Multiple steps in the localization of bicoid RNA to the anterior pole of the Drosophila oocyte

Abstract

The anterior region of the Drosophila embryonic pattern is determined by a gradient of the bicoid (bcd) protein. The correct formation of this gradient requires the localization of bcd RNA to the anterior pole of the egg. Here we use a wholemount in situ technique to examine the process of bcd RNA localization during oogenesis and embryogenesis. While bcd protein becomes distributed in a gradient that extends throughout the anterior two thirds of the early embryo, bcd RNA remains restricted to a much smaller region at the anterior pole. The difference between these distributions indicates that the shape of the protein gradient must depend to some extent on the posterior movement of the protein after it has been synthesized. Four distinct phases of bcd RNA localization can be distinguished during oogenesis. Between stages 6 and 9 of oogenesis, the RNA accumulates in a ring at the anterior end of the oocyte. During the second phase, in stage 9–10a follicles, the RNA also localizes to the apical regions of the nurse cells, demonstrating that the nurse cells possess an intrinsic polarity. As the nurse cells contract during stages 10b–ll, all of the bcd RNA becomes localized to the cortex at the anterior end of the oocyte. During a final phase that must occur between stage 12 of oogenesis and egg deposition, the RNA becomes localized to a spherical region that occupies a slightly dorsal position at the anterior pole. Mutations in the maternal-effect genes, exuperantia (exu) and swallow (sww), lead to an almost uniform distribution of bcd RNA in the early embryo, while staufen (stau) mutations produce a gradient of RNA at the anterior pole, exu mutations disrupt the second stage of bcd RNA localization during oogenesis, sww mutations disrupt the third, and stau mutations affect the fourth phase.