Analysis of pMad and Medea Expression in BMP Pathway in Drosophila with Multiple Fluorescent Proteins
Authors: Hung-Yuan (Zeke) Chen; Gregory Reeves
Affiliation: Texas A&M University
Keywords: m. morphogens; y. live imaging
Morphogen gradients are important in early Drosophila embryo development. One such gradient, the BMP/Dpp gradient, patterns the dorsal region of the embryo, which induces a series of protein interactions. After BMP-like morphogen Dpp transports to the dorsal region and binds to type I receptors Thickveins (Tkv), Tkv is activated by type II receptor Punt and then phosphorylates Mothers against Dpp (Mad). The phosphorylated Mad (pMad) and Smad4 homolog Medea translocate together to the nucleus to regulate the target gene. Compared to the Dpp gradient, pMad and Medea are procedurally closer to gene regulation. However, the transport and expression of pMad and Medea are rarely discussed. Their expression depends on the morphogen concentration which is determined by the cell location. In this work, we tag eGFP and mScarlet I to Mad and Medea and mtagBFP to Histone 2A for locating the nuclei using Crispr-Cas9 techniques and analyze the embryo images after injection into flies and crossing. Finally, we then do image analysis of 14-cycle embryos to see the expression of these proteins.