<i>In situ</i> imaging of mitochondrial translation shows weak correlation with nucleoid DNA intensity and no suppression during mitosis

Estell, Christopher; Stamatidou, Emmanouela; El-Messeiry, Sarah; Hamilton, A. Ian

doi:10.1242/jcs.206714

Cited by 21 publications

(27 citation statements)

References 25 publications

(37 reference statements)

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“…To label newly synthesized mitochondrial‐encoded peptides, a click chemistry‐based approach was adapted (Estell et al , 2017). Briefly, cells were transferred to methionine‐free medium.…”

Section: Methodsmentioning

confidence: 99%

Monitoring mitochondrial translation in living cells

et al. 2021

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Mitochondria possess a small genome that codes for core subunits of the oxidative phosphorylation system and whose expression is essential for energy production. Information on the regulation and spatial organization of mitochondrial gene expression in the cellular context has been difficult to obtain. Here we devise an imaging approach to analyze mitochondrial translation within the context of single cells, by following the incorporation of clickable non‐canonical amino acids. We apply this method to multiple cell types, including specialized cells such as cardiomyocytes and neurons, and monitor with spatial resolution mitochondrial translation in axons and dendrites. We also show that translation imaging allows to monitor mitochondrial protein expression in patient fibroblasts. Approaching mitochondrial translation with click chemistry opens new avenues to understand how mitochondrial biogenesis is integrated into the cellular context and can be used to assess mitochondrial gene expression in mitochondrial diseases.

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Section: Methodsmentioning

confidence: 99%

Monitoring mitochondrial translation in living cells

et al. 2021

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“…However, a lack of appropriate imaging tools for mitochondrial translation has hindered assessing this subcellular event at single-cell levels. A recent study visualized mitochondrial translation using a non-canonical amino acid labeling in situ (Estell, Stamatidou, El-Messeiry, & Hamilton, 2017). This method allows the detection of mitochondrial translation at a singlecell resolution, but its application is limited to fixed cells.…”

Section: Discussionmentioning

confidence: 99%

Local protein synthesis of mtIF3 regulates mitochondrial translation for axonal development

Park

Lim

et al. 2021

Preprint

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Mitochondrial initiation factor 3 (mtIF3) binds to and dissociates mitochondrial ribosomes. The mtIF3-small subunit complex then recruits mtIF2, mRNA, and N-formylmethionine-tRNA to initiate mitochondrial translation. Intriguingly, transcripts of the nuclear-encoded mtIF3 gene have been shown present in axonal growth cones; however, the biological function of this compartmentalization remains largely unknown. Here, we demonstrate that brain-derived neurotrophic factor (BDNF) induces local translation of mtIF3 mRNA in axonal growth cones. Subsequently, mtIF3 protein is translocated into axonal mitochondria and promotes mitochondrial translation as assessed by our newly developed bimolecular fluorescence complementation sensor for the assembly of mitochondrial ribosomes. We further show that BDNF-induced axonal growth requires mtIF3-dependent mitochondrial translation in axons. These findings provide new insight into how neurons adaptively control mitochondrial physiology and axonal development via local mtIF3 translation.

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“…To label newly synthesized mitochondrial-encoded peptides, a click chemistry-based approach was adapted (Estell et al, 2017). Briefly, cells were transferred to methionine-free medium.…”

Section: Methodsmentioning

confidence: 99%

Local translation in synaptic mitochondria influences synaptic transmission

Yousefi

Fornasiero

Cyganek

et al. 2020

Preprint

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Mitochondria possess a small genome that codes for core subunits of the oxidative phosphorylation system, and whose expression is essential for energy production. Information on the regulation and spatial organization of mitochondrial gene expression in the cellular context has been difficult to obtain. Here we addressed this by devising an imaging approach to analyze mitochondrial translation, by following the incorporation of clickable non-canonical amino acids. We applied this method to multiple cell types, including hippocampal neurons, where we found ample evidence for mitochondrial translation in both dendrites and axons. Translation levels were surprisingly heterogeneous, were typically stronger in axons, and were independent of their distance from the cell soma, where mitochondria presumably descent from. Presynaptic mitochondrial translation correlated with local synaptic activity, and blocking mitochondria translation reduced synaptic function. Overall, these findings demonstrate that mitochondrial gene expression in neurons is intimately linked to neuronal function.

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In situ imaging of mitochondrial translation shows weak correlation with nucleoid DNA intensity and no suppression during mitosis

Cited by 21 publications

References 25 publications

Monitoring mitochondrial translation in living cells

Monitoring mitochondrial translation in living cells

Local protein synthesis of mtIF3 regulates mitochondrial translation for axonal development

Local translation in synaptic mitochondria influences synaptic transmission

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