iiinova — where your digital self keeps evolving

Shutt Lab

@mitomorph.bsky.social

1.2K followers 360 following 76 posts

Associate Professor at University of Calgary studying mitochondria.

Posts Replies Media

Shutt Lab @mitomorph.bsky.social · 03/06/2025

Congrats to ShuttLab PhD student Mashiat Zaman @mashiatzaman.bsky.social on receiving an award for his short talk at the recent CSMB-PRinCE meeting in Ottawa

070

Shutt Lab @mitomorph.bsky.social · 24/01/2025

Finally, through collaborations, we show that the reporter works in budding yeast (with Kurt Schmoller's lab) and in Drosophila (Mashiat again, with help from Abhi Sharma in Savraj Grewal's lab). *However, there are caveats associated with high expression, which can be detrimental.

Imaging of the Drosophila larval fat body. The mt-Kaede-HI-NESS reporter in the expressed from a UAS promoter driven by dau-Gal4. The Kaede mt-HI-NESS (green) is localized within the mitochondrial network (magenta). Nuclei are labelled with Hoechst (Cyan).

110

Shutt Lab @mitomorph.bsky.social · 24/01/2025

Mashiat also showed we can image extramitochondrial mtDNA and follow mtDNA in extracellular vesicles.

Live cell imaging of U2OS MFN2-KO or TFAM-Het lines known to have mtDNA release, which are transiently transfected with the mt-Kaede-HI-NESS reporter to visualize mtDNA nucleoids (green). Mitochondrial networks are labelled with Mitotracker DeepRed (purple). MtDNA nucleoids outside of the mitochondrial network are circled.

100

Shutt Lab @mitomorph.bsky.social · 24/01/2025

In the updated version, we show that we can use the reporter to quantify changes in mtDNA size and number (Mashiat Zaman in the Shutt lab). #FluorescenceFriday

Live cell imaging of U2OS cells (WT or MFN2 KO) that are either stably transduced or transiently transfected with the mt-Kaede-HI-NESS reporter (green) to visualize mtDNA nucleoids. Mitochondrial networks are labelled with Mitotracker DeepRed (purple).

130

Shutt Lab @mitomorph.bsky.social · 27/06/2024

Given the known links between mtDNA and inflammation, and the fact that inflammation can cause myopathy, we followed up in more detail, and found evidence for activation of both TLR9 and cGAS-STING inflammatory pathways.

100

Shutt Lab @mitomorph.bsky.social · 27/06/2024

When we characterized patient fibroblasts, we observed several phenotypes associated with MFN2 dysfunction. Excitingly, we also observed mtDNA puncta that were outside of the mitochondrial network and which co-localized with early endosomes.

100

Shutt Lab @mitomorph.bsky.social · 02/12/2023

For anyone attending ASCB who wants to learn more about our new mtDNA probe - Co-author and Collaborator Pina Colarusso will be presenting a poster Tuesday afternoon: www.biorxiv.org/content/10.1...

001

Shutt Lab @mitomorph.bsky.social · 30/10/2023

What was really great, was that we were able to photoactivate a subset of mtDNA nucleoids (magenta), which we could follow over time.

121

Shutt Lab @mitomorph.bsky.social · 30/10/2023

Next, we tried a different fluorescent protein, Kaede, which is photoswitchable from green to red. To our surprise, the mt-Kaede-HI-NESS worked beautifully, even with just a single H-NS DNA binding domain. (Mitos in grey and Kaede in cyan)

110

Shutt Lab @mitomorph.bsky.social · 30/10/2023

Excitingly, we were able to reduce the background mitochondrial network signal by adding a second H-NS DNA binding domain to get better nucleoid signal. Here you can see colocalization of TagRFP (magenta) with SYBR Gold labelled mtDNA nucleoids (cyan)

110

Shutt Lab @mitomorph.bsky.social · 30/10/2023

Our initial construct, mt-TagRFP-HI-NESS, worked OK. We could see nucleoid signal - Here you can see the TagRFP signal in puncta (magenta) within the mt network (grey) labelled with Mitotracker green. But there was also some background mitochondrial network signal in some cells.

121