Figure 3

Time-lapse imaging from different starting stages reveals distinct morphogenetic failure modes leading to embryo death in zfPRL-3 morphants. Embryos injected with 4 ng zfPRL-3 morpholino (MO) were screened at the 1k-cell stage to exclude dead or severely dysmorphic individuals, immobilized in 1% low-melting agarose, and imaged every 1 min at 28 °C using a stereomicroscope. Representative bright-field frames from time-lapse recordings are shown. Time (min) is indicated relative to the first frame of each sequence (0′). (A–E) Embryo imaged from the sphere stage showing prolonged arrest with progressive blastoderm loosening/dispersion and death by 152 min. (F–J) Embryo imaged from the dome stage showing initial epiboly to ~50% epiboly, followed by stalled progression, abnormal inward constriction, blastoderm–yolk separation, and death by 213 min. (K–O) Embryo imaged from ~50% epiboly showing progression toward ~75% epiboly followed by progressive inward constriction with yolk compression, culminating in yolk rupture and death by 243 min. Scale bars = 250 μm.