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Fig. 5. SRp38 inhibits neurogenesis and induces Delta/Id3. (A) In situ hybridization for non-specific ß-tubulin stains the ciliated epidermis and neurons. Left, control embryo. Middle, injection of 500 pg SRp38 targeted to the neural plate results in a loss of neuronal ß-tubulin. Right, overexpression of 500 pg SRp38 in the epidermis leads to a decrease in ciliated epidermal cells. Both of these phenotypes are symptomatic of Notch activation. Injection sites marked by red arrowhead. (B) Schematic model of SRp38 inhibition of neurogenesis. SRp38 inhibition of neurogenin activity may act via Delta and Id3. (C) Lateral views of stage 18 embryos stained for Delta. Left: control embryo. Right: injection of 500 pg of SRp38 induces robust expression of Delta (red arrowhead). (D) RT-PCR analysis of animal caps expressing 500 pg SRp38 (lane 4). (Uninjected control: C, lane 3.) SRp38 induces ectopic expression of Delta, lane 4. EF1 is a loading control and muscle actin (MA) controls for mesodermal contamination. (E) Lateral views of stage 18 embryos stained for Id3. Left: control embryo. Right: injection of 500 pg of SRp38 induces expression of Id3 (red arrowhead). (F) RT-PCR analysis of Id3 splicing. Primers were designed to span exons 1-2 or exons 2-3. Embryos treated with decreasing doses of SRp38 (500 pg to 100 pg) were analyzed for changes in the amount of spliced products. No discernible changes were found.

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