利用代谢生物传感器观察海胆胚胎发育过程中的代谢调节。

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS
Aidan Furze , Ashley Waldron , Mamiko Yajima
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引用次数: 0

摘要

越来越多的证据表明,代谢调节直接影响细胞的功能和发育,因此可能比以前预期的更加动态。对发育过程中代谢物活动的体内和实时分析对于直接验证这一观点至关重要。在这项研究中,我们采用了两种代谢生物传感器来跟踪海胆早期胚胎发育过程中丙酮酸和氧化磷酸化(Oxphos)的动态变化。丙酮酸传感器 PyronicSF 在有丝分裂装置上显示出信号富集,这与相应的丙酮酸激酶(PKM)的定位模式一致。加入丙酮酸会增加 PyronicSF 信号,而敲除 PKM 会减少其信号,这与细胞中的丙酮酸水平有关。与此类似,读取细胞氧化还原电位的比率计量传感器 Grx-roGFP 也会对 DTT 和 H2O2(已知的 Oxphos 还原剂和诱导剂)做出反应。这些观察结果表明,这些代谢生物传感器忠实地反映了胚胎发生过程中细胞的代谢状态。这些生物传感器的延时成像表明,丙酮酸和 Oxphos 的水平在胚胎发育过程中会发生空间和时间上的变化。在 16 细胞阶段,丙酮酸水平首先在小胚层比其他胚泡增加,并在外胚层保持较高水平,而在胚层内则下降。与此相反,Oxphos 信号在 16 细胞阶段首先在小胚层中降低,而在胃形成过程中则在内胚层中升高,显示出与丙酮酸信号相反的趋势。这些结果表明,在胚胎发生过程中,代谢调控在时间和空间上都是动态的,这些生物传感器是实时监测发育中胚胎代谢活动的宝贵工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Visualizing metabolic regulation using metabolic biosensors during sea urchin embryogenesis

Growing evidence suggests that metabolic regulation directly influences cellular function and development and thus may be more dynamic than previously expected. In vivo and in real-time analysis of metabolite activities during development is crucial to test this idea directly. In this study, we employ two metabolic biosensors to track the dynamics of pyruvate and oxidative phosphorylation (Oxphos) during the early embryogenesis of the sea urchin. A pyruvate sensor, PyronicSF, shows the signal enrichment on the mitotic apparatus, which is consistent with the localization patterns of the corresponding enzyme, pyruvate kinase (PKM). The addition of pyruvate increases the PyronicSF signal, while PKM knockdown decreases its signal, responding to the pyruvate level in the cell. Similarly, a ratio-metric sensor, Grx-roGFP, that reads the redox potential of the cell responds to DTT and H2O2, the known reducer and inducer of Oxphos. These observations suggest that these metabolic biosensors faithfully reflect the metabolic status in the cell during embryogenesis. The time-lapse imaging of these biosensors suggests that pyruvate and Oxphos levels change both spatially and temporarily during embryonic development. Pyruvate level is increased first in micromeres compared to other blastomeres at the 16-cell stage and remains high in ectoderm while decreasing in endomesoderm during gastrulation. In contrast, the Oxphos signal first decreases in micromeres at the 16-cell stage, while it increases in the endomesoderm during gastrulation, showing the opposite trend of the pyruvate signal. These results suggest that metabolic regulation is indeed both temporally and spatially dynamic during embryogenesis, and these biosensors are a valuable tool to monitor metabolic activities in real-time in developing embryos.

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来源期刊
ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
自引率
2.10%
发文量
464
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