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Fig. 3
Neural crest migration in vivo requires Tks5.
(A–B) Whole mount in situ hybridizations to detect neural crest cells were performed on control (Tks5 MM) and Tks5 morphant (Tks5 MO) embryos at 26 hpf. (A) Neural crest specific riboprobes against foxd3, sox10, and crestin (ctn) were used. Bars indicate anterior-posterior area of migrating cells. (*) indicates an increase in pre-migratory cells compared to controls (B) The number of cells migrating into the trunk region was quantified as described in Materials and Methods. Mean values (n = 18) and SEM were shown in graph. p values obtained from Student′s t-test. ** denotes p<0.01. (C) Control (T5 MM) and Tks5 morphant (T5 MO) Tg(sox10:RFP) embryos (28 hpf) were incubated with acridine orange as a marker for apoptosis and imaged by fluorescence microscopy. NC = neural crest cells, AO = acridine orange, brackets designate similar regions of migrating NC cells, and boxes label similarly positioned individual NC cells in the control and morphant embryos (enlarged in bottom left corner of top panel). (D) Control (T5 MM) and Tks5 morphant (T5 MO) Tg(sox10:RFP) embryos (30 hpf) were analyzed for neural crest migration by confocal time-lapse microscopy for 1.5 hours as described in Materials and Methods. Arrows follow ventral cell migration of an individual cell over the duration. * = protrusions emanating from neural crest cells (D = dorsal, V = ventral, A = anterior, P = posterior). (E) The average velocities of individual neural crest (NC) cells for control (T5 MM)-, Tks5 MO-injected, Tks5myc RNA and Tks5 MO co-injected, or Tks5FFmyc RNA and Tks5 MO co-injected Tg(sox10:RFP) embryos were quantified as detailed in Materials and Methods. Mean values (n = 10) and SEM are shown. p values obtained from Student′s t-test.** denotes p<0.01.