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. Author manuscript; available in PMC: 2013 Oct 31.
Published in final edited form as: Mol Genet Genomics. 2009 Feb 17;281(5):10.1007/s00438-009-0428-8. doi: 10.1007/s00438-009-0428-8

Figure 4. Overexpression of a guanylate cyclase downstream of a viral insertion results in morphological defects.

Figure 4

A- Gal4-VP16 expression in a gene-trap line where mCherry is under the control of UAS and serves as a marker for expression. Expression is present in all areas of neurogenesis including the retina, brain and spinal cord. B–C Expression of gucy2F at 24 hpf in normal (B) and embryos with the viral insertion upstream of gucy2F and Gal4-VP16 (C). D – Phenotype of mutant larvae at 5 dpf presenting with a shortened trunk, hemorrhaging in the brain and abnormal head morphology particularly in the area of the forebrain. E–F- Coronal sections of a phenotypically normal (E) and abnormal (F) 5 dpf larvae. Many neurons are absent in the forebrain of the abnormal larvae shown as a decrease in mCherry+ neurons. Sections were counterstained in Hoechst 33342 to show nuclei in blue G - Linker-mediated PCR for viral insertion. Lanes 1–4 are abnormal larvae, lanes 5–8 are normal siblings showing one viral insertion appears only in the abnormal larvae. The smaller band in lane 6 is due to incomplete enzymatic digestion and shares no sequence similarity to those bands in lanes 1–4. H - Genomic PCR for the gucy2F viral insertion. Lanes 1–4 are abnormal larvae, lanes 5–7 are normal siblings. The viral insertion upstream of gucy2F is present only in the abnormal larvae. Lower panel is β-actin showing DNA is present in all samples. I – analysis of the overexpressed gucy2F transcript at 24hpf. RNA from mCherry + embryos (lanes 1&2) and mCherry − embryos (lane 3&4) was analyzed. Analysis of the 3′ end reveals a 3 kb band is present only mCherry + embryos. Analysis of the 5′ end indicates two splice variants in mCherry+ embryos that are not present in the mCherry − embryos. Scale Bars are 50 μM.