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Fig. 3. PLD1 is required for convergent extension movements. (A) PLD1 is involved in convergent extension movements. DMZ explants were dissected at stage 10.5 and cultured until stage 20. PLD1 MO (40 ng) or PLD1 (300 pg) blocked elongation of DMZ explants and co-injection of PLD1 MO with PLD1 (1 ng) rescued its blockade. (B) Quantification of Fig. 3A. The Y-axis indicates the length/width ratio (L/W) of each DMZs. Error bars denote standard deviations. Asterisk indicates p<0.005 when compared to the uninjected control with PLD1-, PLD1+PLDMO and PLD1 MO- injected explants, respectively. (C) In situ hybridization for Gsc and Xnot. Before the onset of CE movements (st.10.25), PLD1 knockdown could not alter expression of Gsc. Converged and extended expression pattern of Xnot was inhibited in PLD1 morphants at mid-gastrula (st.11.5) and early-neurula (st.13). (D) RT-PCR analysis using primers specific for Chordin (Chd), Gsc, Xbra, and Xnr3. Ornithine decarboxylase (ODC) serves as a loading control. (–) RT, a negative control of no reverse transcriptase. DMZ explants from embryos injected with Control MO (40 ng), PLD1 MO (40 ng), or PLD1 RNA (500 pg) were analyzed at stage 10.5. |
