Flagellum couples cell shape to motility in<i>Trypanosoma brucei</i>

Sun, Stella; Kaelber, Jason T.; Chen, Muyuan; Dong, Xiaoduo; Nematbakhsh, Yasaman; Shi, Jing; Dougherty, Matthew; Lim, Chwee Teck; Schmid, Michael F.; Chiu, Wah; He, Cynthia Y.

doi:10.1073/pnas.1722618115

Cited by 32 publications

(37 citation statements)

References 57 publications

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“…Samples frozen by rapid-freezing or high-pressure-freezing techniques retain their native structure, and 55 tomography allows reconstruction of 3D structures within a slab of tissue. Cryo-ET is particularly useful 56 for examination of native cytoskeletal structure (Jasnin et al, 2013;Turgay et al, 2017;McIntosh et al, 57 2018;Sun et al, 2018). We adapted stereocilia blotting methods (Neugebauer and Thurm, 1984;58 Shepherd et al, 1989;Hasson et al, 1997;Avenarius et al, 2017) to isolate a thin layer of stereocilia on 59 electron microscopy grids.…”

Section: Introduction 28mentioning

confidence: 99%

Electron cryo-tomography of vestibular hair-cell stereocilia

Metlagel

Krey

Song

et al. 2018

Preprint

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18High-resolution imaging of hair-cell stereocilia of the inner ear has contributed substantially to our 19 understanding of auditory and vestibular function. To provide three-dimensional views of the structure of 20 stereocilia cytoskeleton and membranes, we developed a method for rapidly freezing unfixed stereocilia 21 on electron microscopy grids, which allowed subsequent 3D imaging by electron cryo-tomography. 22Structures of stereocilia tips, shafts, and tapers were revealed, demonstrating that the actin paracrystal 23 was not perfectly ordered. This sample-preparation and imaging procedure will allow for examination of 24 structural features of stereocilia in a near-native state. 25 2 Keywords 26 Hair cells; stereocilia; cryo-electron microscopy; actin 27

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Section: Introduction 28mentioning

confidence: 99%

Electron cryo-tomography of vestibular hair-cell stereocilia

Metlagel

Krey

Song

et al. 2018

Preprint

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“…Propensity for waveform initiation at different sites may be related to differences in the molecular composition of outer dynein arms between the proximal and distal axoneme 19 . Alternatively, the higher stiffness of the wider cell body (which is laterally attached to the flagellum towards its base) 29 may simply be too rigid for the reversed beat to bend.…”

Section: Resultsmentioning

confidence: 99%

High-speed multifocal plane fluorescence microscopy for three-dimensional visualisation of beating flagella

Wheeler

2019

Preprint

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Analysis of flagellum beating in three dimensions is important for understanding how cells can undergo complex flagellum-driven motility and the ability to use fluorescence microscopy for such three-dimensional analysis would be extremely powerful. Trypanosoma and Leishmania are unicellular parasites which undergo complex cell movements in three dimensions as they swim and would particularly benefit from such an analysis. Here, high-speed multifocal plane fluorescence microscopy, a technique in which a light path multi-splitter is used to visualise 4 focal planes simultaneously, was used to reconstruct the flagellum beating of Trypanosoma brucei and Leishmania mexicana in three dimensions. It was possible to use either an organic fluorescent stain or a genetically-encoded fluorescence fusion protein to visualise flagellum and cell movement in three dimensions at a 200 Hz frame rate. This high-speed multifocal plane fluorescence microscopy approach was used to address two open questions regarding Trypanosoma and Leishmania swimming: To quantify the planarity of the L. mexicana flagellum beat and analyse the nature of flagellum beating during T. brucei 'tumbling'.

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“…Extensive research has already shown that distinct waveforms and beat frequencies can be produced by the same ciliary structure [16,17] (also Man et al, this Issue), depending on intrinsic and extrinsic forcing [18,19,20]. This ability to modulate dynein motor cooperativity to produce distinct beating modes on a single cilium does not however explain the higher-level coordination over a network of such oscillating structures.…”

Section: The Locomotor Gaits Of Organismsmentioning

confidence: 99%

Synchrony and symmetry-breaking in active flagellar coordination

Wan

2019

Phil. Trans. R. Soc. B

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Living creatures exhibit a remarkable diversity of locomotion mechanisms, evolving structures specialised for interacting with their environment. In the vast majority of cases, locomotor behaviours such as flying, crawling, and running, are orchestrated by nervous systems. Surprisingly, microorganisms can enact analogous movement gaits for swimming using multiple, fast-moving cellular protrusions called cilia and flagella. Here, I demonstrate intermittency, reversible rhythmogenesis, and gait mechanosensitivity in algal flagella, to reveal the active nature of locomotor patterning. In addition to maintaining free-swimming gaits, I show that the algal flagellar apparatus functions as a central pattern generator which encodes the beating of each flagellum in a network in a distinguishable manner. The latter provides a novel symmetry-breaking mechanism for cell reorientation. These findings imply that the capacity to generate and coordinate complex locomotor patterns does not require neural circuitry but rather the minimal ingredients are present in simple unicellular organisms.

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Flagellum couples cell shape to motility inTrypanosoma brucei

Cited by 32 publications

References 57 publications

Electron cryo-tomography of vestibular hair-cell stereocilia

Electron cryo-tomography of vestibular hair-cell stereocilia

High-speed multifocal plane fluorescence microscopy for three-dimensional visualisation of beating flagella

Synchrony and symmetry-breaking in active flagellar coordination

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