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Clonal imaging of mitochondria in the dissected fly wing

Maddison, Daniel ORCID:, Mattedi, Francesca, Vagnoni, Alessio and Smith, Gaynor Ann ORCID: 2023. Clonal imaging of mitochondria in the dissected fly wing. Cold Spring Harbor Protocol 2023 (2) , 108051. 10.1101/pdb.prot108051

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Mitochondria are essential for long-term neuronal function and survival. They are maintained in neurons, including long axonal stretches, through dynamic processes such as fission, fusion, biogenesis and mitophagy. Here we describe a protocol for the in-depth morphological analysis of individual mitochondria in axons in vivo. Much of the mitochondrial analysis of axons is currently achieved in vitro where neurons are in a developmental state. Therefore, understanding the axonal mitochondrial network during aging in fully differentiated neurons and the long-term consequence of gene knockout is often missed. By using a clonal system paired with fluorescent genetically encoded markers in the Drosophila wing, we can visualize individual neurons (out of the whole bundle) including their long axons and the mitochondria that they contain using confocal imaging. The clonal system also allows visualization of neurons with genetic perturbations that would otherwise be lethal if present in the whole organism, allowing investigators to bypass lethality. This protocol can further be adapted to measure the physiological and biochemical state of the mitochondrion. Mitochondrial morphology and health in axons is tightly linked to aging, axon injury, and neurodegeneration; therefore, this method can be used to investigate the mitochondrial dysfunction associated with novel genes or those linked to neurodegenerative disease and axonopathy

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 20:48

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