Does Notch Delta Interact At New Adherens Junction Interface Formed Between The Two Sisters Cells
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1) Does Notch Delta interact at new adherens junction interface formed between the two sister cells?
There is evidence in vertebrates developing brains that Notch activation occurs at the level of adherens junction and it is restricted to the two sister cells after division. Our preliminary observations that sister cells share a stable new adherens junctions interface suggests that Notch Delta interactions may occur at this place. To test whether Notch Delta interact at new adherens junction formed between the two sister cells we will first assess whether Notch receptor is activated in this region. For that we will express Notch receptor 1a and 1b tagged with photoconvertible fluorescent protein Dendra as previously described in drosophila: and mouse: . We will photoconvert Notch Dendra at new adherens junction interface and trace the nuclear translocation of Notch intracellular domain. Accordingly to our initial hypothesis Notch1-Dendra photoconverted at the new adherens junction interface but not elsewhere will translocate to the nucleus. Adherens junction will be labeled by Par3-GFP or –CFP and new adherens junction interface will be monitored by live-imaging as shown in fig..
To complement these experiments we will determine whether Delta Notch interact at new adherens junctions interface by tagging Notch ligands (zDelta 1, 2 and 3) and receptors with non-fluorescent split-GFP fragments (GFP exons- 1-10 and GFP exon 11) (ref). We predict that once Delta and Notch