Spatiotemporal quantification of subcellular ATP levels in a single HeLa cell during changes in morphology.

Rika Suzuki,Kotaro Oka,Kohji Hotta

doi:10.1038/srep16874

Abstract

The quantitative relationship between change in cell shape and ATP consumption is an unsolved problem in cell biology. In this study, a simultaneous imaging and image processing analysis allowed us to observe and quantify these relationships under physiological conditions, for the first time. We focused on two marginal regions of cells: the microtubule-rich ‘lamella’ and the actin-rich ‘peripheral structure’. Simultaneous imaging and correlation analysis revealed that microtubule dynamics cause lamellar shape change accompanying an increase in ATP level. Also, image processing and spatiotemporal quantification enabled to visualize a chronological change of the relationships between the protrusion length and ATP levels, and it suggested they are influencing each other. Furthermore, inhibition of microtubule dynamics diminished motility in the peripheral structure and the range of fluctuation of ATP level in the lamella. This work clearly demonstrates that cellular motility and morphology are regulated by ATP-related cooperative function between microtubule and actin dynamics.

Highlights

The quantitative relationship between change in cell shape and Adenosine triphosphate (ATP) consumption is an unsolved problem in cell biology
HeLa cells expressing ATeam were imaged under physiological conditions for 10 min, and cytoskeletal dynamics were modulated by 100 nM Latrunculin A or 200 nM Taxol at 3 min
Actin dynamics are reportedly related to ATP levels[10] and we confirmed this assertion through experiments with Latrunculin A (Fig. 1a–c and Fig. 4b,e,i,j)

Summary

Introduction

The quantitative relationship between change in cell shape and ATP consumption is an unsolved problem in cell biology. Synaptic vesicle recycling presents a large ATP burden, which may be because of dynamin that mediates membrane fission[12] These previous reports indicate that variation in ATP levels is related to cellular morphological changes and cytoskeletal dynamics. To demonstrate the presence of a direct relationship under physiological conditions, precise and simultaneous observation of ATP levels and either cellular morphology or cytoskeletal dynamics is necessary. This has been difficult because conventional ATP quantification methods do not allow for high-resolution observation[14]. This technical challenge, we recently successfully investigated the relationship between the motility of the growth cone and the crosstalk of second messengers through a combination of simultaneous imaging with spatiotemporal image processing analysis[15]

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