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Fig. 2. Marked alterations in migratory behaviour are detectable during earliest stages of PGC development. (A) Snapshots from a low magnification time-lapse movie recorded over 4 hours of zebrafish development. The area marked in the top left panel is magnified in the rest of the panels. The germ cells remain in their positions for about 1.5 hours following PGC specification and start to migrate actively around dome stage (4.5 hpf, supplementary material Movie 1, n=4 embryos). An asterisk marks a PGC, which leaves the cell cluster. (B-E) High-magnification snapshots from time-lapse movies of wild-type embryos (see supplementary material Movies 2-5). (B) At 3 hpf, PGCs exhibit simple morphology. (C,D) At 3.7 hpf (C) up to 4.3 hpf (D) the cells develop small, as well as long complex protrusions extended in random directions, yet are not polarized and do not migrate. At these stages, the cells show round morphology during cell divisions (D). (E) Soon after 4.3 hpf, PGCs exhibit polarized elongated cell morphology and extend pseudopodia in the direction of migration. (F) Snapshots from a low magnification 1-hour long time-lapse movie of PGCs from a 12 hours donor embryo in a 3 hpf host embryo. The transplanted cells migrate in the early host (supplementary material Movie 6, three cells in two embryos examined). (G) Transplanted cells similar to those in F show polarized elongated cell morphology and migration behaviour similar to PGCs after 4.3 hpf (supplementary material Movie 7, 40x magnification). The white scale bars represent 10 µm and the white arrowheads indicate the direction of migration.

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