Zipper Is Necessary for Branching Morphogenesis of the Terminal Cells in the Drosophila melanogaster’s Tracheal System

SIMPLE SUMMARY: The Drosophila melanogaster, also commonly known as the fruit fly, has a relatively simple structure, allowing scientists to study its anatomy. This research was carried out to investigate how a protein called Zipper may be important for the development of the model organism during the early developmental stages. The study concentrated on the respiratory system, also known as the tracheal system, more specifically the leading cells in the tracheal system also known as terminal cells. Zipper was shown to be in the cytoplasm of terminal cells, indicating that it may function in the D. melanogaster’s tracheal system. Then, comparisons between normal fruit flies and those engineered so that the RNA for zipper does not function were made. Visual and quantitative comparisons demonstrated less branching of the terminal cells for the mutants, while no differences were found for lumenogenesis—tube formation within the branched structures. Therefore, this study demonstrates the role of Zipper in branching of the terminal cells in the D. melanogaster’s tracheal system. This study adds onto the existing scientific literature by demonstrating the role of a specific protein in an important biological process occurring in most living organisms. ABSTRACT: Branching morphogenesis and seamless tube formation in Drosophila melanogaster are essential for the development of vascular and tracheal systems, and instructive in studying complex branched structures such as human organs. Zipper is a myosin II’s actin-binding heavy chain; hence, it is important for contracting actin, cell proliferation, and cell sheet adhesion for branching of the tracheal system in post-larval development of the D. melanogaster. Nevertheless, the specific role of Zipper in the larva is still in question. This paper intended to investigate the specific role of Zipper in branching morphogenesis and lumenogenesis in early developmental stages. It did so by checking the localization of the protein in the cytoplasm of the terminal cells and also by analyzing the morphology of zipper RNAi loss-of-function mutants in regard to branching and lumen formation in the terminal cells. A rescue experiment of RNAi mutants was also performed to check the sufficiency of Zipper in branching morphogenesis. Confocal imaging showed the localization of Zipper in the cytoplasm of the terminal cells, and respective quantitative analyses demonstrated that zipper RNAi terminal cells develop significantly fewer branches. Such a result hinted that Zipper is required for the regulation of branching in the terminal cells of D. melanogaster. Nevertheless, Zipper is not significantly involved in the formation of seamless tubes. One hypothesis is that Zipper’s contractility at the lateral epidermis’ leading edge allows cell sheet movement and respective elongation; as a result of such an elongation, further branching may occur in the elongated region of the cell, hence defining branching morphogenesis in the terminal cells of the tracheal system.