Live imaging of mitochondria in the intact fly wing

Mattedi, Francesca, Maddison, Daniel ORCID: https://orcid.org/0000-0003-3038-1687, Smith, Gaynor Ann ORCID: https://orcid.org/0000-0003-4332-8383 and Vagnoni, Alessio 2023. Live imaging of mitochondria in the intact fly wing. Cold Spring Harbor Protocol 2023 (2) , 108052. 10.1101/pdb.prot108052

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Abstract

Detailed mechanisms governing the transport of mitochondria in neurons have recently emerged, although it is still poorly understood how the regulation of transport is coordinated in space and time within the physiological context of an organism. Here, we provide a protocol to study the intracellular dynamics of mitochondria in the wing neurons of adult Drosophila in situ. The mounting and imaging procedures that we describe are suitable for use on most microscopes, and they can be easily implemented in any laboratory. Our noninvasive mounting procedures, combined with the translucency of the wing cuticle in adult animals, makes the wing nervous system accessible to advanced microscopy studies in a physiological environment. Combining the powerful genetics of Drosophila with time-lapse live imaging, users of this protocol will be able to analyze mitochondrial dynamics over time in a subset of sensory neurons in the wing. These cells extend long axons with a stereotypical plus-end-out microtubule orientation that represents a unique model to understand the logic of neuronal cargo transport, including the mitochondria. Finally, the neurons in this tissue respond to mechanical and chemical stimulation of the sensory organs of the wing, opening up the possibility of coupling the study of mitochondrial dynamics with the modulation of neuronal activity in aging Drosophila. We anticipate that the unique characteristics of this in vivo system will contribute to the discovery of novel mechanisms that regulate mitochondrial dynamics within an organismal context with relevant implications for the pathogenesis of age-dependent neurological disorders.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine MRC Centre for Neuropsychiatric Genetics and Genomics (CNGG)
Publisher:	Cold Spring Harbor Laboratory Press
ISSN:	1940-3402
Date of First Compliant Deposit:	8 November 2022
Date of Acceptance:	30 September 2022
Last Modified:	06 Jan 2024 15:56
URI:	https://orca.cardiff.ac.uk/id/eprint/154037

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