The dynamics of fluorescently labeled endogenous gurken mRNA in Drosophila
Author:
Jaramillo Angela M.12, Weil Timothy T.2, Goodhouse Joseph2, Gavis Elizabeth R.2, Schupbach Trudi12
Affiliation:
1. Howard Hughes Medical Institute, Princeton University, NJ 08544, USA 2. Department of Molecular Biology, Princeton University, NJ 08544, USA
Abstract
During Drosophila oogenesis, the targeted localization of gurken (grk) mRNA leads to the establishment of the axis polarity of the egg. In early stages of oogenesis, grk mRNA is found at the posterior of the oocyte, whereas in the later stages grk mRNA is positioned at the dorsal anterior corner of the oocyte. In order to visualize the real-time localization and anchorage of endogenous grk mRNA in living oocytes, we have utilized the MS2-MCP system. We show that MCP-GFP-tagged endogenous grk mRNA localizes properly within wild-type oocytes and behaves aberrantly in mutant backgrounds. Fluorescence recovery after photobleaching (FRAP) experiments of localized grk mRNA in egg chambers reveal a difference in the dynamics of grk mRNA between young and older egg chambers. grk mRNA particles, as a population, are highly dynamic molecules that steadily lose their dynamic nature as oogenesis progresses. This difference in dynamics is attenuated in K10 and sqd1 mutants such that mislocalized grk mRNA in older stages is much more dynamic compared with that in wild-type controls. By contrast, in flies with compromised dynein activity, properly localized grk mRNA is much more static. Taken together, we have observed the nature of localized grk mRNA in live oocytes and propose that its maintenance changes from a dynamic to a static process as oogenesis progresses.
Publisher
The Company of Biologists
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