Kinesin Light Chain Involved in Kinesin-1 Driven Cytoplasmic Streaming in Drosophila Oocytes
Physical and Biological Sciences
Saxton Lab
Kinesin-1 is a motor protein responsible for the movement of organelles, such as mitochondria and vesicles, toward the plus ends of microtubules. Two known linker proteins are Kinesin Light Chain (Klc), which helps bind Kinesin to many different cargos, and Milton which has been found to bind Kinesin-1 to Mitochondria. We are studying the linkage of Kinesin to its cargoes by observing a Kinesin-dependent progress called cytoplasmic streaming. At a late stage of Drosophila oocyte development, the surrounding nurse cells, which are connected to the oocyte via cytoplasmic bridges, dump their cytoplasm into the oocyte. When this happens, the oocyte mixes the new cytoplasm into the oocyte by streaming to ensure even distribution within the oocyte. The question my thesis is trying to answer is what is the role of Klc and Milton in cytoplasmic streaming. This was done by observing the effects of Klc and Milton inhibition on ooplasmic streaming through use of time lapse confocal microscopy. This was done by quantifying the velocity of fluorescent yolk endosomes during streaming in oocytes in which Klc or Milton or both were knocked down by RNAi. RNAi knockdown of Klc and Milton in strains was verified through qPCR. The results indicate that Klc is important for ooplasmic streaming and that Milton is not.
