Cortical movement of Bicoid in early Drosophila embryos is actin- and microtubule-dependent and disagrees with the SDD diffusion model

PLoS One. 2017 Oct 3;12(10):e0185443. doi: 10.1371/journal.pone.0185443. eCollection 2017.

Abstract

The Bicoid (Bcd) protein gradient in Drosophila serves as a paradigm for gradient formation in textbooks. The SDD model (synthesis, diffusion, degradation) was proposed to explain the formation of the gradient. The SDD model states that the bcd mRNA is located at the anterior pole of the embryo at all times and serves a source for translation of the Bicoid protein, coupled with diffusion and uniform degradation throughout the embryo. Recently, the ARTS model (active RNA transport, synthesis) challenged the SDD model. In this model, the mRNA is transported at the cortex along microtubules to form a mRNA gradient which serves as template for the production of Bcd, hence little Bcd movement is involved. To test the validity of the SDD model, we developed a sensitive assay to monitor the movement of Bcd during early nuclear cycles. We observed that Bcd moved along the cortex and not in a broad front towards the posterior as the SDD model would have predicted. We subjected embryos to hypoxia where the mRNA remained strictly located at the tip at all times, while the protein was allowed to move freely, thus conforming to an ideal experimental setup to test the SDD model. Unexpectedly, Bcd still moved along the cortex. Moreover, cortical Bcd movement was sparse, even under longer hypoxic conditions. Hypoxic embryos treated with drugs compromising microtubule and actin function affected Bcd cortical movement and stability. Vinblastine treatment allowed the simulation of an ideal SDD model whereby the protein moved throughout the embryo in a broad front. In unfertilized embryos, the Bcd protein followed the mRNA which itself was transported into the interior of the embryo utilizing a hitherto undiscovered microtubular network. Our data suggest that the Bcd gradient formation is probably more complex than previously anticipated.

MeSH terms

  • Actins / metabolism*
  • Animals
  • Drosophila / embryology*
  • Drosophila Proteins
  • Embryo, Nonmammalian / metabolism
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism*
  • Hypoxia
  • Microtubules / drug effects
  • Microtubules / metabolism*
  • Models, Biological*
  • RNA, Messenger / genetics
  • Trans-Activators / genetics
  • Trans-Activators / metabolism*
  • Vinblastine / pharmacology

Substances

  • Actins
  • Drosophila Proteins
  • Homeodomain Proteins
  • RNA, Messenger
  • Trans-Activators
  • bcd protein, Drosophila
  • Vinblastine

Grants and funding

This work was supported by the Swedish Research Council (https://www.vr.se/) (Stefan Baumgartner: study design, data collection and analysis, decision to publish, or preparation of the manuscript); Eric Philipp Sörensen Stiftelse (Stefan Baumgartner: study design, data collection and analysis, decision to publish, or preparation of the manuscript); Byggmästare Olle Engkvist Stiftelse (Stefan Baumgartner: study design, data collection and analysis, decision to publish, or preparation of the manuscript); Russian Foundation for Basic Research (Alexander Spirov: study design, data collection and analysis). A.V.S. is partially supported by RFBR grants 15-04-07800 and 15-04-06480. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.