Left-right patterning phenotypes in sea mutants and morphants
| . | . | . | Visceral organ asymmetry (%) . | . | . | . | |||
|---|---|---|---|---|---|---|---|---|---|
| Genotype . | Stage . | n . | SS . | SI . | Heterotaxic . | Abnormal . | |||
| seatg238a siblings (+/+, +/–) | 48 hpf | 426 | 98 | 1 | 1 | – | |||
| seatg238a –/– | 48 hpf | 429 | 89 | 4 | 7 | – | |||
| seafa20r siblings (+/+, +/–) | 48 hpf | 400 | 99 | 1 | – | – | |||
| seafa20r –/– | 48 hpf | 249 | 91 | 6 | 3 | – | |||
| Wild-type uninjected | 48 hpf | 52 | 96 | 4 | – | – | |||
| Wild type + e5i5 sea MO | 48 hpf | 148 | 56 | 22 | 14 | 8 | |||
| Asymmetric gene expression (%) | |||||||||
| L | R | B | None | ||||||
| seatg238a clutch | 20 s | 248 | 93 | 1 | 6 | – | |||
| seatg238a –/– | 20 s | 62 | 82 | 3 | 15 | – | |||
| seafa20r clutch | 20 s | 219 | 93 | 1 | 6 | – | |||
| seafa20r –/– | 20 s | 55 | 87 | 2 | 11 | – | |||
| Wild-type uninjected | 20 s | 74 | 97 | 3 | – | – | |||
| Wild type + e5i5 sea MO | 20 s | 66 | 50 | 33 | 17 | – | |||
| . | . | . | Visceral organ asymmetry (%) . | . | . | . | |||
|---|---|---|---|---|---|---|---|---|---|
| Genotype . | Stage . | n . | SS . | SI . | Heterotaxic . | Abnormal . | |||
| seatg238a siblings (+/+, +/–) | 48 hpf | 426 | 98 | 1 | 1 | – | |||
| seatg238a –/– | 48 hpf | 429 | 89 | 4 | 7 | – | |||
| seafa20r siblings (+/+, +/–) | 48 hpf | 400 | 99 | 1 | – | – | |||
| seafa20r –/– | 48 hpf | 249 | 91 | 6 | 3 | – | |||
| Wild-type uninjected | 48 hpf | 52 | 96 | 4 | – | – | |||
| Wild type + e5i5 sea MO | 48 hpf | 148 | 56 | 22 | 14 | 8 | |||
| Asymmetric gene expression (%) | |||||||||
| L | R | B | None | ||||||
| seatg238a clutch | 20 s | 248 | 93 | 1 | 6 | – | |||
| seatg238a –/– | 20 s | 62 | 82 | 3 | 15 | – | |||
| seafa20r clutch | 20 s | 219 | 93 | 1 | 6 | – | |||
| seafa20r –/– | 20 s | 55 | 87 | 2 | 11 | – | |||
| Wild-type uninjected | 20 s | 74 | 97 | 3 | – | – | |||
| Wild type + e5i5 sea MO | 20 s | 66 | 50 | 33 | 17 | – | |||
hpf, hours post fertilization; SS, situs solitus (wild type), SI situs inversus (reversed); s, somites; L, left; R, right; B, bilateral
We determined visceral organ asymmetry based on the position of the heart,liver and pancreas as described previously(Schottenfeld et al., 2007)(see Fig. S1 in the supplementary material). Heterotaxic refers to a random arrangement of organs; thus, not SS or SI. Abnormal refers to malformation of organs that was observed occasionally in MO injected embryos. Asymmetric gene expression refers to combined numbers for southpaw, lefty1 and lefty2 expression
At 48 hpf, –/– embryos were selected on the basis of the curly tail down phenotype. At 20 s, it is not possible to distinguish mutants from siblings morphologically. All embryos with abnormal asymmetric gene expression at 20 s were genotyped. The number of –/– embryos with left expression was extrapolated from the total value on the basis that mutants make up ∼25% of the clutch