryos are smaller than wild-type. (G) Phenotypic manifestations of rps19 deficiency vary between the progeny of different pairs. Red represents normal looking; yellow, mild malformations; blue, strong malformations; purple, forebrain and eyes are absent; violet, dead. Twenty-five healthy looking morphants from each pair were selected at 6 hpf and compared at 20 hpf. Progeny of 4 pairs are shown as an example. (H,I) The level of apoptosis is higher in morphants. Acridine orange staining, 1 μg/mL, 30 minutes, 20 hpf. A representative embryo from at least 30 is shown. (J-L) Cartilage of the head is malformed in morphants. Alcian blue, 120 hpf. (M-O) The morphant fish shown in panel N has edema (); eyes are present but abnormal. The fish shown in panel O has no eyes and forebrain. Note an abnormally large ventral fin in this fish, 120 hpf. (P,Q) Heart field's position and orientation are reversed in some morphants. In situ hybridization, lft2 probe, 19 hpf. An embryo representative of the group of 30 embryos used for in situ hybridization is shown. (R-T) Position of visceral organs is also randomized in morphants. Pancreas is located on the right side in wild-type fish (R), whereas in some morphants it is on the left (S) or bilateral. (T) 120 hpf, insulin probe. (U) rps19-deficient embryos are rescued by injection of 200 pg of rps19 mRNA. The proportion of normal-looking embryos among RPS19-deficient morphants is higher in the group injected with the RNA; the proportion of embryos with severe defects is reduced as is the death rate. No effect was observed after injection of GFP mRNA: 48 hpf, 3.4 ng MOs/embryo. Red represents morphologically normal embryos; yellow, mild defects; blue, severe defects; black, death. Eighty embryos were used per group. Shown are the results representative of 3 independent experiments.

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