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一个盘基网柄菌细胞的亚群表现出对 cAMP 极高的敏感性,以进行定向迁移。

A sub-population of Dictyostelium discoideum cells shows extremely high sensitivity to cAMP for directional migration.

机构信息

Laboratory of Single Molecule Biology, Graduate School of Science and Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka, 565-0871, Japan.

Laboratory for Integrated Biodevice, Center for Biosystems Dynamics Research (BDR), RIKEN, 1-3 Yamadaoka, Suita, Osaka, 565-0871, Japan.

出版信息

Biochem Biophys Res Commun. 2021 May 21;554:131-137. doi: 10.1016/j.bbrc.2021.03.095. Epub 2021 Mar 27.

Abstract

The chemotaxis of Dictysotelium discoideum cells in response to a chemical gradient of cyclic adenosine 3',5'-monophosphate (cAMP) was studied using a newly designed microfluidic device. The device consists of 800 cell-sized channels in parallel, each 4 μm wide, 5 μm high, and 100 μm long, allowing us to prepare the same chemical gradient in all channels and observe the motility of 500-1000 individual cells simultaneously. The percentage of cells that exhibited directed migration was determined for various cAMP concentrations ranging from 0.1 pM to 10 μM. The results show that chemotaxis was highest at 100 nM cAMP, consistent with previous observations. At concentrations as low as 10 pM, about 16% of cells still exhibited chemotaxis, suggesting that the receptor occupancy of only 6 cAMP molecules/cell can induce chemotaxis in very sensitive cells. At 100 pM cAMP, chemotaxis was suppressed due to the self-production and secretion of intracellular cAMP induced by extracellular cAMP. Overall, systematic observations of a large number of individual cells under the same chemical gradients revealed the heterogeneity of chemotaxis responses in a genetically homogeneous cell population, especially the existence of a sub-population with extremely high sensitivity for chemotaxis.

摘要

使用新设计的微流控装置研究了 Dictysotelium discoideum 细胞对环腺苷酸 3',5'-单磷酸(cAMP)化学梯度的趋化性。该装置由 800 个细胞大小的通道组成,每个通道宽 4μm,高 5μm,长 100μm,允许我们在所有通道中制备相同的化学梯度,并同时观察 500-1000 个单个细胞的运动。对于从 0.1 pM 到 10 μM 的各种 cAMP 浓度,确定了表现出定向迁移的细胞的百分比。结果表明,在 100 nM cAMP 时趋化性最高,与以前的观察结果一致。在低至 10 pM 的浓度下,约 16%的细胞仍表现出趋化性,这表明仅 6 个 cAMP 分子/细胞的受体占据就可以诱导非常敏感的细胞发生趋化性。在 100 pM cAMP 时,由于细胞外 cAMP 诱导的细胞内 cAMP 的自我产生和分泌,趋化性受到抑制。总体而言,在相同的化学梯度下对大量单个细胞进行系统观察揭示了遗传同质细胞群体中趋化性反应的异质性,特别是存在对趋化性具有极高敏感性的亚群。

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