High-throughput three-dimensional chemotactic assays reveal steepness-dependent complexity in neuronal sensation to molecular gradients

Zhen Xu, Peilin Fang, Bingzhe Xu, Yufeng Lu, Jinghui Xiong, Feng Gao, Xin Wang, Jun Fan and Peng Shi ()
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Zhen Xu: City University of Hong Kong
Peilin Fang: City University of Hong Kong
Bingzhe Xu: City University of Hong Kong
Yufeng Lu: City University of Hong Kong
Jinghui Xiong: City University of Hong Kong
Feng Gao: City University of Hong Kong
Xin Wang: City University of Hong Kong
Jun Fan: City University of Hong Kong
Peng Shi: City University of Hong Kong

Nature Communications, 2018, vol. 9, issue 1, 1-11

Abstract: Abstract Many cellular programs of neural development are under combinatorial regulation by different chemoattractive or chemorepulsive factors. Here, we describe a microfluidic platform that utilizes well-controlled three-dimensional (3D) diffusion to generate molecular gradients of varied steepness in a large array of hydrogel cylinders, allowing high-throughput 3D chemotactic assays for mechanistic dissection of steepness-dependent neuronal chemotaxis. Using this platform, we examine neuronal sensitivity to the steepness of gradient composed of netrin-1, nerve growth factor, or semaphorin3A (Sema3A) proteins, and reveal dramatic diversity and complexity in the associated chemotactic regulation of neuronal development. Particularly for Sema3A, we find that serine/threonine kinase-11 and glycogen synthase kinase-3 signaling pathways are differentially involved in steepness-dependent chemotactic regulation of coordinated neurite repellence and neuronal migration. These results provide insights to the critical role of gradient steepness in neuronal chemotaxis, and also prove the technique as an expandable platform for studying other chemoresponsive cellular systems.

Date: 2018
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DOI: 10.1038/s41467-018-07186-x

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