Zebrafish PRL-3 knockdown impairs early embryonic development. (A) Survival of wild-type (WT) embryos (N = 300) and embryos injected with control morpholino (MO) (4 ng, N = 115) or zfPRL-3 MO (2 ng, N = 115; 4 ng, N = 62; 6 ng, N = 100; 8 ng, N = 64) was monitored using a 2.3 nL injection volume. Survival was scored at 4, 6, 8, and 24 h post fertilization (hpf). (B) Distribution of developmental stage categories at 9 hpf from an independent injection experiment. Embryos injected with control MO (N = 69) or zfPRL-3 MO (4 ng, N = 60; 8 ng, N = 83) were scored as sphere, dome to 50% epiboly (the interval from dome stage to 50% epiboly), shield to 75% epiboly (the interval from shield stage to 75% epiboly), 90% epiboly, or dead. Percentages shown on the stacked bars indicate the fraction of embryos in each category within the indicated group. (C–G) Representative bright-field images illustrating morphological categories among embryos that survived to 24 hpf following injection with control MO (4 ng) or zfPRL-3 MO (4 ng). Percentages indicate the proportion of embryos within each indicated group; values are shown as n/N, followed by the percentage (%). (C) Surviving control morphants (N = 76) all reached the prim-5 stage (76/76, 100%). (D–G) In contrast, surviving zfPRL-3 morphants (N = 49) at 24 hpf were classified into distinct morphological categories: (D) embryos with abnormal body-axis morphology (21/49, 42.9%), (E) embryos with apparently normal morphology (13/49, 26.5%), (F) embryos developmentally arrested at or before ~75% epiboly (12/49, 24.5%), and (G) embryos arrested near the completion of epiboly (3/49, 6.1%). Scale bars = 250 μm.
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