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Inducing different severities of traumatic brain injury in Drosophila using a piezoelectric actuator

Nature Protocols volume 16, pages 263–282 (2021)Cite this article

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Abstract

Drosophila models have been instrumental in providing insights into molecular mechanisms of neurodegeneration, with wide application to human disease. The brain degeneration associated with traumatic brain injury (TBI) has been modeled in Drosophila using devices that inflict trauma on multiple parts of the fly body, including the head. However, the injuries produced by these models are not specific in location and are inconsistent between individual animals. We have recently developed a device that can be used to inflict controlled head injury to flies, resulting in physiological responses that are remarkably similar to those observed in humans with TBI. This protocol describes the construction, calibration and use of the Drosophila TBI (dTBI) device, a platform that employs a piezoelectric actuator to reproducibly deliver a force in order to briefly compress the fly head against a metal surface. The extent of head compression can be controlled through an electrical circuit, allowing the operator to set different levels of injury. The entire device can be assembled and calibrated in under a week. The device components and the necessary electrical tools are readily available and cost ~$800. The dTBI device can be used to harness the power of Drosophila genetics and perform large-scale genetic or pharmacological screens, using a 7-d post-injury survival curve to identify modifiers of injury.

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Fig. 1: Time-course of events after dTBI.
Fig. 2: Overview of experimental workflow for the dTBI paradigm.
Fig. 3: dTBI device overview.
Fig. 4: dTBI device wiring diagram.
Fig. 5: dTBI device setup and calibration.
Fig. 6: Representative results for low-throughput and high-throughput dTBI devices.

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Data availability

Source data are provided with this paper.

Code availability

The Arduino code used by the device is provided in the Supplementary Software 1.

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Acknowledgements

We thank M. Suplick and F. Letterio from the University of Pennsylvania Machine Shop for constructing and modifying the Heisenberg fly collars. We thank F. Carranza, A. Srinivasan and A. Perlegos for critical feedback. This work was supported by a predoctoral HHMI fellowship (J.S.), the Vagelos Molecular Life Sciences Scholars Program (J.K.), funding from the NIH R35-NS097275 (N.M.B.), the Paul G Allen Frontiers group and NIH NS088176 (D.F.M.).

Author information

Authors and Affiliations

  1. Department of Biology, University of Pennsylvania, Philadelphia, PA, USA

    Janani Saikumar, Joshua Kim & Nancy M. Bonini

  2. Neuroscience Graduate Group, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    China N. Byrns & Nancy M. Bonini

  3. Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA

    Matthew Hemphill & David F. Meaney

  4. Department of Neurosurgery, University of Pennsylvania, Philadelphia, PA, USA

    David F. Meaney

Authors
  1. Janani Saikumar
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  2. Joshua Kim
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  3. China N. Byrns
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  4. Matthew Hemphill
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  5. David F. Meaney
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  6. Nancy M. Bonini
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Contributions

All experiments were performed by J.S. and J.K. under the mentorship of N.M.B. The piezoelectric dTBI device was designed by D.F.M., M.H. and J.S.; M.H. and D.F.M. built the device, and J.S., J.K. and C.N.B. tested it. J.S., D.F.M. and N.M.B. wrote the manuscript, and J.K. and C.N.B. provided critical feedback.

Corresponding authors

Correspondence to David F. Meaney or Nancy M. Bonini.

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Competing interests

The authors declare no competing interests.

Additional information

Peer review information Nature Protocols thanks Douglas M. Ruden and David Wassarman for their contribution to the peer review of this work.

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Related links

Key reference using this protocol

Saikumar, J. et al. Proc. Natl Acad. Sci. USA 117, 17269–17277 (2020): https://www.pnas.org/content/117/29/17269

Supplementary information

Supplementary Information

Supplementary Videos 1–4 legends, Supplementary Software 1 legend, Supplementary Figs. 1–6 and legends.

Reporting Summary

Supplementary Video 1

Severe compression in the low-throughput and high-throughput devices.

Supplementary Video 2

Calibration and compression measurements in the low-throughput device: The measurements of % head compression, the gap between the collar surface and piezoelectric actuator (purple), height of uncompressed fly head (green), height of fly head at the point of maximum compression (blue), and the gap between the head and the piezoelectric actuator (red) corresponding in location to the schematic in Fig 5b.

Supplementary Video 3

Collar loading and unloading protocol: A step-by-step guide on using a pair of blunt forceps to load the fly into the collar and move it along the groove with the forceps or a paintbrush.

Supplementary Video 4

dTBI procedure: A step-by-step guide on positioning the collar under the piezoelectric actuator and performing the injury.

Supplementary Software 1

Arduino code to operate dTBI device.

Source data

Source Data Fig. 6

Raw data for regression analyses in Fig 6 a, b and lifespan analyses in Fig. 6 c, d.

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Saikumar, J., Kim, J., Byrns, C.N. et al. Inducing different severities of traumatic brain injury in Drosophila using a piezoelectric actuator. Nat Protoc 16, 263–282 (2021). https://doi.org/10.1038/s41596-020-00415-y

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