The dynamics of fluorescently labeled endogenous gurken mRNA in Drosophila.
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 sqd(1) 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.
Jaramillo
Angela M
AM
Howard Hughes Medical Institute, Princeton University, NJ 08544, USA.
Weil
Timothy T
TT
Goodhouse
Joseph
J
Gavis
Elizabeth R
ER
Schupbach
Trudi
T
D008841:Actin Cytoskeleton / Q000187:drug effects; D000818:Animals; D004330:Drosophila / Q000187:drug effects / Q000235:genetics*; D029721:Drosophila Proteins / Q000235:genetics*; D004398:Dyneins / Q000378:metabolism; D036681:Fluorescence Recovery After Photobleaching; D009154:Mutation; D009865:Oocytes / Q000737:chemistry; D009866:Oogenesis / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis*; D016211:Transforming Growth Factor alpha / Q000235:genetics*; D050257:Tubulin Modulators / Q000494:pharmacology
D029721:Drosophila Proteins; D012333:RNA, Messenger; D016211:Transforming Growth Factor alpha; D050257:Tubulin Modulators; C083730:grk protein, Drosophila; D004398:Dyneins