In vivo analyses of cytoplasmic transport and cytoskeletal organization during Drosophila oogenesis: characterization of a multi-step anterior localization pathway

Abstract

Anterior patterning of the Drosophila embryo depends on localization of bicoid (bcd) mRNA to the anterior pole of the developing oocyte, and bcd mRNA localization requires both the exuperantia (exu) gene and an intact microtubule cytoskeleton. To gain insight into the mechanism of anterior patterning, we have used time lapse laser scanning confocal microscopy to analyze transport of particles containing a Green Fluorescent Protein-Exu fusion (GFP-Exu), and to directly image microtubule organization in vivo. Our observations indicate that microtubules are required for three forms of particle movement within the nurse cells, while transport through the ring canals linking the nurse cells and oocyte appears to be independent of both microtubules and actin filaments. As particles enter the oocyte, a final microtubule-dependent step directs movement to the oocyte cortex. However, our observations and previous studies suggest that the polarity of the oocyte microtubule network is not in itself sufficient to generate anterior asymmetry, and that additional factors are required to restrict morphogens to the anterior pole. Based on these observations, we propose a multi-step anterior localization pathway.