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Fig. 4. Antisense morpholino inhibition of apelin/APJ signaling in the frog embryo leads to vascular patterning defects. (A) Control experiments were carried out as
previously described (Small et al., 2005) to demonstrate effective inhibition of translation. Briefly, fusion transcripts contained 5′ UTR sequences of the target mRNAs
upstream of the EGFP coding region. Fluorescence was visualized under UVillumination. (B) Embryos were injected with antisense morpholino into one cell of the
two-cell embryo. The uninjected side of the embryo serves as a stage-matched control. In all cases, structure of vascular tissue was assayed by in situ hybridization
using probe for erg transcripts. (B, C) Control and injected side respectively of an embryo treated with apelin antisense morpholino (apelin MO1). Note reduction in
intersegmental vessels on the MO-treated side (dashed arrows), while the majority of vascular structures appear to be unaffected. (D, E) Control and treated side
respectively of an embryo injected with a second apelin antisense MO (apelin MO2), again showing disruption of intersegmental vessels in the injected side. (F, G)
Control and treated side respectively of an embryo injected with APJ antisense MO. Once again, growth of intersegmental vessels is disrupted on the injected side
(dashed arrows). (H) Coinjection of APJ mRNA partially rescues intersomitic defects generated using APJ MO1. Rescue with both 200 and 400 pg of APJ mRNA is
statistically significant using the Chi-squared test. |
