Reconstitution of Morphogen Signaling Gradients in Cultured Cells.
Reconstitution of Morphogen Signaling Gradients in Cultured Cells.
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DOI：
10.1007/978-1-0716-1174-6_4
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Li P
中科院分区：
文献类型：
--
作者：
Kim JS;Pineda M;Li P
多细胞生物的发育依赖于信号信息在空间和时间上的正确建立。被称为形态原的分泌分子在空间中形成浓度梯度，并为生物体内的分化细胞提供位置信息。尽管已经确定了形态发生通路的关键分子成分，但在发育中的胚胎中，形态发生通路的结构和关键参数如何控制信号梯度的性质，如它们的大小、速度和对扰动的鲁棒性，仍然具有挑战性。在细胞培养中重建形态梯度提供了解决这个问题的另一种方法。在这里，我们描述了重建小鼠成纤维细胞中Sonic Hedgehog （SHH）信号梯度的方法。该方案包括形态素发送和接收细胞系的设计，径向和线性梯度的设置，定量延时成像和数据分析。类似的方法有可能应用于其他细胞-细胞通讯途径。
Development of multicellular organisms depends on the proper establishment of signaling information in space and time. Secreted molecules called morphogens form concentration gradients in space and provide positional information to differentiating cells within the organism. Although the key molecular components of morphogen pathways have been identified, how the architectures and key parameters of morphogen pathways control the properties of signaling gradients, such as their size, speed and robustness to perturbations, remains challenging to study in developing embryos. Reconstituting morphogen gradients in cell culture provides an alternative approach to address this question. Here we describe the methodology for reconstituting Sonic Hedgehog (SHH) signaling gradients in mouse fibroblast cells. The protocol includes the design of morphogen sending and receiving cell lines, the set-up of radial and linear gradients, the quantitative time-lapse imaging, and the data analysis. Similar approaches could potentially be applied to other cell-cell communication pathways.