Beck Cade, Singh Tanya, Farooqi Angela, Venkatesh Tadmiri, Vazquez Maribel
Department of Biomedical Engineering, The City College of New York-CUNY, USA.
Department of Biology, The City College of New York-CUNY, USA.
J Neurosci Methods. 2016 Mar 15;262:32-40. doi: 10.1016/j.jneumeth.2015.12.012. Epub 2015 Dec 29.
The developing visual system in Drosophila melanogaster provides an excellent model with which to examine the effects of changing microenvironments on neural cell migration via microfluidics, because the combined experimental system enables direct genetic manipulation, in vivo observation, and in vitro imaging of cells, post-embryo. Exogenous signaling from ligands such as fibroblast growth factor (FGF) is well-known to control glia differentiation, cell migration, and axonal wrapping central to vision.
The current study employs a microfluidic device to examine how controlled concentration gradient fields of FGF are able to regulate the migration of vision-critical glia cells with and without cellular contact with neuronal progenitors.
Our findings quantitatively illustrate a concentration-gradient dependent chemotaxis toward FGF, and further demonstrate that glia require collective and coordinated neuronal locomotion to achieve directionality, sustain motility, and propagate long cell distances in the visual system.
COMPARISON WITH EXISTING METHOD(S): Conventional assays are unable to examine concentration- and gradient-dependent migration. Our data illustrate quantitative correlations between ligand concentration/gradient and glial cell distance traveled, independent or in contact with neurons.
Microfluidic systems in combination with a genetically-amenable experimental system empowers researchers to dissect the signaling pathways that underlie cellular migration during nervous system development. Our findings illustrate the need for coordinated neuron-glia migration in the Drosophila visual system, as only glia within heterogeneous populations exhibited increasing motility along distances that increased with increasing FGF concentration. Such coordinated migration and chemotactic dependence can be manipulated for potential therapeutic avenues for NS repair and/or disease treatment.
黑腹果蝇发育中的视觉系统为通过微流控技术研究微环境变化对神经细胞迁移的影响提供了一个极佳的模型,因为这个综合实验系统能够对胚胎后的细胞进行直接基因操作、体内观察和体外成像。众所周知,来自成纤维细胞生长因子(FGF)等配体的外源性信号控制着对视觉至关重要的神经胶质细胞分化、细胞迁移和轴突包裹。
当前研究采用微流控装置来研究FGF的受控浓度梯度场如何在有或没有与神经祖细胞进行细胞接触的情况下调节对视觉至关重要的神经胶质细胞的迁移。
我们的研究结果定量地说明了对FGF的浓度梯度依赖性趋化作用,并进一步证明神经胶质细胞需要集体且协调的神经元运动,才能在视觉系统中实现方向性、维持运动性并传播较长的细胞距离。
传统检测方法无法检测浓度和梯度依赖性迁移。我们的数据说明了配体浓度/梯度与神经胶质细胞迁移距离之间的定量相关性,无论神经胶质细胞是否与神经元接触。
微流控系统与易于进行基因操作的实验系统相结合,使研究人员能够剖析神经系统发育过程中细胞迁移背后的信号通路。我们的研究结果表明,在果蝇视觉系统中需要协调的神经元-神经胶质细胞迁移,因为只有异质群体中的神经胶质细胞沿着随着FGF浓度增加而增加的距离表现出增强的运动性。这种协调的迁移和趋化依赖性可用于为神经修复和/或疾病治疗探索潜在的治疗途径。