Analysis of Mitochondrial Dynamics in Adult<i>Drosophila</i>Axons

Maddison, Daniel C.; Mattedi, Francesca; Vagnoni, Alessio; Smith, Gaynor A.

doi:10.1101/pdb.top107819

Cited by 3 publications

(1 citation statement)

References 88 publications

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“…It can thus be beneficial to age the fly between dissecting the first and second wings for an additional experimental time point. We have not observed any difference in results between flies that were aged with one wing dissected compared to flies that were aged with both wings with any of the markers listed in Table 1 of Introduction: Analysis of Mitochondrial Dynamics in Adult Drosophila Axons (Maddison et al 2022). Flies can also be used for mating purposes with one wing dissected.…”

Section: Methods Preparation Of Samplesmentioning

confidence: 82%

Clonal Imaging of Mitochondria in the Dissected Fly Wing

Maddison

Mattedi

Vagnoni

et al. 2022

Cold Spring Harb Protoc

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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. Most mitochondrial analysis of axons is currently performed in vitro with neurons in a developmental state. Therefore, an understanding of the axonal mitochondrial network during aging in fully differentiated neurons and the long-term consequence of gene knockout is often not developed. By using a clonal system paired with fluorescent genetically encoded markers in theDrosophilawing, 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 mitochondria. Mitochondrial morphology and health in axons are tightly linked to aging, axon injury, and neurodegeneration; therefore, this method can be used to investigate mitochondrial dysfunction associated with novel genes or those linked to neurodegenerative disease and axonopathy.

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Section: Methods Preparation Of Samplesmentioning

confidence: 82%

Clonal Imaging of Mitochondria in the Dissected Fly Wing

Maddison

Mattedi

Vagnoni

et al. 2022

Cold Spring Harb Protoc

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New insights into the functions of ACBD4/5-like proteins using a combined phylogenetic and experimental approach across model organisms

Kors,

Schuster,

Maddison

et al. 2024

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research

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New insights into the functions of ACBD4/5-like proteins using a combined phylogenetic and experimental approach across model organisms

Kors,

Schuster,

Maddison

et al. 2024

Preprint

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Acyl-CoA binding domain-containing proteins (ACBDs) perform diverse but often uncharacterised functions linked to cellular lipid metabolism. Human ACBD4 and ACBD5 are closely related peroxisomal membrane proteins, involved in tethering of peroxisomes to the ER and capturing fatty acids for peroxisomal β-oxidation. ACBD5 deficiency causes neurological abnormalities including ataxia and white matter disease. Peroxisome-ER contacts depend on an ACBD4/5-FFAT motif, which interacts with ER-resident VAP proteins. As ACBD4/5-like proteins are present in most fungi and all animals, we combined phylogenetic analyses with experimental approaches to improve understanding of their evolution and functions. Notably, all vertebrates exhibit gene sequences for both ACBD4 and ACBD5, while invertebrates and fungi possess only a single ACBD4/5-like protein. Our analyses revealed alterations in domain structure and FFAT sequences, which help understanding functional diversification of ACBD4/5-like proteins. We show that theDrosophila melanogasterACBD4/5-like protein possesses a functional FFAT motif to tether peroxisomes to the ER via Dm_Vap33. Depletion of Dm_Acbd4/5 caused peroxisome redistribution in wing neurons and reduced life expectancy. In contrast, the ACBD4/5-like protein of the filamentous fungusUstilago maydislacks a FFAT motif and does not interact with Um_Vap33. Loss of Um_Acbd4/5 resulted in an accumulation of peroxisomes and early endosomes at the hyphal tip. Moreover, lipid droplet numbers increased and mitochondrial membrane potential declined, implying altered lipid homeostasis. Our findings reveal differences between tethering and metabolic functions of ACBD4/5-like proteins across evolution, improving our understanding of ACBD4/5 function in health and disease. The need for a unifying nomenclature for ACBD proteins is discussed.

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Analysis of Mitochondrial Dynamics in AdultDrosophilaAxons

Cited by 3 publications

References 88 publications

Clonal Imaging of Mitochondria in the Dissected Fly Wing

Clonal Imaging of Mitochondria in the Dissected Fly Wing

New insights into the functions of ACBD4/5-like proteins using a combined phylogenetic and experimental approach across model organisms

New insights into the functions of ACBD4/5-like proteins using a combined phylogenetic and experimental approach across model organisms

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