Nanoresolution real-time 3D orbital tracking for studying mitochondrial trafficking in vertebrate axons in vivo
Abstract
We present the development and in vivo application of a feedback-based tracking microscope to follow individual mitochondria in sensory neurons of zebrafish larvae with nanometer precision and millisecond temporal resolution. By combining various technical improvements, we tracked individual mitochondria with unprecedented spatiotemporal resolution over distances of >100µm. Using these nanoscopic trajectory data, we discriminated five motional states: a fast and a slow directional motion state in both the anterograde and retrograde directions and a stationary state. The transition pattern revealed that mitochondria predominantly persist in the original direction of travel after a short pause, while transient changes of direction often exhibited longer pauses. Moreover, mitochondria in the vicinity of a second, stationary mitochondria displayed an increased probability to pause. The capability of following and optically manipulating a single organelle with high spatiotemporal resolution in a living organism offers a new approach to elucidating their function in its complete physiological context.
Data availability
The analysis software program is available on Gitlab and the wide-field images and trajectories are available on Zenodo. Source data files have been provided for all the figures.
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Wide-field Images and TrajectoriesZenodo, 10.5281/zenodo.2813946.
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Wide-field Images and TrajectoriesZenodo, 10.5281/zenodo.2815430.
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Wide-field Images and TrajectoriesZenodo, 10.5281/zenodo.281550.
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Wide-field Images and TrajectoriesZenodo, 10.5281/zenodo.2815703.
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Wide-field Images and TrajectoriesZenodo, 10.5281/zenodo.2815801.
Article and author information
Author details
Funding
Deutsche Forschungsgemeinschaft (SFB1032 (Project B3))
- Thomas Misgeld
- Don C Lamb
Fakultät für Chemie und Pharmazie, Ludwig-Maximilians-Universität München (Center for NanoScience (CeNS) and the BioImaging Network (BIN))
- Don C Lamb
H2020 European Research Council (ERC Grant Agreement n. 616791)
- Thomas Misgeld
German Center for Neurodegenerative Diseases
- Thomas Misgeld
Deutsche Forschungsgemeinschaft (research grants Mi 694/7)
- Thomas Misgeld
- Don C Lamb
Deutsche Forschungsgemeinschaft (Priority Program SPP1710)
- Thomas Misgeld
- Don C Lamb
Deutsche Forschungsgemeinschaft (SFB870 15 (Project A11))
- Thomas Misgeld
- Don C Lamb
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Thomas L Schwarz, Boston Children's Hospital, United States
Publication history
- Received: February 13, 2019
- Accepted: June 5, 2019
- Accepted Manuscript published: June 10, 2019 (version 1)
- Version of Record published: June 17, 2019 (version 2)
Copyright
© 2019, Wehnekamp et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Further reading
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- Physics of Living Systems
Schooling in fish is linked to a number of factors such as increased foraging success, predator avoidance, and social interactions. In addition, a prevailing hypothesis is that swimming in groups provides energetic benefits through hydrodynamic interactions. Thrust wakes are frequently occurring flow structures in fish schools as they are shed behind swimming fish. Despite increased flow speeds in these wakes, recent modeling work has suggested that swimming directly in-line behind an individual may lead to increased efficiency. However, only limited data are available on live fish interacting with thrust wakes. Here we designed a controlled experiment in which brook trout, Salvelinus fontinalis, interact with thrust wakes generated by a robotic mechanism that produces a fish-like wake. We show that trout swim in thrust wakes, reduce their tail-beat frequencies, and synchronize with the robotic flapping mechanism. Our flow and pressure field analysis revealed that the trout are interacting with oncoming vortices and that they exhibit reduced pressure drag at the head compared to swimming in isolation. Together, these experiments suggest that trout swim energetically more efficiently in thrust wakes and support the hypothesis that swimming in the wake of one another is an advantageous strategy to save energy in a school.
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- Physics of Living Systems
When a fish beats its tail, it produces vortices in the water that other fish could take advantage of to save energy while swimming.