Fig. 2.

Effects of pank2 morpholino during zebrafish development. Standard (ST-MO) and pank2 morpholino (P2-MO) were micro-injected at 1/2-cell stage (1 pmol/embryo) and the phenotype observed at 24 and 48 hpf. At both developmental stages, morphants showed abnormalities in the brain region, presence of hydrocephalus (*) and heart edema (≠) (D), severe perturbation of main vessels (dorsal aorta and caudal vein) and vascular plexus in the tail (arrowhead and magnification B’, D’). E) Semi-quantitative RT-PCR analysis of pank2 transcript levels in non-injected (CNT), ST-MO- and P2-MO-injected embryos at 24 and 48 hpf. F) Densitometric analysis of RT-PCR products resolved by agarose-gel electrophoresis. * = P < 0.001. G) Quantitative analysis of the phenotype observed in P2-MO-injected embryos at 48 hpf. Two different phenotypes could be distinguished: a severe one (12%) with head malformations, presence of hydrocephalus and evident edema in the caudal plexus, severe defects in anterior-posterior axis and delay in development; a milder phenotype (59%) with not well-defined brain areas, less severe hydrocephalus and edema in caudal plexus. The graph shows the analysis performed in six independent experiments on a total of 562 injected embryos. Abbreviations: hv, hindbrain ventricle, h, hindbrain, m, midbrain, mhb, midbrain–hindbrain boundary.