Joseph Henry Laboratories of Physics, Princeton University, Princeton, NJ 08544, USA.
Phys Biol. 2012 Oct;9(5):055004. doi: 10.1088/1478-3975/9/5/055004. Epub 2012 Sep 25.
Precise formation of morphogen gradients is essential to the establishment of reproducible pattern in development. Mechanisms proposed for obtaining the requisite precision range from simple models with few parameters to more complex models involving many regulated quantities. The synthesis-diffusion-degradation (SDD) model is a relatively simple model explaining the formation of the Bicoid gradient in Drosophila melanogaster, in which the steady-state characteristic length of the gradient is determined solely by the rates of diffusion and degradation of the morphogen. In this work, we test the SDD model in unfertilized D. melanogaster eggs, which contain a single female pronucleus and lack the nuclear division cycles and other zygotic regulatory processes seen in fertilized eggs. Using two-photon live imaging as well as a novel method for quantitative imaging based on decorrelation of photoswitching waveforms, we find that the Bicoid gradient is longer and shallower in unfertilized eggs as compared to the gradient at the same time points in fertilized eggs. Using a means of measuring the Bicoid lifetime by conjugation to a photoconvertible fluorophore, we find that the lifetime is correspondingly longer in unfertilized eggs, providing qualitative and quantitative agreement with the predictions of the SDD model.
精确形成形态发生梯度对于在发育中建立可重复的模式至关重要。所提出的获得所需精度的机制范围从具有少数参数的简单模型到涉及许多调节数量的更复杂模型。合成-扩散-降解(SDD)模型是一个相对简单的模型,用于解释果蝇中 Bicoid 梯度的形成,其中梯度的稳态特征长度仅由形态发生物质的扩散和降解速率决定。在这项工作中,我们在未受精的果蝇卵中测试了 SDD 模型,这些卵仅含有一个雌性原核,并且缺乏受精卵中所见的核分裂周期和其他合子调节过程。使用双光子活成像以及一种基于光开关波形去相关的定量成像新方法,我们发现与受精卵中相同时间点的梯度相比,未受精卵中的 Bicoid 梯度更长且更浅。通过与光可转化荧光团缀合来测量 Bicoid 寿命的方法,我们发现未受精卵中的寿命相应地更长,与 SDD 模型的预测具有定性和定量的一致性。