Department of Molecular, Cellular and Integrative Physiology, University of California Los Angeles, Los Angeles, CA 90095, USA.
Rep Prog Phys. 2013 Jul;76(7):072601. doi: 10.1088/0034-4885/76/7/072601. Epub 2013 Jun 14.
Living organisms are made of cells that are capable of responding to external signals by modifying their internal state and subsequently their external environment. Revealing and understanding the spatio-temporal dynamics of these complex interaction networks is the subject of a field known as systems biology. To investigate these interactions (a necessary step before understanding or modelling them) one needs to develop means to control or interfere spatially and temporally with these processes and to monitor their response on a fast timescale (< minute) and with single-cell resolution. In 2012, an EMBO workshop on 'single-cell physiology' (organized by some of us) was held in Paris to discuss those issues in the light of recent developments that allow for precise spatio-temporal perturbations and observations. This review will be largely based on the investigations reported there. We will first present a non-exhaustive list of examples of cellular interactions and developmental pathways that could benefit from these new approaches. We will review some of the novel tools that have been developed for the observation of cellular activity and then discuss the recent breakthroughs in optical super-resolution microscopy that allow for optical observations beyond the diffraction limit. We will review the various means to photo-control the activity of biomolecules, which allow for local perturbations of physiological processes. We will end up this review with a report on the current status of optogenetics: the use of photo-sensitive DNA-encoded proteins as sensitive reporters and efficient actuators to perturb and monitor physiological processes.
生物体由能够通过改变内部状态进而改变外部环境来对外界信号做出响应的细胞组成。揭示和理解这些复杂相互作用网络的时空动态是系统生物学领域的主题。为了研究这些相互作用(理解或建模之前的必要步骤),人们需要开发手段来在时空上控制或干扰这些过程,并在快速时间尺度(<分钟)和单细胞分辨率下监测它们的响应。2012 年,在巴黎举行了一次关于“单细胞生理学”的 EMBO 研讨会(由我们中的一些人组织),以根据最近的发展讨论这些问题,这些发展允许进行精确的时空扰动和观察。这篇综述将主要基于那里报告的研究。我们将首先提出一个非详尽的细胞相互作用和发育途径的例子列表,这些例子可能受益于这些新方法。我们将回顾一些用于观察细胞活性的新工具,然后讨论在光学超分辨率显微镜方面的最新突破,该突破允许超越衍射极限进行光学观察。我们将回顾用于光控制生物分子活性的各种方法,这些方法允许对生理过程进行局部扰动。最后,我们将报告光遗传学的现状:使用光敏感 DNA 编码蛋白作为敏感报告器和有效的致动器来干扰和监测生理过程。