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. 2020 Feb 6;10:2072. doi: 10.1038/s41598-020-58832-8

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Flagella of Chlamydomonas paralyzed-flagella (pf) mutants beat at high pressure. (a) Schematic illustration of the Chlamydomonas flagellum in cross section. Mutant flagella lacking the central pair (CP) (blue) or radial spokes (RS) (red) are paralyzed under physiological conditions. Bar: 100 nm. (b) Percentage of motile cells under different pressure conditions. Cells of pf14(Red), pf18(blue), and WT (black) were examined. Mean ± SD for more than 20 cells were examined in WT and pf mutants, respectively. Temperature: 25 °C. (c) Typical swimming trajectories of pf cells for three seconds at atmospheric and high pressures. (Upper panel) pf14 lacking radial spokes. (Lower panel) pf18 lacking the central pair. At high pressure, a number of cells randomly swam for short distances, while some cells swam straight (cell a and b). Temperature: 35 °C. (d) (Left) Asymmetric bending pattern in a forward-swimming cell of pf14 (cell a in c). Bending of the two flagella is asynchronous. (Right) Symmetric bending pattern in a backward-swimming cell of pf18 (cell b in c). Bending of the two flagella is synchronous. Cells were observed by high-speed video microscopy. Numbers on the left or right indicate the time after the onset of recording. (e) Time course of the pf18 cell movement after being activated by pressure application. The fraction of moving cells reached a peak within 10 seconds, and then gradually decreased. More than 40 cells were examined for each time point. The time course was similar at 25 and 35 °C. A small percentage of cells continued to swim for more than 3 minutes. (f) Reversibility of the pressure-induced motility in pf14. A high pressure (60 MPa) and the atmospheric pressure (0.1 MPa) were alternately applied each for 30 sec. The number of moving cells 10 sec after the pressure change was shown in the graph. Cells stopped and resumed movements, both within 1 sec after the release from and application of high pressure, respectively. More than 120 cells were examined for each time point. Temperature: 25 °C.