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Fig. 4 The puf-A gene plays a role in the PGC development in zebrafish embryonic development.
(A) The vasa antisense riboprobe was used in whole-mount in situ hybridization as a marker to monitor primordial germ-cell (PGC) development in zebrafish. The vasa expression in wild-type (WT) and morphants (MO1 at 5 ng/embryo) at 16 and 20 hpf embryos in dorsal view. Anterior is to the left for 16 hpf and left bottom for 20 hpf. The morphants exhibited prominent abnormalities with either a reduction in PGC numbers (50.4%, n = 125 embryos) or abnormal patterns of migration (34.4%, n = 125) indicating the failure of PGC navigation towards their destined sites. (B) Upper: the construction of puf-A siRNA with nanos 3′ UTR, vasa expression and normal eye size in embryos 2 dpf after injection. The embryos displayed a marked reduction in PGC numbers (80.9%, n = 115 embryos) and abnormal patterns of migration (11.3%, n = 115 embryos). Middle: construction of puf-A siRNA without nanos 3′ UTR, vasa expression and small eye size in 2dpf embryos. Bottom: construction of control siRNA with nanos 3′ UTR, vasa expression and normal eye size in embryos after injection.